目的探讨自噬相关基因微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)和自噬相关基因-5(autophagy related gene-5,ATG5)在胃癌细胞发生发展不同阶段的表达情况并分析其临床意义。方法选取胃黏膜正常细胞、...目的探讨自噬相关基因微管相关蛋白1轻链3(microtubule-associated protein 1 light chain 3,LC3)和自噬相关基因-5(autophagy related gene-5,ATG5)在胃癌细胞发生发展不同阶段的表达情况并分析其临床意义。方法选取胃黏膜正常细胞、胃黏膜不典型增生细胞、胃癌细胞共58例,分别记为正常组(17例)、不典型组(19例)、胃癌组(22例),采用qRT-PCR、Western Blot、免疫组织化学法检测每组中自噬相关基因LC3和ATG5的mRNA及蛋白表达水平,应用SPSS 23.0软件分析正常组、不典型组、胃癌组3组间LC3和ATG5的mRNA及蛋白表达情况,并分析LC3和ATG5蛋白表达的相关性以及LC3和ATG5蛋白表达与胃癌患者临床病理学参数的关系。结果正常组、不典型组、胃癌组中LC3和ATG5 mRNA及蛋白表达水平逐渐升高,各组间差异均有统计学意义(P<0.05)。LC3蛋白表达情况与性别、年龄、浸润深度、分化程度及淋巴结状态均不存在相关性。ATG5蛋白表达情况与浸润深度、分化程度及淋巴结状态均有相关性。LC3和ATG5蛋白在3组中均呈正相关(P<0.05)。结论细胞自噬可促进胃癌的发生发展,其机制可能与LC3、ATG5过度表达有关。LC3、ATG5在促进胃癌的发生发展中起相互协同的作用。展开更多
目的:探究miR-30a通过靶向调控Beclin-1、ATG5表达对食管癌细胞自噬及增殖过程的影响。方法:靶基因预测、双荧光素酶报告实验验证miR-30a对Beclin-1、ATG5的靶向调控作用。GEO、Human Protein Atlas和GEPIA在线分析TCGA数据库和GTEx项...目的:探究miR-30a通过靶向调控Beclin-1、ATG5表达对食管癌细胞自噬及增殖过程的影响。方法:靶基因预测、双荧光素酶报告实验验证miR-30a对Beclin-1、ATG5的靶向调控作用。GEO、Human Protein Atlas和GEPIA在线分析TCGA数据库和GTEx项目中食管癌组织及癌旁组织miR-30a、Beclin-1、ATG5表达、患者预后生存期。免疫组化检测45例食管癌及癌旁组织标本Beclin-1、ATG5表达,并分析Beclin-1、ATG5表达与患者临床病理参数的关系。采用脂质体LipofectamineTM3000将靶向Beclin-1、ATG5的miR-30a-mimic转染至Eca-109细胞,体外常规培养并分为3组:未转染组(control组)、转染无义RNA的阴性对照组(mimic NC组)、转染miR-30a-mimic的干扰组(miR-30a组)。MTT和克隆形成实验测定细胞增殖活力;qRT-PCR检测miR-30a、Beclin-1、ATG5 mRNA表达;划痕实验检测细胞迁移能力;Transwell小室实验检测细胞迁移和侵袭;Western blot检测Beclin-1、ATG5、LC3Ⅱ、p62蛋白表达。透射电镜观察细胞自噬泡形成情况。结果:生信分析结果显示,食管癌组织中miR-30a表达高于正常组织,Beclin-1、ATG5表达低于正常组织,且预后生存期与miR-30a表达呈负相关,与Beclin-1、ATG5表达呈正相关(P<0.05)。与control组和mimic NC组相比,miR-30a组细胞增殖、迁移和侵袭能力、p62蛋白表达明显升高,Beclin-1、ATG5、LC3Ⅱ表达、细胞自噬活力明显降低(P<0.05)。而control组与mimic NC组上述指标差异无统计学意义(P>0.05)。结论:miR-30a靶向调控Beclin-1、ATG5表达;Beclin-1、ATG5在食管癌组织和细胞中表达均降低,具有肿瘤抑制基因作用,可能通过抑制癌细胞增殖、迁移和侵袭、促进自噬发挥抑癌作用,为通过靶向调控自噬途径治疗食管癌提供了新思路。展开更多
肝脏疾病作为危害人类健康的常见病和多发性疾病,在国内外对其的防治与治疗仍是个严峻的问题。细胞自噬是细胞体内自我调节的重要方式,细胞生长、分化、存活和自我平衡都依赖于此,同时在对抗慢性肝炎病毒感染、酒精性肝病、脂肪肝等肝...肝脏疾病作为危害人类健康的常见病和多发性疾病,在国内外对其的防治与治疗仍是个严峻的问题。细胞自噬是细胞体内自我调节的重要方式,细胞生长、分化、存活和自我平衡都依赖于此,同时在对抗慢性肝炎病毒感染、酒精性肝病、脂肪肝等肝脏疾病时发挥重要作用。研究表明,细胞自噬受到自噬相关基因-5(autophagy related gene-5,Atg5)的调控,通过Atg5调控细胞自噬来对抗肝脏疾病也引起越来越多的关注。例如在肝细胞中,上调Atg5的表达量可促进细胞自噬,从而减少内质网应激(ER)介导的损伤,这是治疗脂肪性肝炎的一个新策略。本文就Atg5通过调控自噬参与常见肝脏疾病的内在机制做一总结。展开更多
Endothelial-mesenchymal transition(EndoMT)is associated with damage to blood-brain barrier(BBB)integrity.Circular RNAs(circRNAs)are highly expressed in the brain and are involved in brain diseases;however,whether circ...Endothelial-mesenchymal transition(EndoMT)is associated with damage to blood-brain barrier(BBB)integrity.Circular RNAs(circRNAs)are highly expressed in the brain and are involved in brain diseases;however,whether circR NAs regulate the EndoM T in the brain remains unknown.Our study demonstrated that circH ECW2 regulated the EndoM T by directly binding to MIR30 D,a significantly downregulated miR NA from miR NA profiling,which subsequently caused an increased expression of ATG5.These findings shed new light on the understanding of the noncanonical role of ATG5 in the EndoMT induced by methamphetamine or lipopolysaccharide.The in vivo relevance was confirmed as microinjection of circHecw2 siRNA lentivirus into the mouse hippocampus suppressed the EndoMT induced by LPS.These findings provide novel insights regarding the contribution of circHECW2 to the nonautophagic role of ATG5 in the EndoMT process in the context of drug abuse and the broad range of neuroinflammatory disorders.展开更多
文摘肝脏疾病作为危害人类健康的常见病和多发性疾病,在国内外对其的防治与治疗仍是个严峻的问题。细胞自噬是细胞体内自我调节的重要方式,细胞生长、分化、存活和自我平衡都依赖于此,同时在对抗慢性肝炎病毒感染、酒精性肝病、脂肪肝等肝脏疾病时发挥重要作用。研究表明,细胞自噬受到自噬相关基因-5(autophagy related gene-5,Atg5)的调控,通过Atg5调控细胞自噬来对抗肝脏疾病也引起越来越多的关注。例如在肝细胞中,上调Atg5的表达量可促进细胞自噬,从而减少内质网应激(ER)介导的损伤,这是治疗脂肪性肝炎的一个新策略。本文就Atg5通过调控自噬参与常见肝脏疾病的内在机制做一总结。
文摘Endothelial-mesenchymal transition(EndoMT)is associated with damage to blood-brain barrier(BBB)integrity.Circular RNAs(circRNAs)are highly expressed in the brain and are involved in brain diseases;however,whether circR NAs regulate the EndoM T in the brain remains unknown.Our study demonstrated that circH ECW2 regulated the EndoM T by directly binding to MIR30 D,a significantly downregulated miR NA from miR NA profiling,which subsequently caused an increased expression of ATG5.These findings shed new light on the understanding of the noncanonical role of ATG5 in the EndoMT induced by methamphetamine or lipopolysaccharide.The in vivo relevance was confirmed as microinjection of circHecw2 siRNA lentivirus into the mouse hippocampus suppressed the EndoMT induced by LPS.These findings provide novel insights regarding the contribution of circHECW2 to the nonautophagic role of ATG5 in the EndoMT process in the context of drug abuse and the broad range of neuroinflammatory disorders.
