The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economic...The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economical and effective treatment methods.MicroRNA plays an important regulatory role in the occurrence and development of diseases.This study aims to explore the role and regulatory mechanism of miR⁃142a⁃3p in adriamycin(ADR)⁃induced renal tubular epithelial cell(TCMK⁃1)injury,with a focus on its potential as a therapeutic target for ADR nephropathy.First,cell viability was assessed using the CCK⁃8 kit,and a mouse renal tubular epithelial cell model induced by ADR was established.Subsequently,alterations in miR⁃142a⁃3p and its target gene ATG16L1 mRNA levels were quantified using RT⁃qPCR.Western blotting was used to detect the protein levels of autophagy marker proteins and pyroptosis marker proteins.Monodansylcadaverin(MDC)staining was performed and the autophagy of cells was detected by flow cytometry.The results showed that the relative expression of miR⁃142a⁃3p in TCMK⁃1 cells induced by ADR was increased and the relative expression of its target gene ATG16L1 was decreased(P<0.0001).Western blotting results showed that the levels of p62(P<0.001)and pyroptosis⁃related proteins(P<0.001)were increased,while the protein levels of autophagy⁃related proteins were decreased(P<0.05).The flow cytometry results showed that there was no difference in the mean fluorescence intensity of autoph⁃agosomes between the ADR group and the autophagosome inhibitor group(3⁃MA group)(P>0.05),indicating that after ADR induction,cell autophagy was inhibited and pyroptosis was enhanced.When the expression of miR⁃142a⁃3p was inhibited by transfecting miR⁃142a⁃3p inhibitor,the relative expression level of the target gene ATG16L1 was restored(P<0.001).Western blotting showed that the protein level of p62(P<0.01)and pyroptosis⁃related proteins(P<0.01)were decreased,and the protein level of autophagy⁃related proteins was restored(P<0.001).Flow cytometry results further indicated that cell autophagy was restored(P<0.0001).In conclusion,ADR targets ATG16L1 through miR⁃142a⁃3p to reduce the autophagy level of TCMK⁃1,and simultaneously activates GSDMD⁃mediated pyroptosis.展开更多
BACKGROUND The high mortality rate of hepatocellular carcinoma(HCC)in Egypt is due mainly to the increasing prevalence of hepatitis C virus infection(HCV)and late diagnosis of the carcinoma.MicroRNAs(miRNA),which regu...BACKGROUND The high mortality rate of hepatocellular carcinoma(HCC)in Egypt is due mainly to the increasing prevalence of hepatitis C virus infection(HCV)and late diagnosis of the carcinoma.MicroRNAs(miRNA),which regulate tumor proliferation and metastasis in HCC,may serve as a useful diagnostic approach for the early detection of HCC,thus decreasing its mortality.Meanwhile,endocan is a protein with angiogenic and inflammatory properties that are associated with tumor progression and poor outcomes.AIM To analyze the levels of miRNA 9-3p and endocan in HCV-infected HCC patients and correlate them with clinicopathological parameters.METHODS We compared levels of endocan and circulating miRNA 9-3p from 35 HCVrelated HCC patients to 33 patients with HCV-induced chronic liver disease and 32 age and gender matched healthy controls recruited from inpatient and outpatient clinics of the National Liver Institute,Menoufia University,Egypt in the period from January to March 2021 in a case-control study.Serum samples from all groups were analyzed for HCV.Endocan was measured by enzymelinked immunosorbent assays,and the expression levels of circulating miRNA 9-3p were measured by real-time quantitative reverse transcriptase PCR.RESULTS The levels of circulating miRNA 9-3p were significantly lower in the HCC group compared to the chronic liver disease(P<0.001)and control(P<0.001)groups,while levels in the chronic liver disease were significantly lower than those in the control group(P<0.001).The levels of serum endocan were significantly higher in the HCC group compared to the chronic liver disease(P<0.001)and control(P<0.001)groups.Moreover miRNA 9-3p and endocan performed better thanα-fetoprotein in discriminating HCC patients from cirrhosis and healthy patients.The levels of miRNA 9-3p were significantly inversely correlated to vascular invasion(P=0.002),stage of advancement of Barcelona Clinical Liver Cancer(P<0.001)and the metastatic site(P<0.001)of the HCC group.CONCLUSION Circulating miRNA 9-3p and endocan can be used as novel biomarkers for the early diagnosis of HCV-related HCC.展开更多
OBJECTIVE MicroR NA(miR NA)holds promise as a novel therapeutic tool for cancer treatment.However,the transfection efficiency of current delivery systems represents a bottleneck for clinical applications.Here,we demon...OBJECTIVE MicroR NA(miR NA)holds promise as a novel therapeutic tool for cancer treatment.However,the transfection efficiency of current delivery systems represents a bottleneck for clinical applications.Here,we demonstrate that gap junctions mediate an augmentative effect on the antiproliferation mediated by mi R-124-3p in U87 and C6 glioblastoma cells.METHODS The functional inhibition of gap junctions using either si RNA or pharmacological inhibition eliminated the mi R-124-3p-mediated antiproliferation,whereas the enhancement of gap junctions with retinoic acid treatment augmented this mi R-124-3p-mediated antiproliferation.A similar effect was observed in glioblastoma xenograft models.RESULTS More importantly,patch clamp and co-culture assays demonstrated the transmission of mi R-124-3p through gap junction channels into adjacent cells.In further exploring the impact of gap junction-mediated transport of mi R-124-3p on mi R-124-3p target pathways,we found that mi R-124-3p inhibited glioblastoma cell growth in part by decreasing the protein expression of cyclindependent kinase 6,leading to cel cycle arrest at the G0/G1phase;moreover,pharmacological regulation of gap junctions affected this cell cycle arrest.CONCLUSION Our results indicate that the″bystander″effects of functional gap junctions composed of connexin 43 enhance the antitumor effect of mi R-124-3p in glioblastoma cells by transferring mi R-124-3p to adjacent cells,thereby enhancing G0/G1cell cycle arrest.These observations provide a new guiding strategy for the clinical application of mi RNA therapy in tumor treatment.展开更多
目的：探讨miRNA-34c在人脑胶质瘤组织中的表达情况及临床意义。方法应用定量PCR方法对18例WHO分级Ⅱ～Ⅳ级的甲醛固定石蜡包埋的脑胶质瘤标本和外伤或脑出血患者内减压手术获得的5例脑组织标本分别进行miR-34c-3p和miR-34c-5p含量的检...目的：探讨miRNA-34c在人脑胶质瘤组织中的表达情况及临床意义。方法应用定量PCR方法对18例WHO分级Ⅱ～Ⅳ级的甲醛固定石蜡包埋的脑胶质瘤标本和外伤或脑出血患者内减压手术获得的5例脑组织标本分别进行miR-34c-3p和miR-34c-5p含量的检测，比较不同级别胶质瘤和正常脑组织中miR-34c-3p和miR-34c-5p的表达差异，分析miR-34c-3p和miR-34c-5p的表达和胶质瘤恶性程度的相关性。结果与正常脑组织相比，胶质瘤中miR-34c-3p和miR-34c-5p的表达皆显著减少（ P ＜0＊．05），而且miR-34c-3p和miR-34c-5p的表达随着胶质瘤级别或恶性程度的增加分别减少，呈显著的负相关（ miR-34c-3p的spearman相关系数＝－0．856， P ＜0．01；miR-34c-5p的spearman相关系数＝－0．767， P ＜0．01）。结论 MiR-34c在人脑胶质瘤细胞中的表达显著降低，而且表达随着肿瘤分级或恶性程度的增高而减少，与胶质瘤级别的升高呈显著的负相关。 MiR-34c在胶质瘤的进展中有重要的意义，可作为脑胶质瘤临床分级的重要指标。展开更多
文摘The incidence rate of kidney diseases in China has always remained high.At present,the clinical treatment mainly focuses on symptomatic treatment to delay the progression of the disease,and there is a lack of economical and effective treatment methods.MicroRNA plays an important regulatory role in the occurrence and development of diseases.This study aims to explore the role and regulatory mechanism of miR⁃142a⁃3p in adriamycin(ADR)⁃induced renal tubular epithelial cell(TCMK⁃1)injury,with a focus on its potential as a therapeutic target for ADR nephropathy.First,cell viability was assessed using the CCK⁃8 kit,and a mouse renal tubular epithelial cell model induced by ADR was established.Subsequently,alterations in miR⁃142a⁃3p and its target gene ATG16L1 mRNA levels were quantified using RT⁃qPCR.Western blotting was used to detect the protein levels of autophagy marker proteins and pyroptosis marker proteins.Monodansylcadaverin(MDC)staining was performed and the autophagy of cells was detected by flow cytometry.The results showed that the relative expression of miR⁃142a⁃3p in TCMK⁃1 cells induced by ADR was increased and the relative expression of its target gene ATG16L1 was decreased(P<0.0001).Western blotting results showed that the levels of p62(P<0.001)and pyroptosis⁃related proteins(P<0.001)were increased,while the protein levels of autophagy⁃related proteins were decreased(P<0.05).The flow cytometry results showed that there was no difference in the mean fluorescence intensity of autoph⁃agosomes between the ADR group and the autophagosome inhibitor group(3⁃MA group)(P>0.05),indicating that after ADR induction,cell autophagy was inhibited and pyroptosis was enhanced.When the expression of miR⁃142a⁃3p was inhibited by transfecting miR⁃142a⁃3p inhibitor,the relative expression level of the target gene ATG16L1 was restored(P<0.001).Western blotting showed that the protein level of p62(P<0.01)and pyroptosis⁃related proteins(P<0.01)were decreased,and the protein level of autophagy⁃related proteins was restored(P<0.001).Flow cytometry results further indicated that cell autophagy was restored(P<0.0001).In conclusion,ADR targets ATG16L1 through miR⁃142a⁃3p to reduce the autophagy level of TCMK⁃1,and simultaneously activates GSDMD⁃mediated pyroptosis.
文摘BACKGROUND The high mortality rate of hepatocellular carcinoma(HCC)in Egypt is due mainly to the increasing prevalence of hepatitis C virus infection(HCV)and late diagnosis of the carcinoma.MicroRNAs(miRNA),which regulate tumor proliferation and metastasis in HCC,may serve as a useful diagnostic approach for the early detection of HCC,thus decreasing its mortality.Meanwhile,endocan is a protein with angiogenic and inflammatory properties that are associated with tumor progression and poor outcomes.AIM To analyze the levels of miRNA 9-3p and endocan in HCV-infected HCC patients and correlate them with clinicopathological parameters.METHODS We compared levels of endocan and circulating miRNA 9-3p from 35 HCVrelated HCC patients to 33 patients with HCV-induced chronic liver disease and 32 age and gender matched healthy controls recruited from inpatient and outpatient clinics of the National Liver Institute,Menoufia University,Egypt in the period from January to March 2021 in a case-control study.Serum samples from all groups were analyzed for HCV.Endocan was measured by enzymelinked immunosorbent assays,and the expression levels of circulating miRNA 9-3p were measured by real-time quantitative reverse transcriptase PCR.RESULTS The levels of circulating miRNA 9-3p were significantly lower in the HCC group compared to the chronic liver disease(P<0.001)and control(P<0.001)groups,while levels in the chronic liver disease were significantly lower than those in the control group(P<0.001).The levels of serum endocan were significantly higher in the HCC group compared to the chronic liver disease(P<0.001)and control(P<0.001)groups.Moreover miRNA 9-3p and endocan performed better thanα-fetoprotein in discriminating HCC patients from cirrhosis and healthy patients.The levels of miRNA 9-3p were significantly inversely correlated to vascular invasion(P=0.002),stage of advancement of Barcelona Clinical Liver Cancer(P<0.001)and the metastatic site(P<0.001)of the HCC group.CONCLUSION Circulating miRNA 9-3p and endocan can be used as novel biomarkers for the early diagnosis of HCV-related HCC.
基金The project supported by National Natural Science Foundation of China(81473234,U1303221)
文摘OBJECTIVE MicroR NA(miR NA)holds promise as a novel therapeutic tool for cancer treatment.However,the transfection efficiency of current delivery systems represents a bottleneck for clinical applications.Here,we demonstrate that gap junctions mediate an augmentative effect on the antiproliferation mediated by mi R-124-3p in U87 and C6 glioblastoma cells.METHODS The functional inhibition of gap junctions using either si RNA or pharmacological inhibition eliminated the mi R-124-3p-mediated antiproliferation,whereas the enhancement of gap junctions with retinoic acid treatment augmented this mi R-124-3p-mediated antiproliferation.A similar effect was observed in glioblastoma xenograft models.RESULTS More importantly,patch clamp and co-culture assays demonstrated the transmission of mi R-124-3p through gap junction channels into adjacent cells.In further exploring the impact of gap junction-mediated transport of mi R-124-3p on mi R-124-3p target pathways,we found that mi R-124-3p inhibited glioblastoma cell growth in part by decreasing the protein expression of cyclindependent kinase 6,leading to cel cycle arrest at the G0/G1phase;moreover,pharmacological regulation of gap junctions affected this cell cycle arrest.CONCLUSION Our results indicate that the″bystander″effects of functional gap junctions composed of connexin 43 enhance the antitumor effect of mi R-124-3p in glioblastoma cells by transferring mi R-124-3p to adjacent cells,thereby enhancing G0/G1cell cycle arrest.These observations provide a new guiding strategy for the clinical application of mi RNA therapy in tumor treatment.
文摘目的：探讨miRNA-34c在人脑胶质瘤组织中的表达情况及临床意义。方法应用定量PCR方法对18例WHO分级Ⅱ～Ⅳ级的甲醛固定石蜡包埋的脑胶质瘤标本和外伤或脑出血患者内减压手术获得的5例脑组织标本分别进行miR-34c-3p和miR-34c-5p含量的检测，比较不同级别胶质瘤和正常脑组织中miR-34c-3p和miR-34c-5p的表达差异，分析miR-34c-3p和miR-34c-5p的表达和胶质瘤恶性程度的相关性。结果与正常脑组织相比，胶质瘤中miR-34c-3p和miR-34c-5p的表达皆显著减少（ P ＜0＊．05），而且miR-34c-3p和miR-34c-5p的表达随着胶质瘤级别或恶性程度的增加分别减少，呈显著的负相关（ miR-34c-3p的spearman相关系数＝－0．856， P ＜0．01；miR-34c-5p的spearman相关系数＝－0．767， P ＜0．01）。结论 MiR-34c在人脑胶质瘤细胞中的表达显著降低，而且表达随着肿瘤分级或恶性程度的增高而减少，与胶质瘤级别的升高呈显著的负相关。 MiR-34c在胶质瘤的进展中有重要的意义，可作为脑胶质瘤临床分级的重要指标。
