Objective To elucidate the regulatory mechanism of circRNAs in diabetic retinopathy.Methods Next-generation sequencing(NGS)was employed to identify circRNAs that are abnormally expressed in endothe-lial progenitor cel...Objective To elucidate the regulatory mechanism of circRNAs in diabetic retinopathy.Methods Next-generation sequencing(NGS)was employed to identify circRNAs that are abnormally expressed in endothe-lial progenitor cells(EPCs)under hyperglycemia(HG)conditions.The regulatory mechanism and predicted targets of this circRNA were also studied via bioinformatics analysis,luciferase reporter assays,angiogenic differentiation experiments,flow cytometry,and RT-qPCR.Results Circ-astrotactin 1(circ-Astn1)expression was decreased in EPCs under HG conditions,and circ-Astn1 overexpres-sion inhibited HG-induced endothelial damage.The miR-138-5p and silencing information regulator 2 related enzyme 1(SIRT1)were identified as circ-Astn1 downstream targets,which were further verified through luciferase reporter assays.SIRT1 silencing or miR-138-5p overexpression reversed the protective effect of circ-Astn1 on HG-induced endothelial cell dysfunction,as evidenced by increased apoptosis,abnormal vascular differentiation,and inflammatory factor secretion.SIRT1 overexpression reversed miR-138-5p-induced endothelial cell dysfunction under HG conditions.In vivo experiments confirmed that circ-Astnl overexpression promoted skin wound healing through the regulation of SIRT1.Conclusions These findings suggest that circ-Astn1 promotes SIRT1 expression by sponging miR-138-5p.Circ-Astn1 over-expression suppresses HG-induced endothelial cell damage via miR-138-5p/SIRT1 axis.展开更多
目的探讨乳腺癌中SIRT5的表达及其与PKM2及HK2相关性及临床意义。方法采用免疫组化SP法检测130例乳腺癌组织及癌旁正常组织中SIRT5、PKM2及HK2的表达,应用RT-PCR法检测60例乳腺癌组织及癌旁正常组织SIRT5、PKM2及HK2的基因表达,分析SIRT...目的探讨乳腺癌中SIRT5的表达及其与PKM2及HK2相关性及临床意义。方法采用免疫组化SP法检测130例乳腺癌组织及癌旁正常组织中SIRT5、PKM2及HK2的表达,应用RT-PCR法检测60例乳腺癌组织及癌旁正常组织SIRT5、PKM2及HK2的基因表达,分析SIRT5表达与乳腺癌临床病理特征的关系。结果乳腺癌组织中SIRT5、PKM2及HK2的平均光密度值均显著高于癌旁正常组织(5824.4±163.1 vs 2629.9±132.3、5980.3±232.2 vs 2164.9±121.3、6005.1±197.5 vs 2196.7±155.4,P<0.01),乳腺癌组织SIRT5、PKM2及HK2 mRNA的表达均高于癌旁正常组织(P<0.01)。乳腺癌组织中SIRT5与PKM2、HK2蛋白间呈正相关(r=0.647、r=0.600,P<0.01)。SIRT5阳性率与患者年龄、淋巴结转移、TNM分期、ER及PR表达差异无统计学意义(P=0.859,P=0.248,P=0.986,P=0.489,P=0.882);与肿瘤大小、组织学分级及HER-2表达差异有统计学意义(P=0.003,P=0.001,P=0.037)。结论SIRT5的过表达对判断乳腺癌的发生、发展及预后具有一定价值,并为SIRT5可能通过有氧糖酵解影响乳腺癌的发生、发展奠定基础。展开更多
基金supported by grants from the National Natural Science Foundation of China(No.81770483)Shanghai Tenth Hospital’s improvement plan for NSFC(No.04.03.17.070).
文摘Objective To elucidate the regulatory mechanism of circRNAs in diabetic retinopathy.Methods Next-generation sequencing(NGS)was employed to identify circRNAs that are abnormally expressed in endothe-lial progenitor cells(EPCs)under hyperglycemia(HG)conditions.The regulatory mechanism and predicted targets of this circRNA were also studied via bioinformatics analysis,luciferase reporter assays,angiogenic differentiation experiments,flow cytometry,and RT-qPCR.Results Circ-astrotactin 1(circ-Astn1)expression was decreased in EPCs under HG conditions,and circ-Astn1 overexpres-sion inhibited HG-induced endothelial damage.The miR-138-5p and silencing information regulator 2 related enzyme 1(SIRT1)were identified as circ-Astn1 downstream targets,which were further verified through luciferase reporter assays.SIRT1 silencing or miR-138-5p overexpression reversed the protective effect of circ-Astn1 on HG-induced endothelial cell dysfunction,as evidenced by increased apoptosis,abnormal vascular differentiation,and inflammatory factor secretion.SIRT1 overexpression reversed miR-138-5p-induced endothelial cell dysfunction under HG conditions.In vivo experiments confirmed that circ-Astnl overexpression promoted skin wound healing through the regulation of SIRT1.Conclusions These findings suggest that circ-Astn1 promotes SIRT1 expression by sponging miR-138-5p.Circ-Astn1 over-expression suppresses HG-induced endothelial cell damage via miR-138-5p/SIRT1 axis.
文摘目的探讨乳腺癌中SIRT5的表达及其与PKM2及HK2相关性及临床意义。方法采用免疫组化SP法检测130例乳腺癌组织及癌旁正常组织中SIRT5、PKM2及HK2的表达,应用RT-PCR法检测60例乳腺癌组织及癌旁正常组织SIRT5、PKM2及HK2的基因表达,分析SIRT5表达与乳腺癌临床病理特征的关系。结果乳腺癌组织中SIRT5、PKM2及HK2的平均光密度值均显著高于癌旁正常组织(5824.4±163.1 vs 2629.9±132.3、5980.3±232.2 vs 2164.9±121.3、6005.1±197.5 vs 2196.7±155.4,P<0.01),乳腺癌组织SIRT5、PKM2及HK2 mRNA的表达均高于癌旁正常组织(P<0.01)。乳腺癌组织中SIRT5与PKM2、HK2蛋白间呈正相关(r=0.647、r=0.600,P<0.01)。SIRT5阳性率与患者年龄、淋巴结转移、TNM分期、ER及PR表达差异无统计学意义(P=0.859,P=0.248,P=0.986,P=0.489,P=0.882);与肿瘤大小、组织学分级及HER-2表达差异有统计学意义(P=0.003,P=0.001,P=0.037)。结论SIRT5的过表达对判断乳腺癌的发生、发展及预后具有一定价值,并为SIRT5可能通过有氧糖酵解影响乳腺癌的发生、发展奠定基础。
文摘目的探究miRNA-424-5p(miR-424-5p)调控SIRT4表达影响胃癌细胞迁移和侵袭能力的潜在分子机制。方法利用RT-qPCR检测57例胃癌患者肿瘤组织与癌旁组织中miR-424-5p和SIRT4的表达水平。用脂质体法将miR-424-5p inhibitor和miR-424-5p mimic分别瞬时转染入MNK-28和HGC-27胃癌细胞中,RT-qPCR检测细胞中miR-424-5p和SIRT4 mRNA的表达水平,Western Blot检测细胞中SIRT4蛋白的表达量,划痕愈合实验和Transwell侵袭实验检测各组细胞的迁移和侵袭能力。双荧光素酶基因报告实验检测miR-424-5p对SIRT4的靶向调控机制。共转染miR-424-5p和SIRT4进一步验证miR-424-5p和SIRT4对胃癌细胞迁移和侵袭能力的影响。结果胃癌组织中miR-424-5p的表达明显高于癌旁组织(P<0.001),SIRT4的表达水平明显低于癌旁组织(P<0.001),相关性分析表明胃癌组织中miR-424-5p的表达与SIRT4呈负相关(r=-0.382,P=0.034)。此外,miR-424-5p高表达与肿瘤的浸润深度、TNM分期、脉管侵犯和神经侵犯显著相关(均P<0.05)。过表达miR-424-5p能促进胃癌细胞迁移和侵袭能力,而抑制miR-424-5p表达则呈现出相反的效果。过表达miR-424-5p可降低胃癌细胞中SIRT4 m RNA和蛋白的表达水平,相反,抑制miR-424-5p表达则上调胃癌细胞中SIRT4 m RNA和蛋白的表达水平。双荧光素酶报告基因实验显示miR-424-5p能显著影响野生型SIRT4-3’UTR表达载体的荧光素酶活性。回复实验结果显示miR-424-5p和SIRT4能显著影响胃癌细胞的迁移和侵袭能力。结论miR-424-5p在胃癌中高表达,其通过靶向抑制抑癌基因SIRT4的表达促进肿瘤细胞的迁移和侵袭,从而参与胃癌的的发生、发展。
