The first internal transcribed spacer(ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced wi...The first internal transcribed spacer(ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced with automated fluorescent sequencing. The sequences of ITS1 ranged from 193 bp to 218 bp in size and G/C content varied from 69.3% to 72.7%. In pairwise comparisons among the five taxa, sequence site divergence ranged from 1.5% to 10.6%. Phylogenetic analysis of ITS1 sequences using Wagner parsimony generated a single well resolved tree, which revealed that Oryza rufipogon was much more closely related to cultivated rice species than to the other two wild species. Oryza granulata was less closely related to either cultivated rice species or the other two wild species, and might be a unique and isolated taxon in the genus Oryza. The phylogenetic relationships of the three wild rice species and two cultivated rice subspecies inferred from ITS1 sequences is highly concordant with those based on the molecular evidence from isozyme, chloroplast DNA (cpDNA), mitochondrial DNA(mtDNA) and nuclear DNA (nDNA) of the genus Oryza .展开更多
[Objective] The aim was to establish a molecular biological method for identifying coccidium species.[Method]First,the pure species of Eimeria intestinalis was isolated by using single-oocyst isolation technique.Then,...[Objective] The aim was to establish a molecular biological method for identifying coccidium species.[Method]First,the pure species of Eimeria intestinalis was isolated by using single-oocyst isolation technique.Then,according to the 18s rDNA and 5.8s rDNA sequences of Eimeria coccidia published in GenBank,a pair of specific primers was designed and synthesized to amplify the internal transcribed spacer 1（ITS-1）.Finally,the PCR products were sent for sequencing.[Result]The pure species of E.intestinalis was isolated and the result of agarose gel electrophoresis showed that the PCR product was 434 bp,and at least 27-sporulated oocysts could be detected.[Conclusion]The research will provide a basis for accurate identification of coccidium species and strains.展开更多
For the solid-solid transformation from formⅡto formⅠof isotactic polybutene-1(iPB),the temperature dependence of formⅠnucleation and growth was deemed to control the transformation process.However,the relationship...For the solid-solid transformation from formⅡto formⅠof isotactic polybutene-1(iPB),the temperature dependence of formⅠnucleation and growth was deemed to control the transformation process.However,the relationship between formⅠformation and formⅡdisappearance in the transformation process is not clear.In this work,the spontaneous crystal transformation from formⅡtoⅠof iPB with 81 mol%mmmm sequence concentration is studied firstly by tracking the two processes,the decay of formⅡand the yielding of formⅠin a wide range of temperature spanning from 0℃to 50℃and in a long transformation time ranging from 5 min to 65 days with in situ FTIR and WAXD.Unlike the literature reports,the decay rate of formⅡis firstly found to be lower than the yielding rate of formⅠat all studied temperatures,especially at low transition temperature.This is attributed to the amorphous chains which locate near crystal lamella participating into the nucleation of formⅡ.The regular chain folding and growth of i PB formⅠfrom amorphous chains containing short isotactic sequences also lead to an increase in crystallinity of formⅠcompared with that of initial formⅡcrystallized at 60℃.An increase in the annealing temperature results in decrease in crystallinity and increase in lamellae thickness of i PB formⅠ.展开更多
For a long time,classification of Demodex mites has been based mainly on their hosts and phenotypic characteristics.A new subspecies of Demodex folliculorum has been proposed,but not confirmed.Here,cox1 partial sequen...For a long time,classification of Demodex mites has been based mainly on their hosts and phenotypic characteristics.A new subspecies of Demodex folliculorum has been proposed,but not confirmed.Here,cox1 partial sequences of nine isolates of three Demodex species from two geographical sources(China and Spain) were studied to conduct molecular identification of D.folliculorum.Sequencing showed that the mitochondrial cox1 fragments of five D.folliculorum isolates from the facial skin of Chinese individuals were 429 bp long and that their sequence identity was 97.4%.The average sequence divergence was 1.24% among the five Chinese isolates,0.94% between the two geographical isolate groups(China(5) and Spain(1)),and 2.15% between the two facial tissue sources(facial skin(6) and eyelids(1)).The genetic distance and rate of third-position nucleotide transition/transversion were 0.0125,2.7(3/1) among the five Chinese isolates,0.0094,3.1(3/1) between the two geographical isolate groups,and 0.0217,4.4(3/1) between the two facial tissue sources.Phylogenetic trees showed that D.folliculorum from the two geographical isolate groups did not form sister clades,while those from different facial tissue sources did.According to the molecular characteristics,it appears that subspecies differentiation might not have occurred and that D.folliculorum isolates from the two geographical sources are of the same population.However,population differentiation might be occurring between isolates from facial skin and eyelids.展开更多
Herein, we report a very high content of simple sequence repeats (SSRs) covering 66.12% of the herpes simplex virus type 1 (HSV-1) genome when a low threshold is adopted to define SSRs, indicating that repeat sequence...Herein, we report a very high content of simple sequence repeats (SSRs) covering 66.12% of the herpes simplex virus type 1 (HSV-1) genome when a low threshold is adopted to define SSRs, indicating that repeat sequence is a very important character of the HSV-1 genome. The repeats with two iterations account for 68.33% of the total repeats. In reality, the genome of HSV-1 is prone to form shorter repeat sequences. For mono-, di- and trinucleotide repeats, the repeat numbers decreased with the increase of repeats iterations, implicating that the formation tendency of SSRs might be from low iterations to high iterations. The high iterations SSRs might have subjected to strong selected pressure and survived to perform different functions. The analysis suggested that the repeats formation may be an essential evolutionary driving force for the HSV-1 genome, and the results might be helpful for studying the genome structure, repeats genesis and genome evolution of HSV-1.展开更多
N6-methyladenosine(m^(6)A)modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function.The m^(6)A reader YTHDF1 has been shown to enhance the transl...N6-methyladenosine(m^(6)A)modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function.The m^(6)A reader YTHDF1 has been shown to enhance the translational efficiency of m^(6)A-modified mRNAs in the brain and is essential for learning and memory.However,its role in the mature retina remains unclear.Herein,we report a novel role of Ythdf1 in the maintenance of retinal function using a genetic knockout model.Loss of Ythdf1 resulted in impaired scotopic electroretinogram(ERG)responses and progressive retinal degeneration.Detailed analyses of rod photoreceptors confirmed substantial degenerative changes in the absence of ciliary defects.Single-cell RNA sequencing revealed comprehensive molecular alterations across all retinal cell types in Ythdf1-deficient retinas.Integrative analysis of methylated RNA immunoprecipitation(MeRIP)sequencing and RIP sequencing identified Tulp1 and Dhx38,two inheritable retinal degeneration disease-associated gene homologs,as direct targets of YTHDF1 in the retina.Specifically,YTHDF1 recognized and bound m^(6)A-modified Tulp1 and Dhx38 mRNA at the coding sequence(CDS),enhancing their translational efficiency without altering mRNA levels.Collectively,these findings highlight the essential role of YTHDF1 in preserving visual function and reveal a novel regulatory mechanism of m^(6)A reader proteins in retinal degeneration,identifying potential therapeutic targets for severe retinopathies.展开更多
Head and neck cutaneous squamous cell carcinoma(HNCSCC)remains underexplored compared to oropharyngeal squamous cell carcinoma,particularly in relation to human papillomavirus(HPV)and molecular markers such as p16 and...Head and neck cutaneous squamous cell carcinoma(HNCSCC)remains underexplored compared to oropharyngeal squamous cell carcinoma,particularly in relation to human papillomavirus(HPV)and molecular markers such as p16 and p53.While p16 is a well-established surrogate for HPV in oropharyngeal cancer,our review highlights its unreliable role in HNCSCC,where positivity is instead associated with recurrence and metastasis.Similarly,p53 illustrates a dual role-wild-type as a genomic safeguard,mutated as an oncogenic driver-complicating prognostication.Methodological considerations,including the limitations of immunohistochemistry for HPV detection,underscore the need for multi-method and molecular validation in future studies.Ultraviolet radiation is posited as a key modifier of p16 function,decoupling expression from tumor suppression.To contextualize these findings,we draw parallels to glioblastoma(GBM),where subclonal evolution,p53 dysfunction,and intratumoral heterogeneity drive relapse despite aggressive multimodal therapies.GBM exemplifies how bulk-level biomarker generalizations often obscure dynamic cellular ecosystems,reinforcing the necessity of single-cell and spatial approaches.Multi-omics integration-encompassing genome,transcriptome,proteome,and tumor microenvironment mapping-coupled with single-cell RNA sequencing and spatial transcriptomics,offers a path forward for resolving subclonal dynamics in both HNCSCC and GBM.These technologies provide the resolution needed to track tumor-immunestromal co-evolution,identify therapy-resistant clones,and anticipate recurrence.We argue for a N-of-1,patient-and cell-centric paradigm that reframes biomarkers not as static surrogates but as dynamic readouts of cancer evolution across time and tissue contexts.Conceptually,we propose kinetic and microenvironmental frameworks(e.g.,“load-and-lock”barriers;dormancy and immunesynapse stabilization)as hypothesis-generating avenues to stall clonal handoffs and improve outcome prediction.Together,these perspectives argue for revised biomarker frameworks in HNCSCC and ethically inclusive,mechanism-anchored studies that bridge discovery with individualized care.By bridging insights from HNCSCC with the lessons of GBM,this review underscores the need for ethically inclusive,mechanistically informed frameworks that integrate subclonal evolution,biomarker re-interpretation,and precision-personalized hybrid models.Such an approach will be essential for advancing from one-size-fits-all strategies to individualized lifetime cancer care.展开更多
目的探讨肺腺癌细胞恶性T细胞扩增序列1(MCTS1)介导骨髓源性抑制细胞(MDSCs)趋化的作用机制。方法利用生物信息学(http://timer.cistrome.org/#tab-8258-2)分析癌症基因组图谱(TCGA)中肺腺癌组织(n=515)MCTS1的表达水平与MDSCs浸润之间...目的探讨肺腺癌细胞恶性T细胞扩增序列1(MCTS1)介导骨髓源性抑制细胞(MDSCs)趋化的作用机制。方法利用生物信息学(http://timer.cistrome.org/#tab-8258-2)分析癌症基因组图谱(TCGA)中肺腺癌组织(n=515)MCTS1的表达水平与MDSCs浸润之间的相关性;利用IL-6+粒细胞-巨噬细胞集落刺激因子(GM-CSF)共同诱导人外周血单个核细胞(PBMCs)中MDSCs的生成,流式细胞术检测MDSCs生成比例,CD33磁珠分选MDSCs;收集肺腺癌SPC-A1细胞野生型以及MCTS1敲减和过表达SPC-A1细胞的无血清细胞上清液,利用Transwell小室进行MDSCs的趋化试验并计算趋化指数;羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记CD8^(+)T细胞与发生趋化的MDSCs细胞共培养,流式细胞术检测CD8^(+)T细胞的增殖水平;实时荧光定量PCR(qRT-PCR)测定MDSCs相关趋化因子的mRNA水平;流式多因子芯片技术检测细胞上清液中C-X-C基序趋化因子配体1(CXCL1)的含量;Western blot检测janus激酶2(JAK2)的表达水平以及信号转导子和转录激活子1(STAT1)的活化水平。结果肺腺癌组织MCTS1与MDSCs浸润程度呈正相关(Spearmanρ=0.226,P<0.001);与空白对照组比较,IL-6和GM-CSF共同诱导PBMCs后可显著增加MDSCs的比例(4.95±1.03 vs 0.97±0.24,t=6.54,P<0.01);MCTS1过表达的SPC-A1细胞上清液对MDSCs的趋化指数较对照组显著升高(4.88±0.99 vs 2.94±0.27,q=6.29,P<0.01),而经MCTS1敲减的细胞上清液可使MDSCs的趋化指数显著降低(1.50±0.17 vs 2.94±0.27,q=4.69,P<0.05);MCTS1过表达的SPC-A1细胞中CXCL1 mRNA(2.79±0.16 vs 1.00±0.08,q=28.94,P<0.001)和上清液中蛋白质水平(78.20±6.16 vs 37.50±3.31,q=16.83,P<0.001)均显著升高,而敲减MCTS1则可抑制CXCL1 mRNA(0.53±0.03 vs 1.00±0.08,q=7.64,P<0.01)和蛋白质水平(17.33±1.96 vs 37.50±3.31,q=8.34,P<0.01);此外,与对照组比较,MCTS1过表达组JAK2蛋白的表达水平(2.11±0.15 vs 1.00±0.05,q=19.48,P<0.001)和STAT1的活化水平(2.10±0.19 vs 1.00±0.10,q=15.02,P<0.001)显著升高,而MCTS1敲减组JAK2蛋白水平(0.56±0.06 vs 1.00±0.05,q=7.61,P<0.01)和STAT1的活化水平均显著降低(0.46±0.07 vs 1.00±0.05,q=7.45,P<0.01)。结论肺腺癌细胞MCTS1通过JAK2-STAT1信号通路调控CXCL1的表达,并介导MDSCs的趋化。展开更多
Objectives:Philadelphia chromosome-positive B-cell acute lymphoblastic leukemia and Philadelphia-like B-cell acute lymphoblastic leukemia(Ph+/Ph-like ALL)constitute the majority of relapsed/refractory B-ALL(R/R B-ALL)...Objectives:Philadelphia chromosome-positive B-cell acute lymphoblastic leukemia and Philadelphia-like B-cell acute lymphoblastic leukemia(Ph+/Ph-like ALL)constitute the majority of relapsed/refractory B-ALL(R/R B-ALL)cases,highlighting an urgent need to discover new therapeutic targets.This study aims to elucidate the mechanisms underlying poor prognosis in Ph+/Ph-like ALL through transcriptome sequencing and functional cytological assays,with the goal of informing new clinical treatment strategies.Results:Transcriptomic analysis of Ph+/Ph-like ALL patients revealed that low expression of P2X Purinoceptor 1(P2RX1)was associated with unfavorable outcomes.Specifically,patients with poor prognosis and low P2RX1 expression exhibited downregulation of genes involved in energy and calcium metabolism pathways,along with upregulation of genes governing key cellular processes such as cell proliferation(e.g.,MYC),cell cycle progression(e.g.,CCND2),and apoptosis inhibition(e.g.,DASP6).Cellular experiments demonstrated that SUP-B15 cells overexpressing P2RX1 displayed elevated intracellular levels of ATP,calcium,and glucose,together with enhanced glycolytic capacity,compared to empty vector controls.Treatment of SUP-B15 cells with dexamethasone(Dex),Imatinib,or their combination significantly suppressed proliferation and promoted apoptosis,which was accompanied by increases in intracellular ATP,calcium,and glucose.Moreover,exogenous ATP administration(a P2RX1 agonist)enhanced apoptosis and inhibited proliferation in control cells.Conversely,treatment with NF449(a P2RX1 inhibitor)increased proliferation in both P2RX1-overexpressing and control SUP-B15 cells.Conclusion:Our findings indicate that P2RX1 may exert this function through modulating energy metabolism and calcium homeostasis,resulting in elevated intracellular calcium levels.Sustained elevation of calcium promotes apoptosis,whereas exogenous ATP activates P2RX1,enhances calcium influx,and attenuates the suppression of apoptosis associated with P2RX1 underexpression,ultimately correlating with improved treatment response.展开更多
基金The project supported by the Grant from Presidentof Chinese Academy of Sciences
文摘The first internal transcribed spacer(ITS1) of nuclear ribosomal DNA of three wild rice species and two subspecies of cultivated rice, which are distributed in China, was amplified using PCR technique and sequenced with automated fluorescent sequencing. The sequences of ITS1 ranged from 193 bp to 218 bp in size and G/C content varied from 69.3% to 72.7%. In pairwise comparisons among the five taxa, sequence site divergence ranged from 1.5% to 10.6%. Phylogenetic analysis of ITS1 sequences using Wagner parsimony generated a single well resolved tree, which revealed that Oryza rufipogon was much more closely related to cultivated rice species than to the other two wild species. Oryza granulata was less closely related to either cultivated rice species or the other two wild species, and might be a unique and isolated taxon in the genus Oryza. The phylogenetic relationships of the three wild rice species and two cultivated rice subspecies inferred from ITS1 sequences is highly concordant with those based on the molecular evidence from isozyme, chloroplast DNA (cpDNA), mitochondrial DNA(mtDNA) and nuclear DNA (nDNA) of the genus Oryza .
基金Supported by the Program of National Rabbit Industrial Production Technology System of Ministry of Agriculture~~
文摘[Objective] The aim was to establish a molecular biological method for identifying coccidium species.[Method]First,the pure species of Eimeria intestinalis was isolated by using single-oocyst isolation technique.Then,according to the 18s rDNA and 5.8s rDNA sequences of Eimeria coccidia published in GenBank,a pair of specific primers was designed and synthesized to amplify the internal transcribed spacer 1（ITS-1）.Finally,the PCR products were sent for sequencing.[Result]The pure species of E.intestinalis was isolated and the result of agarose gel electrophoresis showed that the PCR product was 434 bp,and at least 27-sporulated oocysts could be detected.[Conclusion]The research will provide a basis for accurate identification of coccidium species and strains.
基金financially supported by the Major Science and Technology Innovation Project of Shandong Province(No.2019JZZY010352)Natural Science Foundation of Shandong Province(ZR2019MB072)Taishan Scholar Program
文摘For the solid-solid transformation from formⅡto formⅠof isotactic polybutene-1(iPB),the temperature dependence of formⅠnucleation and growth was deemed to control the transformation process.However,the relationship between formⅠformation and formⅡdisappearance in the transformation process is not clear.In this work,the spontaneous crystal transformation from formⅡtoⅠof iPB with 81 mol%mmmm sequence concentration is studied firstly by tracking the two processes,the decay of formⅡand the yielding of formⅠin a wide range of temperature spanning from 0℃to 50℃and in a long transformation time ranging from 5 min to 65 days with in situ FTIR and WAXD.Unlike the literature reports,the decay rate of formⅡis firstly found to be lower than the yielding rate of formⅠat all studied temperatures,especially at low transition temperature.This is attributed to the amorphous chains which locate near crystal lamella participating into the nucleation of formⅡ.The regular chain folding and growth of i PB formⅠfrom amorphous chains containing short isotactic sequences also lead to an increase in crystallinity of formⅠcompared with that of initial formⅡcrystallized at 60℃.An increase in the annealing temperature results in decrease in crystallinity and increase in lamellae thickness of i PB formⅠ.
基金Project(No. 81271856)supported by the National Natural Science Foundation of China
文摘For a long time,classification of Demodex mites has been based mainly on their hosts and phenotypic characteristics.A new subspecies of Demodex folliculorum has been proposed,but not confirmed.Here,cox1 partial sequences of nine isolates of three Demodex species from two geographical sources(China and Spain) were studied to conduct molecular identification of D.folliculorum.Sequencing showed that the mitochondrial cox1 fragments of five D.folliculorum isolates from the facial skin of Chinese individuals were 429 bp long and that their sequence identity was 97.4%.The average sequence divergence was 1.24% among the five Chinese isolates,0.94% between the two geographical isolate groups(China(5) and Spain(1)),and 2.15% between the two facial tissue sources(facial skin(6) and eyelids(1)).The genetic distance and rate of third-position nucleotide transition/transversion were 0.0125,2.7(3/1) among the five Chinese isolates,0.0094,3.1(3/1) between the two geographical isolate groups,and 0.0217,4.4(3/1) between the two facial tissue sources.Phylogenetic trees showed that D.folliculorum from the two geographical isolate groups did not form sister clades,while those from different facial tissue sources did.According to the molecular characteristics,it appears that subspecies differentiation might not have occurred and that D.folliculorum isolates from the two geographical sources are of the same population.However,population differentiation might be occurring between isolates from facial skin and eyelids.
文摘Herein, we report a very high content of simple sequence repeats (SSRs) covering 66.12% of the herpes simplex virus type 1 (HSV-1) genome when a low threshold is adopted to define SSRs, indicating that repeat sequence is a very important character of the HSV-1 genome. The repeats with two iterations account for 68.33% of the total repeats. In reality, the genome of HSV-1 is prone to form shorter repeat sequences. For mono-, di- and trinucleotide repeats, the repeat numbers decreased with the increase of repeats iterations, implicating that the formation tendency of SSRs might be from low iterations to high iterations. The high iterations SSRs might have subjected to strong selected pressure and survived to perform different functions. The analysis suggested that the repeats formation may be an essential evolutionary driving force for the HSV-1 genome, and the results might be helpful for studying the genome structure, repeats genesis and genome evolution of HSV-1.
基金supported by the National Natural Science Foundation of China(82371083,82471100,82121003,82271084)Program of Science and Technology International Cooperation Project of Qinghai province(China)(2022-HZ-814)。
文摘N6-methyladenosine(m^(6)A)modification of mRNA is a critical post-transcriptional regulatory mechanism that modulates mRNA metabolism and neuronal function.The m^(6)A reader YTHDF1 has been shown to enhance the translational efficiency of m^(6)A-modified mRNAs in the brain and is essential for learning and memory.However,its role in the mature retina remains unclear.Herein,we report a novel role of Ythdf1 in the maintenance of retinal function using a genetic knockout model.Loss of Ythdf1 resulted in impaired scotopic electroretinogram(ERG)responses and progressive retinal degeneration.Detailed analyses of rod photoreceptors confirmed substantial degenerative changes in the absence of ciliary defects.Single-cell RNA sequencing revealed comprehensive molecular alterations across all retinal cell types in Ythdf1-deficient retinas.Integrative analysis of methylated RNA immunoprecipitation(MeRIP)sequencing and RIP sequencing identified Tulp1 and Dhx38,two inheritable retinal degeneration disease-associated gene homologs,as direct targets of YTHDF1 in the retina.Specifically,YTHDF1 recognized and bound m^(6)A-modified Tulp1 and Dhx38 mRNA at the coding sequence(CDS),enhancing their translational efficiency without altering mRNA levels.Collectively,these findings highlight the essential role of YTHDF1 in preserving visual function and reveal a novel regulatory mechanism of m^(6)A reader proteins in retinal degeneration,identifying potential therapeutic targets for severe retinopathies.
文摘Head and neck cutaneous squamous cell carcinoma(HNCSCC)remains underexplored compared to oropharyngeal squamous cell carcinoma,particularly in relation to human papillomavirus(HPV)and molecular markers such as p16 and p53.While p16 is a well-established surrogate for HPV in oropharyngeal cancer,our review highlights its unreliable role in HNCSCC,where positivity is instead associated with recurrence and metastasis.Similarly,p53 illustrates a dual role-wild-type as a genomic safeguard,mutated as an oncogenic driver-complicating prognostication.Methodological considerations,including the limitations of immunohistochemistry for HPV detection,underscore the need for multi-method and molecular validation in future studies.Ultraviolet radiation is posited as a key modifier of p16 function,decoupling expression from tumor suppression.To contextualize these findings,we draw parallels to glioblastoma(GBM),where subclonal evolution,p53 dysfunction,and intratumoral heterogeneity drive relapse despite aggressive multimodal therapies.GBM exemplifies how bulk-level biomarker generalizations often obscure dynamic cellular ecosystems,reinforcing the necessity of single-cell and spatial approaches.Multi-omics integration-encompassing genome,transcriptome,proteome,and tumor microenvironment mapping-coupled with single-cell RNA sequencing and spatial transcriptomics,offers a path forward for resolving subclonal dynamics in both HNCSCC and GBM.These technologies provide the resolution needed to track tumor-immunestromal co-evolution,identify therapy-resistant clones,and anticipate recurrence.We argue for a N-of-1,patient-and cell-centric paradigm that reframes biomarkers not as static surrogates but as dynamic readouts of cancer evolution across time and tissue contexts.Conceptually,we propose kinetic and microenvironmental frameworks(e.g.,“load-and-lock”barriers;dormancy and immunesynapse stabilization)as hypothesis-generating avenues to stall clonal handoffs and improve outcome prediction.Together,these perspectives argue for revised biomarker frameworks in HNCSCC and ethically inclusive,mechanism-anchored studies that bridge discovery with individualized care.By bridging insights from HNCSCC with the lessons of GBM,this review underscores the need for ethically inclusive,mechanistically informed frameworks that integrate subclonal evolution,biomarker re-interpretation,and precision-personalized hybrid models.Such an approach will be essential for advancing from one-size-fits-all strategies to individualized lifetime cancer care.
文摘目的探讨肺腺癌细胞恶性T细胞扩增序列1(MCTS1)介导骨髓源性抑制细胞(MDSCs)趋化的作用机制。方法利用生物信息学(http://timer.cistrome.org/#tab-8258-2)分析癌症基因组图谱(TCGA)中肺腺癌组织(n=515)MCTS1的表达水平与MDSCs浸润之间的相关性;利用IL-6+粒细胞-巨噬细胞集落刺激因子(GM-CSF)共同诱导人外周血单个核细胞(PBMCs)中MDSCs的生成,流式细胞术检测MDSCs生成比例,CD33磁珠分选MDSCs;收集肺腺癌SPC-A1细胞野生型以及MCTS1敲减和过表达SPC-A1细胞的无血清细胞上清液,利用Transwell小室进行MDSCs的趋化试验并计算趋化指数;羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)标记CD8^(+)T细胞与发生趋化的MDSCs细胞共培养,流式细胞术检测CD8^(+)T细胞的增殖水平;实时荧光定量PCR(qRT-PCR)测定MDSCs相关趋化因子的mRNA水平;流式多因子芯片技术检测细胞上清液中C-X-C基序趋化因子配体1(CXCL1)的含量;Western blot检测janus激酶2(JAK2)的表达水平以及信号转导子和转录激活子1(STAT1)的活化水平。结果肺腺癌组织MCTS1与MDSCs浸润程度呈正相关(Spearmanρ=0.226,P<0.001);与空白对照组比较,IL-6和GM-CSF共同诱导PBMCs后可显著增加MDSCs的比例(4.95±1.03 vs 0.97±0.24,t=6.54,P<0.01);MCTS1过表达的SPC-A1细胞上清液对MDSCs的趋化指数较对照组显著升高(4.88±0.99 vs 2.94±0.27,q=6.29,P<0.01),而经MCTS1敲减的细胞上清液可使MDSCs的趋化指数显著降低(1.50±0.17 vs 2.94±0.27,q=4.69,P<0.05);MCTS1过表达的SPC-A1细胞中CXCL1 mRNA(2.79±0.16 vs 1.00±0.08,q=28.94,P<0.001)和上清液中蛋白质水平(78.20±6.16 vs 37.50±3.31,q=16.83,P<0.001)均显著升高,而敲减MCTS1则可抑制CXCL1 mRNA(0.53±0.03 vs 1.00±0.08,q=7.64,P<0.01)和蛋白质水平(17.33±1.96 vs 37.50±3.31,q=8.34,P<0.01);此外,与对照组比较,MCTS1过表达组JAK2蛋白的表达水平(2.11±0.15 vs 1.00±0.05,q=19.48,P<0.001)和STAT1的活化水平(2.10±0.19 vs 1.00±0.10,q=15.02,P<0.001)显著升高,而MCTS1敲减组JAK2蛋白水平(0.56±0.06 vs 1.00±0.05,q=7.61,P<0.01)和STAT1的活化水平均显著降低(0.46±0.07 vs 1.00±0.05,q=7.45,P<0.01)。结论肺腺癌细胞MCTS1通过JAK2-STAT1信号通路调控CXCL1的表达,并介导MDSCs的趋化。
基金supported by Guangdong Province Basic and Applied Basic Research Fund Project(2023A1515220104)Open Fund of Key Laboratory of Hepatoaplenic Surgery,Ministry of Education(Award Number:GPKF202407).
文摘Objectives:Philadelphia chromosome-positive B-cell acute lymphoblastic leukemia and Philadelphia-like B-cell acute lymphoblastic leukemia(Ph+/Ph-like ALL)constitute the majority of relapsed/refractory B-ALL(R/R B-ALL)cases,highlighting an urgent need to discover new therapeutic targets.This study aims to elucidate the mechanisms underlying poor prognosis in Ph+/Ph-like ALL through transcriptome sequencing and functional cytological assays,with the goal of informing new clinical treatment strategies.Results:Transcriptomic analysis of Ph+/Ph-like ALL patients revealed that low expression of P2X Purinoceptor 1(P2RX1)was associated with unfavorable outcomes.Specifically,patients with poor prognosis and low P2RX1 expression exhibited downregulation of genes involved in energy and calcium metabolism pathways,along with upregulation of genes governing key cellular processes such as cell proliferation(e.g.,MYC),cell cycle progression(e.g.,CCND2),and apoptosis inhibition(e.g.,DASP6).Cellular experiments demonstrated that SUP-B15 cells overexpressing P2RX1 displayed elevated intracellular levels of ATP,calcium,and glucose,together with enhanced glycolytic capacity,compared to empty vector controls.Treatment of SUP-B15 cells with dexamethasone(Dex),Imatinib,or their combination significantly suppressed proliferation and promoted apoptosis,which was accompanied by increases in intracellular ATP,calcium,and glucose.Moreover,exogenous ATP administration(a P2RX1 agonist)enhanced apoptosis and inhibited proliferation in control cells.Conversely,treatment with NF449(a P2RX1 inhibitor)increased proliferation in both P2RX1-overexpressing and control SUP-B15 cells.Conclusion:Our findings indicate that P2RX1 may exert this function through modulating energy metabolism and calcium homeostasis,resulting in elevated intracellular calcium levels.Sustained elevation of calcium promotes apoptosis,whereas exogenous ATP activates P2RX1,enhances calcium influx,and attenuates the suppression of apoptosis associated with P2RX1 underexpression,ultimately correlating with improved treatment response.
