RNA interference(RNAi)is a post-transcriptional gene-silencing technique induced by the introduction of double-stranded RNA(dsRNA)or small interfering RNA(siRNA)[1].RNAi-based strategies have been extensively applied ...RNA interference(RNAi)is a post-transcriptional gene-silencing technique induced by the introduction of double-stranded RNA(dsRNA)or small interfering RNA(siRNA)[1].RNAi-based strategies have been extensively applied in the treatment of human diseases and crop protection against insect pests[2-4].With the availability of the full genome sequences of major mosquito vectors,RNAi has become increasingly used as a novel means of mosquito control[5].展开更多
RNA interference(RNAi)has been used for agricultural insect pest control based on silencing of targeted insect genes.However,the effectiveness of RNAi and its applications in insect pest control remain challenging.Her...RNA interference(RNAi)has been used for agricultural insect pest control based on silencing of targeted insect genes.However,the effectiveness of RNAi and its applications in insect pest control remain challenging.Here we review factors that may affect the effectiveness of RNAi application,including the variability in RNAi efficacy among different insect species,a limited understanding of double-stranded RNA(dsRNA)uptake and systemic RNAi mechanisms,and the effective delivery of dsRNA in field conditions.Furthermore,we summarize recent progress in RNAi strategies for crop protection,discuss the advantages and disadvantages of RNAi-based insect control,and propose potential strategies to increase the effectiveness of RNAi in insect control.展开更多
Nucleic acid drugs represent the third wave of innovation in drug research and development,succeeding small-molecule and antibody drugs.These drugs,particularly RNA interference(RNAi)therapies,have become a pivotal fo...Nucleic acid drugs represent the third wave of innovation in drug research and development,succeeding small-molecule and antibody drugs.These drugs,particularly RNA interference(RNAi)therapies,have become a pivotal focus in the pharmaceutical industry.RNAi drugs are extensively utilized in the treatment of chronic and rare diseases due to their exceptional gene-silencing efficiency,manageable side effects,and straightforward synthesis process.This study undertook a thorough analysis of the global landscape of RNAi drug patents,highlighting the latest technological advancements and trends.We meticulously identified and cataloged the key technologies that dominated this patent landscape.The goal was to provide valuable insights and references for researchers involved in the development of RNAi drugs within the domestic pharmaceutical sector.展开更多
FOXL 2 and CYP 19 B are crucial transcription factors in vertebrates and invertebrates that play pivotal roles in sex differentiation and gonadal development.The potential roles of the foxl 2 and cyp 19 b genes in sex...FOXL 2 and CYP 19 B are crucial transcription factors in vertebrates and invertebrates that play pivotal roles in sex differentiation and gonadal development.The potential roles of the foxl 2 and cyp 19 b genes in sex determination and gonadal development in Cyprinus carpio var.koi were explored using a non-invasive RNA interference(RNAi)method,histopathological observation and qPCR.Results demonstrate that foxl 2 exhibited a sexually dimorphic expression pattern in gonads,with a notable expression in ovaries;cyp 19 b was expressed in all peripheral tissues,with a particularly prominent expression in brain and gonads.The knockdown of foxl 2 by RNAi resulted in delay in the development of the female gonads.Conversely,no notable alterations were discerned in the gonads of C.carpio var.koi following the knockdown of cyp 19 b.The upregulation of sox 9 a,amh,and cyp 19 b following foxl 2 knockdown indicates that foxl 2 may play a pivotal role in gonadal development.Nevertheless,further investigation is required to ascertain the potential role of cyp 19 b.This study elucidated the role of foxl 2 and enhanced the understanding of the mechanisms of sex determination and gonadal development in C.carpio var.koi.展开更多
The means of survival of genomic dsRNA of reoviruses from dsRNA-triggered and Dicer-initiated RNAi pathway remains to be defined. The present study aimed to investigate the effect of Grass carp reovirus (GCRV) replica...The means of survival of genomic dsRNA of reoviruses from dsRNA-triggered and Dicer-initiated RNAi pathway remains to be defined. The present study aimed to investigate the effect of Grass carp reovirus (GCRV) replication on the RNAi pathway of grass carp kidney cells (CIK). The dsRNA-triggered RNAi pathway was demonstrated unimpaired in CIK cells through RNAi assay. GCRV-specific siRNA was generated in CIK cells transfected with purified GCRV genomic dsRNA in Northern blot analysis; while in GCRV-infected CIK cells, no GCRV-specific siRNA could be detected. Infection and transfection experiments further indicated that replication of GCRV correlated with the increased transcription level of the Dicer gene and functional inhibition of in vitro synthesized egfp-siRNA in silencing the EGFP reporter gene. These data demonstrated that although only the genomic dsRNA of GCRV was sensitive to the cellular RNAi pathway, unidentified RNAi suppressor protein(s) might contribute to the survival of the viral genome and efficient viral replication.展开更多
Objective: The effects on cell-cycle and p53 expression in hepatoma cell line SK-Hep-1 were explored by transfecting exogenous p53 small double stranded RNA (dsRNA) into the SK-Hep-1 cells. Methods: p53 dsRNA and EGFP...Objective: The effects on cell-cycle and p53 expression in hepatoma cell line SK-Hep-1 were explored by transfecting exogenous p53 small double stranded RNA (dsRNA) into the SK-Hep-1 cells. Methods: p53 dsRNA and EGFP dsRNA were synthesized. SK-Hep-1 (wtp53) cell line was transfected with 200 ng and 400 ng p53 dsRNA or EGFP and EGFP+EGFP dsRNA (as positive control) or 9% NaCl (as blank control) by liposome transfection technique. Flow cytometry was adopted to measure the effects of p53 dsRNA on cell cycle. Expression of p53 protein was detected by Western-Blotting at 48 h after transfecting p53 dsRNA. Results: The number of G0-G1 phase SK-Hep-1 cells, which were transfected with 200 ng p53 dsRNA, was decreased by 52.53% comparing with the control, and decreased by 50.29% (P<0.05) comparing with the positive control cells transfected with same dosage of EGFP+EGFP dsRNA. The number of S phase cells, which were transfected with 200 ng p53 dsRNA, was increased by 146.8% comparing with the control, and increased by 128.62% (P<0.05) comparing with the positive control cells transfected with same dosage of EGFP+EGFP dsRNA. The number of G2-M phase cells, which were transfected with 200 ng p53 dsRNA, was increased by 30.56% (P<0.05) comparing with the control, and increased by 21.63% (P>0.05) comparing with the positive control cells transfected with same dosage of EGFP+EGFP dsRNA. After 48 h, p53 protein expression was not detected in the SK-Hep-1 cells transfected with p53 dsRNA. Conclusion: p53 dsRNA can obviously improve the proliferation of SK-Hep-1 cells, and suppress p53 protein expression of SK-Hep-1 cells, the former may be related to of the latter.展开更多
Objective： Lung cancer has emerged as a leading cause of cancer death in the world. Current therapies are ineffective, thus new approaches are needed to improve the therapeutic ratio. RNA interference （RNAi） has sh...Objective： Lung cancer has emerged as a leading cause of cancer death in the world. Current therapies are ineffective, thus new approaches are needed to improve the therapeutic ratio. RNA interference （RNAi） has shown promise in gene silencing in vitro, the potential of which in developing new methods for the therapy of non-small-cell lung cancer （NSCLC） needs to be further tested in vivo. In this study, chemically synthesized double-stranded RNA （dsRNA） targeting epidermal growth factor receptor （EGFR） was transfected into NSCLC cell line SPC-A1 cells and established the tumor burdened athymic nude mice model to investigate whether dsRNA could induce gene silencing in NSCLC cells in vivo. Methods： SPC-A1 was transfected with EGFR sequence-specific dsRNA formulated with Lipofectamine 2000. SPC-A1 cells （1 × 107/ mL） in 200 pL were injected s.c. into the left flank area of the mice to establish the tumor burdened athymic nude mice model. Calculate the tumor growth inhibition rate by measuring the diameter and the weight of the tumor. Immunohistochemistry and Westem blot were used to monitor the reduction in the production of the EGFR protein. Realtime RT-PCR was used to detect the silencing of the EGFR mRNA level. Results： It displayed that EGFR sequence specific dsRNA （dsRNA-EGFR） significantly inhibited the tumor growth in vivo. The tumor growth inhibition rate was 75.03%. The dsRNA-EGFR sequence specifically silenced EGFR with 53.6% of down-regulation of EGFR protein production and 32.3% of silencing of EGFR mRNA level. Conclusion： DsRNA-EGFR showed a blockbuster effect in downregulation of EGFR mRNA level and protein production, and inhibition of tumor growth in vivo.展开更多
Tobacco(Nicotiana tabacum)is a widely used platform for producing recombinant proteins for clinical applications.However,achieving mammalian-like glycosylation modifications in plantderived therapeutic proteins remain...Tobacco(Nicotiana tabacum)is a widely used platform for producing recombinant proteins for clinical applications.However,achieving mammalian-like glycosylation modifications in plantderived therapeutic proteins remains challenging,particularly in the case of fucosylation mediated by fucosyltransferases(FUTs).In this study,an RNA interference(RNAi)plasmid targeting the first exon ofα-1,3-fucosyltransferase 4(FUT4)gene was constructed,named as FUT4-RNAi.Using Agrobacterium-mediated transformation with EHA105 harboring the FUT4-RNAi plasmid,we obtained 29 regenerated tobacco lines,17 of which were confirmed as putatively positive by PCR.The mRNA transcript accumulation of the FUT4 gene was significantly reduced in 16 out of the 17 transgenic lines compared to the negative control,cv.Yunyan 87.Among these,11 lines(4^(#),6,7,11^(#),12^(#),15^(#),19^(#),22^(#),26^(#),28^(#) and 29^(#))showed FUT4 transcript levels below 25%of those in cv.Yunyan 87.Four lines(7^(#),12^(#),15^(#),and 29^(#))with the lowest mRNA levels were selected for further analysis by western blotting(WB)and enzyme-linked immunosorbent assay(ELISA).The results confirmed a significant decrease in FUT4 protein levels in these lines compared with that in cv.Yunyan 87,with line 29*showing less than 13%of the FUT4 protein content compared to cv.Yunyan 87.This study successfully developed a tobacco chassis with severely downregulated FUT4 expression,laying an important foundation for the production of human therapeutic proteins using a plant expression system.展开更多
Bone morphogenetic proteins(BMPs) play a critical role in the growth and steroidogenesis of granulosa cells(GCs).BMP signals act through membrane-bound heteromeric serine/threonine kinase receptors.Upon ligand binding...Bone morphogenetic proteins(BMPs) play a critical role in the growth and steroidogenesis of granulosa cells(GCs).BMP signals act through membrane-bound heteromeric serine/threonine kinase receptors.Upon ligand binding,BMPs activate intracellular Smad proteins and regulate growth and apoptosis in various cell types.The objective of this study was to demonstrate the effects of BMP/Smad signal on growth and steroidogenesis of porcine GCs.A strategy of RNA interference(RNAi)-mediated 'gene silencing' of Smad4,a core molecule mediating the intracellular BMP/Smad signal transduction pathways,was used to interrupt endogenous BMP/Smad signaling.Results indicate that Smad4-small interfering RNA(siRNA) caused specific inhibition of Smad4 mRNA and protein expression after transfection.Interrupted endogenous BMP/Smad signaling significantly inhibited growth,and induced apoptosis of porcine GCs,while decreasing estradiol production.In addition,interrupted BMP/Smad signaling significantly(P<0.05) changed the expression of Cyclin D2,CDK4,Bcl-2,and Cyp19a1.These findings provide new insights into how BMP/Smad signaling regulates the growth and steroidogenesis of porcine GCs.展开更多
Modern agribusiness plays a vital role in safeguarding and improving the production,quality,and quantity of food,feed,fiber,and fuel.Growing concerns over the impact of chemical pesticides on health and the environmen...Modern agribusiness plays a vital role in safeguarding and improving the production,quality,and quantity of food,feed,fiber,and fuel.Growing concerns over the impact of chemical pesticides on health and the environment have stimulated the industry to search for alternative and greener solutions.Over the last years,the RNA interference(RNAi)process has been identified as a very promising new approach to complement the arsenal of foliar spray,soil,or seed treatments applied as chemical and biological pest control agents,and of plant-incorporated protectants(PIPs).RNA-based active ingredients(AIs)possess a unique mode of action and can be implemented via both genetic modification(GM)and biocontrol approaches.RNA-based AIs promise to deliver the selectivity and sustainability desired in future crop protection agents.This is due to their utilization of a natural process to exert control and their high level of selectivity,which leads to reduced risk for non-target organisms(NTOs).This review discusses the advantages and limitations of RNA-based solutions in crop protection and recent research progress toward RNA-based biocontrols against the Colorado potato beetle(CPB),corn rootworm(CRW),and soy stink bug(SSB).Many challenges still exist on the road to the implementation of a broad range of RNA-based products and their widespread use and application.Despite these challenges,it can be expected that RNA-based AIs will become valuable new tools complementing the current arsenal of crop-protection solutions.展开更多
OBJECTIVE To explore the role of gecko crude peptides(GCPs)in the proliferation,apoptosis,migration and lymphangiogenesis of human hepatocellular carcinoma cells(Hep G2)and human lymphaticendothelial cells(HLECs)in vi...OBJECTIVE To explore the role of gecko crude peptides(GCPs)in the proliferation,apoptosis,migration and lymphangiogenesis of human hepatocellular carcinoma cells(Hep G2)and human lymphaticendothelial cells(HLECs)in vitro.METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was used to evaluate the anti-proliferative effect of GCPs and si RNA-VEGF-C on Hep G2 cells,Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis.The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay.The protein and m RNA expressions of vascular endothelial growth factor-C(VEGF-C)and CXC chemokine receptor-4(CXCR4)were detected by q-PCR,immunofluorescence,Western blot.The protein expressions of the extracellular signal regulated kinase(ERKI/2),c-Jun N-terminal kinase(JNK),p38-mitogen activated protein kinases(p38 MAPK),serine/threonine kinase(Akt)and phosphatidylinositol-3-kinase(PI3K)were detected by western blot.The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay.The protein and m RNA expressions of vascular endothelial growth factor receptor-3(VEGFR-3)and stromal cell-derived factor-1(SDF-1)were detected by q-PCR,Western blot.RESULTS GCPs and si RNA-VEGF-C inhibited Hep G2 proliferation,invasion and migration,and the most obvious inhibitory effect was both synergistic effects.Thus,GCPs suppressed HLECs proliferation,migration and tubelike structure formationin a dose-dependent manner,and had inhibitory effect of tumor-induced lymphangiogenesis in vitro.Additionally,we found that GCPs and si RNA-VEGF-C decreased the expressions of MMP-2,MMP-9,VEGF-C,CXCR4,phospho-ERK1/2,phospho-P38,phospho-JNK and PI3K in Hep G2 cells.Moreover,GCPs had a dose-dependent depressive effecton the expressions of VEGFR-3,SDF-1 in HLECs.CONCLUSION The low expression of VEGF-C mediated by si RNA-VEGF-C and GCPs inhibit tumor proliferation,invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C,and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.展开更多
Objective： To investigate the inhibitory effects of RNAi （ RNA interference, RNAi） expression vector on CXCR4 expression in prostate carcinoma cell lines. Methods： Small interference RNA （siRNA） expression vecto...Objective： To investigate the inhibitory effects of RNAi （ RNA interference, RNAi） expression vector on CXCR4 expression in prostate carcinoma cell lines. Methods： Small interference RNA （siRNA） expression vectors for CXCR4 gene were constructed and transfected into prostate carcinoma cell lines（PC-3m and LNCaP）with liposomes. T expression of CXCR4 was detected by RT-PCR and western blot. Results： T expression of CXCR4 mRNA and protein in the PC-3m and LNCaP cells was reduced by RNAi expression vectors. The inhibitory rate of CXCR4 mRNA expression in the PC-3m cells was 87.81% ± 10.20% ,56.10% ± 9.32% at the 24th hour and the 48th hour, compared with 56.93% ±8.78% ,49.24% ± 11.23% in LNCaP cells. The inhibitory rate of the expression of CXCR4 protein was 64.71% ± 6.68% ,58.66% ± 11.56% respectively. Conclusion： The expression of CXCR4 gene can effectively be inhibited by RNAi expression vectors.展开更多
High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression oc...High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression occurrs in many kinds of cancers including colorectal cancer,suggesting that HMGA2 might play a critical role in the progression of various tumors.However,the exact role of HMGA2 in colorectal cancer has not been determined.To verify the essential role of HMGA2 in the growth and invasiveness of colorectal cancer,HMGA2 expression was down-regulated by RNA interference（RNAi） in SW480 cells.We observed that the knockdown of HMGA2 led to the significant inhibition of proliferation and invasion of SW480 cells in vitro.These results suggest that HMGA2 might play a crucial role in the progression of colorectal cancer,and be a potential therapeutic target for human colorectal cancer.展开更多
Objective：To investigate the silencing effects of recombinant adenovirus Ad-shRNA-MK on midkine（MK） gene in pancreatic cancer cells. Methods：Ad-shRNA-MK was used to infect pancreatic cancer BxPC-3 cells. Assays we...Objective：To investigate the silencing effects of recombinant adenovirus Ad-shRNA-MK on midkine（MK） gene in pancreatic cancer cells. Methods：Ad-shRNA-MK was used to infect pancreatic cancer BxPC-3 cells. Assays were conducted for knockdown of the MK gene on the day of infection and on the 1 ^st, 3^rd, 5^th, 7^th, and 9^th days post-infection by using immunocytochemistry, real-time RT-PCR, and Western blot analysis. Results：The adenoviral Ad-shRNA-PTN was constructed successfully, and infection was confirmed by electron microscopic observation. By using real-time RT-PCR, the inhibition rates of MK mRNA expression in the BxPC-3 cells were 20%, 80%, 55%, and 23% on the 1st, 3^th, 5^th, and 7^th days post-infection. Immunocytochemistry and Western blot analysis confirmed this effect at the gene product level. Conclusion：Efficient and specific knockdown of MK in pancreatic cancer cells by adenoviral Ad-shRNA-PTN is a potentially powerful tool for the study of gene therapy of pancreatic cancer nerve infiltration.展开更多
Objectives:In this study,we explored how adiponectin mediated urotensinⅡ(UⅡ)-induced tumor necrosis factor-α(TNF-α)andα-smooth muscle actin(α-SMA)expression and ensuing intracellular signaling pathways in advent...Objectives:In this study,we explored how adiponectin mediated urotensinⅡ(UⅡ)-induced tumor necrosis factor-α(TNF-α)andα-smooth muscle actin(α-SMA)expression and ensuing intracellular signaling pathways in adventitial fibroblasts(AFs).Methods:Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UⅡand inhibitors of signal transduction pathways for 1-24 h.The cells were then harvested for TNF-αreceptor(TNF-α-R)messenger RNA(mRNA)and TNF-αprotein expression determination by reverse transcription-polymerase chain reaction(RT-PCR)and enzyme-linked immunosorbent assay(ELISA),respectively.Adiponectin and adiponectin receptor(adipoR)expression was measured by RT-PCR,quantitative real-time PCR(qPCR),immunohistochemical analysis,and cell counting kit-8(CCK-8)cell proliferation experiments.We then quantified TNF-αandα-SMA mRNA and protein expression levels by qPCR and immunofluorescence(IF)staining.RNA interference(RNAi)was used to explore the function of the adipoR genes.To investigate the signaling pathway,we applied western blotting(WB)to examine phosphorylation of adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK).In vivo,an adiponectin(APN)-knockout(APN-KO)mouse model mimicking adventitial inflammation was generated to measure TNF-αandα-SMA expression by application of qPCR and IF,with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling.Results:In both cells and tissues,UⅡpromoted TNF-αprotein and TNF-α-R secretion in a dose-and time-dependent manner via Rho/protein kinase C(PKC)pathway.We detected marked expression of adipoR1,T-cadherin,and calreticulin as well as a moderate presence of adipoR2 in AFs,while no adiponectin was observed.Globular adiponectin(gAd)fostered the growth of AFs,and acted in concert with UⅡto induceα-SMA and TNF-αthrough the adipoR1/T-cadherin/calreticulin/AMPK pathway.In AFs,gAd and UⅡsynergistically induced AMPK phosphorylation.In the adventitial inflammation model,APN deficiency up-regulated the expression ofα-SMA,UⅡreceptor(UT),and UⅡwhile inhibiting TNF-αexpression.Conclusions:From the results of our study,we can speculate that UⅡinduces TNF-αprotein and TNF-α-R secretion in AFs and rat tunica adventitia of vessels via the Rho and PKC signal transduction pathways.Thus,it is plausible that adiponectin is a major player in adventitial progression and could serve as a novel therapeutic target for cardiovascular disease administration.展开更多
Objective:Survivin has attracted abundant interest in tumor research since it was discovered in 1997.But several studies indicated that the relationship between survivin expression and tumor behavior is still not full...Objective:Survivin has attracted abundant interest in tumor research since it was discovered in 1997.But several studies indicated that the relationship between survivin expression and tumor behavior is still not fully understood.So our main aim was to observe the localization of survivin in colon cancer and its influence on the colon cancer biocharacteristics.Methods:We screened the expression and localization of survivin in SW480 by immunofluorescence and immunocytochem-istry.Then, we constructed the recombinant adenovirus (Ad-survivin/shRNA), which contained the short hairpin RNA (shRNA) of survivin and transfected it into SW480 cells.We detected survivin gene expression after shRNA interference by RT-PCR and Western blot, and its influence on the proliferation, apoptosis and cell cycle was analyzed by MTT, colony-formation, and flow cytometry.Results:Survivin was expressed at a high level in SW480 cells and the sub-cellular localization was in the cytoplasm.Recombinant adenovirus could suppress survivin expression efficiency and inhibit proliferation, induce apoptosis, and affected the mitosis in vitro.Conclusion:Survivin plays an important role in controlling tumor growth by a variety of molecular regulatory mechanisms.Cytoplasmic survivin silence could induce apoptosis, effect the mitotic cycle and inhibit cell proliferation.展开更多
Objective: Genome-wide association studies(GWAS) have identified over 150 risk loci linked to colorectal cancer(CRC), including the 17p13.3 locus with the tag single nucleotide polymorphism(SNP) rs12603526 in the Asia...Objective: Genome-wide association studies(GWAS) have identified over 150 risk loci linked to colorectal cancer(CRC), including the 17p13.3 locus with the tag single nucleotide polymorphism(SNP) rs12603526 in the Asian population. However, the specific causal gene and the functional regulatory mechanisms in this region remain unresolved, necessitating further investigation to elucidate the underlying mechanisms of CRC.Methods: We employed an RNA interference-based functional approach to identify genes critical for CRC cell proliferation at the GWAS locus 17p13.3. Bioinformatic fine-mapping analysis was conducted to prioritize causal variants. A large-scale study involving 7,013 cases and 7,329 controls from a Chinese population, along with another cohort of 5,158 cases and 20,632 controls from the UK Biobank, was performed to validate the association between the candidate variant and the gene. A series of biological experiments was conducted to explore the function of the candidate gene and its regulatory mechanisms.Results: We identified FAM57A as a key oncogene that promotes CRC cell proliferation, and confirmed its carcinogenic role through in vitro proliferation assays. The variant rs526835 was prioritized as a causal candidate for CRC risk, located in a functional region with enhancer properties, and showed a significant quantitative association with FAM57A expression. The rs526835 [T] variant was associated with a 1.17-fold increase in CRC risk [95%confidence interval(95% CI): 1.11-1.23, P=1.23×10^(−9)] in the large-scale Chinese cohort, which was further corroborated in the UK Biobank cohort. Mechanistically, we demonstrated that rs526835 enhances a promoterenhancer interaction mediated by the transcription factor JUN, leading to increased expression of FAM57A.Conclusions: We reveal the underlying mechanisms of CRC predisposition at the GWAS locus 17p13.3.Additionally, our findings highlight the critical role of FAM57A in CRC pathogenesis and introduce a novel enhancer-promoter interaction between FAM57A and rs526835, which could inform future precision prevention and personalized cancer therapies.展开更多
Objective:The aim of this study was to study the changes of SGC-7901 cells transfecting small hairpin RNA(shRNA) targeted Bcl-2 and celecoxib in vitro.Methods:To use the effective siRNA targeted to Bcl-2 by Lipofectam...Objective:The aim of this study was to study the changes of SGC-7901 cells transfecting small hairpin RNA(shRNA) targeted Bcl-2 and celecoxib in vitro.Methods:To use the effective siRNA targeted to Bcl-2 by Lipofectamine 2000,the rate of cell growth inhibition of all groups was detected by multiply-table tournament(MTT) method,apoptosis rate was examined by flow cytometry and the expression of Bcl-2 was assayed by Elisa.Results:After siRNA was transfected to SGC-7901 cells,the rate of cell growth inhibition was increased,the joint group was higher by flow cytometry and the expression of Bcl-2 was lower by Elisa.Conclusion:The growth of SGC-7901 cells which was transfected siRNA has been inhibited,and the sensitivity to celecoxib has been increased.展开更多
The paper aimed to study the effect of lysyl oxidase-like 2 （LOXL2） on hepatocellular carcinoma （HCC） and explore the biological mechanisms of tumorigenicity and progression in HCC. The authors used four HCC cell ...The paper aimed to study the effect of lysyl oxidase-like 2 （LOXL2） on hepatocellular carcinoma （HCC） and explore the biological mechanisms of tumorigenicity and progression in HCC. The authors used four HCC cell lines to identify LOXL2. A lentiviral vector containing LOXL2-siRNA was constructed to silence the LOXL2 gene in SMMC-7721 cell line, and mRNA of the target gene was detected by real-time polymerase chain reaction （RT-PCR）. The effect of LOXL2 silencing on the growth of SMMC-7721 cells was explored with flow cytometry profiling and BrdU labeling. Downstream genes of LOXL2 were selected by microarray and verified by Western Blotting. In the results, LOXL2 expression was significantly up-regulated in four types of HCC cell lines, therefore, SMMC-7721 cell line was selected for further exploration. When SMMC-7721 cell line was infected with LOXL2-siRNA, the expression of LOXL2 mRNA decreased. The silencing of LOXL2 resulted in the cell cycle arrest at the G 1-phase, the increased apoptosis and the decreased growth of SMMC-7721 cells on the indicated days by BrdU. Moreover, the MDM2, BIRC3, CDC42, FOS and TGFBR2 genes were selected and verified to be the downstream genes of LOXL2. In conclusion, LOXL2 contributes to the genesis and progression of HCC cells and works by regulating downstream genes of LOXL2 in certain pathways. Therefore, LOXL2 may play an important role in the progression and prognosis of HCC.展开更多
基金supported by grants from the National Key Research and Development Program(2023YFE0113600).
文摘RNA interference(RNAi)is a post-transcriptional gene-silencing technique induced by the introduction of double-stranded RNA(dsRNA)or small interfering RNA(siRNA)[1].RNAi-based strategies have been extensively applied in the treatment of human diseases and crop protection against insect pests[2-4].With the availability of the full genome sequences of major mosquito vectors,RNAi has become increasingly used as a novel means of mosquito control[5].
基金funded by the National Natural Science Foundation of China(32188102 to Lanqin Xia)Natural Science Foundation for Distinguished Young Scholars of Jiangxi province(20212ACB215001 to Xiudao Yu)+1 种基金supported by the China Scholarship Council(202303250062)the GSCAAS-ULg Joint PhD Program。
文摘RNA interference(RNAi)has been used for agricultural insect pest control based on silencing of targeted insect genes.However,the effectiveness of RNAi and its applications in insect pest control remain challenging.Here we review factors that may affect the effectiveness of RNAi application,including the variability in RNAi efficacy among different insect species,a limited understanding of double-stranded RNA(dsRNA)uptake and systemic RNAi mechanisms,and the effective delivery of dsRNA in field conditions.Furthermore,we summarize recent progress in RNAi strategies for crop protection,discuss the advantages and disadvantages of RNAi-based insect control,and propose potential strategies to increase the effectiveness of RNAi in insect control.
文摘Nucleic acid drugs represent the third wave of innovation in drug research and development,succeeding small-molecule and antibody drugs.These drugs,particularly RNA interference(RNAi)therapies,have become a pivotal focus in the pharmaceutical industry.RNAi drugs are extensively utilized in the treatment of chronic and rare diseases due to their exceptional gene-silencing efficiency,manageable side effects,and straightforward synthesis process.This study undertook a thorough analysis of the global landscape of RNAi drug patents,highlighting the latest technological advancements and trends.We meticulously identified and cataloged the key technologies that dominated this patent landscape.The goal was to provide valuable insights and references for researchers involved in the development of RNAi drugs within the domestic pharmaceutical sector.
基金Supported by the Qingdao Aquarium Technology Collaborative Innovation Center Cooperation Project(No.20210021)the Researching Key Technologies for Selecting Excellent Koi Carp Germplasm(No.20223702032291)the Qingdao Agricultural University Tangwang Koi Carp Joint R&D Center Collaborative Project(No.20220271)。
文摘FOXL 2 and CYP 19 B are crucial transcription factors in vertebrates and invertebrates that play pivotal roles in sex differentiation and gonadal development.The potential roles of the foxl 2 and cyp 19 b genes in sex determination and gonadal development in Cyprinus carpio var.koi were explored using a non-invasive RNA interference(RNAi)method,histopathological observation and qPCR.Results demonstrate that foxl 2 exhibited a sexually dimorphic expression pattern in gonads,with a notable expression in ovaries;cyp 19 b was expressed in all peripheral tissues,with a particularly prominent expression in brain and gonads.The knockdown of foxl 2 by RNAi resulted in delay in the development of the female gonads.Conversely,no notable alterations were discerned in the gonads of C.carpio var.koi following the knockdown of cyp 19 b.The upregulation of sox 9 a,amh,and cyp 19 b following foxl 2 knockdown indicates that foxl 2 may play a pivotal role in gonadal development.Nevertheless,further investigation is required to ascertain the potential role of cyp 19 b.This study elucidated the role of foxl 2 and enhanced the understanding of the mechanisms of sex determination and gonadal development in C.carpio var.koi.
基金The Shanghai committee of Science and Technology(Grant No.10PJ1404800)the National Natural Science Foundation of China(Grant No.31072244)
文摘The means of survival of genomic dsRNA of reoviruses from dsRNA-triggered and Dicer-initiated RNAi pathway remains to be defined. The present study aimed to investigate the effect of Grass carp reovirus (GCRV) replication on the RNAi pathway of grass carp kidney cells (CIK). The dsRNA-triggered RNAi pathway was demonstrated unimpaired in CIK cells through RNAi assay. GCRV-specific siRNA was generated in CIK cells transfected with purified GCRV genomic dsRNA in Northern blot analysis; while in GCRV-infected CIK cells, no GCRV-specific siRNA could be detected. Infection and transfection experiments further indicated that replication of GCRV correlated with the increased transcription level of the Dicer gene and functional inhibition of in vitro synthesized egfp-siRNA in silencing the EGFP reporter gene. These data demonstrated that although only the genomic dsRNA of GCRV was sensitive to the cellular RNAi pathway, unidentified RNAi suppressor protein(s) might contribute to the survival of the viral genome and efficient viral replication.
基金This work was supported by the NationalPostdoctoral Science Foundation of China(No.2003034300)
文摘Objective: The effects on cell-cycle and p53 expression in hepatoma cell line SK-Hep-1 were explored by transfecting exogenous p53 small double stranded RNA (dsRNA) into the SK-Hep-1 cells. Methods: p53 dsRNA and EGFP dsRNA were synthesized. SK-Hep-1 (wtp53) cell line was transfected with 200 ng and 400 ng p53 dsRNA or EGFP and EGFP+EGFP dsRNA (as positive control) or 9% NaCl (as blank control) by liposome transfection technique. Flow cytometry was adopted to measure the effects of p53 dsRNA on cell cycle. Expression of p53 protein was detected by Western-Blotting at 48 h after transfecting p53 dsRNA. Results: The number of G0-G1 phase SK-Hep-1 cells, which were transfected with 200 ng p53 dsRNA, was decreased by 52.53% comparing with the control, and decreased by 50.29% (P<0.05) comparing with the positive control cells transfected with same dosage of EGFP+EGFP dsRNA. The number of S phase cells, which were transfected with 200 ng p53 dsRNA, was increased by 146.8% comparing with the control, and increased by 128.62% (P<0.05) comparing with the positive control cells transfected with same dosage of EGFP+EGFP dsRNA. The number of G2-M phase cells, which were transfected with 200 ng p53 dsRNA, was increased by 30.56% (P<0.05) comparing with the control, and increased by 21.63% (P>0.05) comparing with the positive control cells transfected with same dosage of EGFP+EGFP dsRNA. After 48 h, p53 protein expression was not detected in the SK-Hep-1 cells transfected with p53 dsRNA. Conclusion: p53 dsRNA can obviously improve the proliferation of SK-Hep-1 cells, and suppress p53 protein expression of SK-Hep-1 cells, the former may be related to of the latter.
基金Supported by a grant from the Natural Science Foundation of Shanghai （No. 03ZR14004）.
文摘Objective： Lung cancer has emerged as a leading cause of cancer death in the world. Current therapies are ineffective, thus new approaches are needed to improve the therapeutic ratio. RNA interference （RNAi） has shown promise in gene silencing in vitro, the potential of which in developing new methods for the therapy of non-small-cell lung cancer （NSCLC） needs to be further tested in vivo. In this study, chemically synthesized double-stranded RNA （dsRNA） targeting epidermal growth factor receptor （EGFR） was transfected into NSCLC cell line SPC-A1 cells and established the tumor burdened athymic nude mice model to investigate whether dsRNA could induce gene silencing in NSCLC cells in vivo. Methods： SPC-A1 was transfected with EGFR sequence-specific dsRNA formulated with Lipofectamine 2000. SPC-A1 cells （1 × 107/ mL） in 200 pL were injected s.c. into the left flank area of the mice to establish the tumor burdened athymic nude mice model. Calculate the tumor growth inhibition rate by measuring the diameter and the weight of the tumor. Immunohistochemistry and Westem blot were used to monitor the reduction in the production of the EGFR protein. Realtime RT-PCR was used to detect the silencing of the EGFR mRNA level. Results： It displayed that EGFR sequence specific dsRNA （dsRNA-EGFR） significantly inhibited the tumor growth in vivo. The tumor growth inhibition rate was 75.03%. The dsRNA-EGFR sequence specifically silenced EGFR with 53.6% of down-regulation of EGFR protein production and 32.3% of silencing of EGFR mRNA level. Conclusion： DsRNA-EGFR showed a blockbuster effect in downregulation of EGFR mRNA level and protein production, and inhibition of tumor growth in vivo.
基金supported by Key Research and Development Program of China National Tobacco Corporation (2021-150-110202102026)。
文摘Tobacco(Nicotiana tabacum)is a widely used platform for producing recombinant proteins for clinical applications.However,achieving mammalian-like glycosylation modifications in plantderived therapeutic proteins remains challenging,particularly in the case of fucosylation mediated by fucosyltransferases(FUTs).In this study,an RNA interference(RNAi)plasmid targeting the first exon ofα-1,3-fucosyltransferase 4(FUT4)gene was constructed,named as FUT4-RNAi.Using Agrobacterium-mediated transformation with EHA105 harboring the FUT4-RNAi plasmid,we obtained 29 regenerated tobacco lines,17 of which were confirmed as putatively positive by PCR.The mRNA transcript accumulation of the FUT4 gene was significantly reduced in 16 out of the 17 transgenic lines compared to the negative control,cv.Yunyan 87.Among these,11 lines(4^(#),6,7,11^(#),12^(#),15^(#),19^(#),22^(#),26^(#),28^(#) and 29^(#))showed FUT4 transcript levels below 25%of those in cv.Yunyan 87.Four lines(7^(#),12^(#),15^(#),and 29^(#))with the lowest mRNA levels were selected for further analysis by western blotting(WB)and enzyme-linked immunosorbent assay(ELISA).The results confirmed a significant decrease in FUT4 protein levels in these lines compared with that in cv.Yunyan 87,with line 29*showing less than 13%of the FUT4 protein content compared to cv.Yunyan 87.This study successfully developed a tobacco chassis with severely downregulated FUT4 expression,laying an important foundation for the production of human therapeutic proteins using a plant expression system.
基金(No. 2006AA10Z136) supported by National High-Tech R & D Program (863) of China
文摘Bone morphogenetic proteins(BMPs) play a critical role in the growth and steroidogenesis of granulosa cells(GCs).BMP signals act through membrane-bound heteromeric serine/threonine kinase receptors.Upon ligand binding,BMPs activate intracellular Smad proteins and regulate growth and apoptosis in various cell types.The objective of this study was to demonstrate the effects of BMP/Smad signal on growth and steroidogenesis of porcine GCs.A strategy of RNA interference(RNAi)-mediated 'gene silencing' of Smad4,a core molecule mediating the intracellular BMP/Smad signal transduction pathways,was used to interrupt endogenous BMP/Smad signaling.Results indicate that Smad4-small interfering RNA(siRNA) caused specific inhibition of Smad4 mRNA and protein expression after transfection.Interrupted endogenous BMP/Smad signaling significantly inhibited growth,and induced apoptosis of porcine GCs,while decreasing estradiol production.In addition,interrupted BMP/Smad signaling significantly(P<0.05) changed the expression of Cyclin D2,CDK4,Bcl-2,and Cyp19a1.These findings provide new insights into how BMP/Smad signaling regulates the growth and steroidogenesis of porcine GCs.
文摘Modern agribusiness plays a vital role in safeguarding and improving the production,quality,and quantity of food,feed,fiber,and fuel.Growing concerns over the impact of chemical pesticides on health and the environment have stimulated the industry to search for alternative and greener solutions.Over the last years,the RNA interference(RNAi)process has been identified as a very promising new approach to complement the arsenal of foliar spray,soil,or seed treatments applied as chemical and biological pest control agents,and of plant-incorporated protectants(PIPs).RNA-based active ingredients(AIs)possess a unique mode of action and can be implemented via both genetic modification(GM)and biocontrol approaches.RNA-based AIs promise to deliver the selectivity and sustainability desired in future crop protection agents.This is due to their utilization of a natural process to exert control and their high level of selectivity,which leads to reduced risk for non-target organisms(NTOs).This review discusses the advantages and limitations of RNA-based solutions in crop protection and recent research progress toward RNA-based biocontrols against the Colorado potato beetle(CPB),corn rootworm(CRW),and soy stink bug(SSB).Many challenges still exist on the road to the implementation of a broad range of RNA-based products and their widespread use and application.Despite these challenges,it can be expected that RNA-based AIs will become valuable new tools complementing the current arsenal of crop-protection solutions.
基金supported by Medical Science and Technology Research Project of Henan Province(142102310031)
文摘OBJECTIVE To explore the role of gecko crude peptides(GCPs)in the proliferation,apoptosis,migration and lymphangiogenesis of human hepatocellular carcinoma cells(Hep G2)and human lymphaticendothelial cells(HLECs)in vitro.METHODS The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay was used to evaluate the anti-proliferative effect of GCPs and si RNA-VEGF-C on Hep G2 cells,Hoechst 33258 staining and flow cytometry were performed to analyze cycle and apoptosis.The migration and invasion ability of cells were assayed by transwell chamber experiment and wound-healing assay.The protein and m RNA expressions of vascular endothelial growth factor-C(VEGF-C)and CXC chemokine receptor-4(CXCR4)were detected by q-PCR,immunofluorescence,Western blot.The protein expressions of the extracellular signal regulated kinase(ERKI/2),c-Jun N-terminal kinase(JNK),p38-mitogen activated protein kinases(p38 MAPK),serine/threonine kinase(Akt)and phosphatidylinositol-3-kinase(PI3K)were detected by western blot.The anti-lymphangiogenesis effect of GCPs on the HLECs was analyzed using an in vitro tube-formation assay.The protein and m RNA expressions of vascular endothelial growth factor receptor-3(VEGFR-3)and stromal cell-derived factor-1(SDF-1)were detected by q-PCR,Western blot.RESULTS GCPs and si RNA-VEGF-C inhibited Hep G2 proliferation,invasion and migration,and the most obvious inhibitory effect was both synergistic effects.Thus,GCPs suppressed HLECs proliferation,migration and tubelike structure formationin a dose-dependent manner,and had inhibitory effect of tumor-induced lymphangiogenesis in vitro.Additionally,we found that GCPs and si RNA-VEGF-C decreased the expressions of MMP-2,MMP-9,VEGF-C,CXCR4,phospho-ERK1/2,phospho-P38,phospho-JNK and PI3K in Hep G2 cells.Moreover,GCPs had a dose-dependent depressive effecton the expressions of VEGFR-3,SDF-1 in HLECs.CONCLUSION The low expression of VEGF-C mediated by si RNA-VEGF-C and GCPs inhibit tumor proliferation,invasion and migrationby suppressing the MAPK signaling pathway through reduced levels of VEGF-C,and GCPs inhibit tumor lymphangiogenesis by suppressing the CXCR4/SDF-1 signaling pathway through suppressed VEGF-C/VEGFR-3.
文摘Objective： To investigate the inhibitory effects of RNAi （ RNA interference, RNAi） expression vector on CXCR4 expression in prostate carcinoma cell lines. Methods： Small interference RNA （siRNA） expression vectors for CXCR4 gene were constructed and transfected into prostate carcinoma cell lines（PC-3m and LNCaP）with liposomes. T expression of CXCR4 was detected by RT-PCR and western blot. Results： T expression of CXCR4 mRNA and protein in the PC-3m and LNCaP cells was reduced by RNAi expression vectors. The inhibitory rate of CXCR4 mRNA expression in the PC-3m cells was 87.81% ± 10.20% ,56.10% ± 9.32% at the 24th hour and the 48th hour, compared with 56.93% ±8.78% ,49.24% ± 11.23% in LNCaP cells. The inhibitory rate of the expression of CXCR4 protein was 64.71% ± 6.68% ,58.66% ± 11.56% respectively. Conclusion： The expression of CXCR4 gene can effectively be inhibited by RNAi expression vectors.
文摘High mobility group A2（HMGA2） protein is a small nonhistone chromosomal protein that can modulate transcription of an ample number of genes.Many previous studies demonstrate that up-regulation of HMGA2 expression occurrs in many kinds of cancers including colorectal cancer,suggesting that HMGA2 might play a critical role in the progression of various tumors.However,the exact role of HMGA2 in colorectal cancer has not been determined.To verify the essential role of HMGA2 in the growth and invasiveness of colorectal cancer,HMGA2 expression was down-regulated by RNA interference（RNAi） in SW480 cells.We observed that the knockdown of HMGA2 led to the significant inhibition of proliferation and invasion of SW480 cells in vitro.These results suggest that HMGA2 might play a crucial role in the progression of colorectal cancer,and be a potential therapeutic target for human colorectal cancer.
文摘Objective：To investigate the silencing effects of recombinant adenovirus Ad-shRNA-MK on midkine（MK） gene in pancreatic cancer cells. Methods：Ad-shRNA-MK was used to infect pancreatic cancer BxPC-3 cells. Assays were conducted for knockdown of the MK gene on the day of infection and on the 1 ^st, 3^rd, 5^th, 7^th, and 9^th days post-infection by using immunocytochemistry, real-time RT-PCR, and Western blot analysis. Results：The adenoviral Ad-shRNA-PTN was constructed successfully, and infection was confirmed by electron microscopic observation. By using real-time RT-PCR, the inhibition rates of MK mRNA expression in the BxPC-3 cells were 20%, 80%, 55%, and 23% on the 1st, 3^th, 5^th, and 7^th days post-infection. Immunocytochemistry and Western blot analysis confirmed this effect at the gene product level. Conclusion：Efficient and specific knockdown of MK in pancreatic cancer cells by adenoviral Ad-shRNA-PTN is a potentially powerful tool for the study of gene therapy of pancreatic cancer nerve infiltration.
基金supported by the the National Natural Science Foundation of China(No.82003372)the Natural Science Foundation of Hubei Province(Nos.2018CFB747 and 2018CFB537)the Educational Commission of Hubei Province(Nos.B2017112 and B20181130),China.
文摘Objectives:In this study,we explored how adiponectin mediated urotensinⅡ(UⅡ)-induced tumor necrosis factor-α(TNF-α)andα-smooth muscle actin(α-SMA)expression and ensuing intracellular signaling pathways in adventitial fibroblasts(AFs).Methods:Growth-arrested AFs and rat tunica adventitia of vessels were incubated with UⅡand inhibitors of signal transduction pathways for 1-24 h.The cells were then harvested for TNF-αreceptor(TNF-α-R)messenger RNA(mRNA)and TNF-αprotein expression determination by reverse transcription-polymerase chain reaction(RT-PCR)and enzyme-linked immunosorbent assay(ELISA),respectively.Adiponectin and adiponectin receptor(adipoR)expression was measured by RT-PCR,quantitative real-time PCR(qPCR),immunohistochemical analysis,and cell counting kit-8(CCK-8)cell proliferation experiments.We then quantified TNF-αandα-SMA mRNA and protein expression levels by qPCR and immunofluorescence(IF)staining.RNA interference(RNAi)was used to explore the function of the adipoR genes.To investigate the signaling pathway,we applied western blotting(WB)to examine phosphorylation of adenosine 5’-monophosphate(AMP)-activated protein kinase(AMPK).In vivo,an adiponectin(APN)-knockout(APN-KO)mouse model mimicking adventitial inflammation was generated to measure TNF-αandα-SMA expression by application of qPCR and IF,with the goal of gaining a comprehensive atlas of adiponectin in vascular remodeling.Results:In both cells and tissues,UⅡpromoted TNF-αprotein and TNF-α-R secretion in a dose-and time-dependent manner via Rho/protein kinase C(PKC)pathway.We detected marked expression of adipoR1,T-cadherin,and calreticulin as well as a moderate presence of adipoR2 in AFs,while no adiponectin was observed.Globular adiponectin(gAd)fostered the growth of AFs,and acted in concert with UⅡto induceα-SMA and TNF-αthrough the adipoR1/T-cadherin/calreticulin/AMPK pathway.In AFs,gAd and UⅡsynergistically induced AMPK phosphorylation.In the adventitial inflammation model,APN deficiency up-regulated the expression ofα-SMA,UⅡreceptor(UT),and UⅡwhile inhibiting TNF-αexpression.Conclusions:From the results of our study,we can speculate that UⅡinduces TNF-αprotein and TNF-α-R secretion in AFs and rat tunica adventitia of vessels via the Rho and PKC signal transduction pathways.Thus,it is plausible that adiponectin is a major player in adventitial progression and could serve as a novel therapeutic target for cardiovascular disease administration.
基金Supported grants from National Natural Science Foundation of China (No.30772547)Doctoral Fund of Ministry of Education of China (No.20060631013)
文摘Objective:Survivin has attracted abundant interest in tumor research since it was discovered in 1997.But several studies indicated that the relationship between survivin expression and tumor behavior is still not fully understood.So our main aim was to observe the localization of survivin in colon cancer and its influence on the colon cancer biocharacteristics.Methods:We screened the expression and localization of survivin in SW480 by immunofluorescence and immunocytochem-istry.Then, we constructed the recombinant adenovirus (Ad-survivin/shRNA), which contained the short hairpin RNA (shRNA) of survivin and transfected it into SW480 cells.We detected survivin gene expression after shRNA interference by RT-PCR and Western blot, and its influence on the proliferation, apoptosis and cell cycle was analyzed by MTT, colony-formation, and flow cytometry.Results:Survivin was expressed at a high level in SW480 cells and the sub-cellular localization was in the cytoplasm.Recombinant adenovirus could suppress survivin expression efficiency and inhibit proliferation, induce apoptosis, and affected the mitosis in vitro.Conclusion:Survivin plays an important role in controlling tumor growth by a variety of molecular regulatory mechanisms.Cytoplasmic survivin silence could induce apoptosis, effect the mitotic cycle and inhibit cell proliferation.
基金supported by the Program of National Science Fund for Distinguished Young Scholars of China(No.81925032)Key Program of National Natural Science Foundation of China(No.82130098)National Natural Science Foundation of China(No.82304232)。
文摘Objective: Genome-wide association studies(GWAS) have identified over 150 risk loci linked to colorectal cancer(CRC), including the 17p13.3 locus with the tag single nucleotide polymorphism(SNP) rs12603526 in the Asian population. However, the specific causal gene and the functional regulatory mechanisms in this region remain unresolved, necessitating further investigation to elucidate the underlying mechanisms of CRC.Methods: We employed an RNA interference-based functional approach to identify genes critical for CRC cell proliferation at the GWAS locus 17p13.3. Bioinformatic fine-mapping analysis was conducted to prioritize causal variants. A large-scale study involving 7,013 cases and 7,329 controls from a Chinese population, along with another cohort of 5,158 cases and 20,632 controls from the UK Biobank, was performed to validate the association between the candidate variant and the gene. A series of biological experiments was conducted to explore the function of the candidate gene and its regulatory mechanisms.Results: We identified FAM57A as a key oncogene that promotes CRC cell proliferation, and confirmed its carcinogenic role through in vitro proliferation assays. The variant rs526835 was prioritized as a causal candidate for CRC risk, located in a functional region with enhancer properties, and showed a significant quantitative association with FAM57A expression. The rs526835 [T] variant was associated with a 1.17-fold increase in CRC risk [95%confidence interval(95% CI): 1.11-1.23, P=1.23×10^(−9)] in the large-scale Chinese cohort, which was further corroborated in the UK Biobank cohort. Mechanistically, we demonstrated that rs526835 enhances a promoterenhancer interaction mediated by the transcription factor JUN, leading to increased expression of FAM57A.Conclusions: We reveal the underlying mechanisms of CRC predisposition at the GWAS locus 17p13.3.Additionally, our findings highlight the critical role of FAM57A in CRC pathogenesis and introduce a novel enhancer-promoter interaction between FAM57A and rs526835, which could inform future precision prevention and personalized cancer therapies.
文摘Objective:The aim of this study was to study the changes of SGC-7901 cells transfecting small hairpin RNA(shRNA) targeted Bcl-2 and celecoxib in vitro.Methods:To use the effective siRNA targeted to Bcl-2 by Lipofectamine 2000,the rate of cell growth inhibition of all groups was detected by multiply-table tournament(MTT) method,apoptosis rate was examined by flow cytometry and the expression of Bcl-2 was assayed by Elisa.Results:After siRNA was transfected to SGC-7901 cells,the rate of cell growth inhibition was increased,the joint group was higher by flow cytometry and the expression of Bcl-2 was lower by Elisa.Conclusion:The growth of SGC-7901 cells which was transfected siRNA has been inhibited,and the sensitivity to celecoxib has been increased.
文摘The paper aimed to study the effect of lysyl oxidase-like 2 （LOXL2） on hepatocellular carcinoma （HCC） and explore the biological mechanisms of tumorigenicity and progression in HCC. The authors used four HCC cell lines to identify LOXL2. A lentiviral vector containing LOXL2-siRNA was constructed to silence the LOXL2 gene in SMMC-7721 cell line, and mRNA of the target gene was detected by real-time polymerase chain reaction （RT-PCR）. The effect of LOXL2 silencing on the growth of SMMC-7721 cells was explored with flow cytometry profiling and BrdU labeling. Downstream genes of LOXL2 were selected by microarray and verified by Western Blotting. In the results, LOXL2 expression was significantly up-regulated in four types of HCC cell lines, therefore, SMMC-7721 cell line was selected for further exploration. When SMMC-7721 cell line was infected with LOXL2-siRNA, the expression of LOXL2 mRNA decreased. The silencing of LOXL2 resulted in the cell cycle arrest at the G 1-phase, the increased apoptosis and the decreased growth of SMMC-7721 cells on the indicated days by BrdU. Moreover, the MDM2, BIRC3, CDC42, FOS and TGFBR2 genes were selected and verified to be the downstream genes of LOXL2. In conclusion, LOXL2 contributes to the genesis and progression of HCC cells and works by regulating downstream genes of LOXL2 in certain pathways. Therefore, LOXL2 may play an important role in the progression and prognosis of HCC.
