摘要
目的 探讨自噬基因Beclin-1过表达对A549肺癌细胞自噬和增殖的影响以及其作用机制.方法 用实时荧光定量聚合酶链反应(FQ-PCR)和Western blot的方法分别检测转染后各组A549肺癌细胞Beclin-1 mRNA和蛋白的表达;噻唑蓝(MTT)法检测各组A549细胞生长增殖;单丹磺酰戊二胺(MDC)法检测各组A549细胞自噬的变化;Western blot方法检测各组A549细胞食管癌相关基因4(ECRG4)蛋白表达变化.结果 与对照组比较,感染pLenex-Beclin-1慢病毒颗粒后,A549细胞中Beclin-1 mRNA和蛋白表达显著增加(P<0.05);与对照组比较,感染pLenex-Beclin-1慢病毒颗粒后,Beclin-1过表达组细胞群体平均倍增时间[(46.0±2.4)h]显著延长(P<0.05);与对照组比较,感染pLenex-Beclin-1慢病毒颗粒后,自噬细胞阳性数量[(10.0±1.5)个]显著增加(P<0.05);与对照组比较,感染pLenex-Beclin-1慢病毒颗粒后,ECRG4蛋白表达增加(P<0.05).结论 自噬基因Beclin-1通过ECRG4通路抑制A549肺癌细胞增殖.
Objective To investigate the effect and molecular mechanism of autophagy-related gene Beclin-1 overexpression on autophagy and proliferation of A549 lung cancer cells.Methods Fluorescence quantitative real-time fluorescent quantitative polymerase chain reaction (FQ-PCR) and Western blotting were used to detect beclin1 mRNA and protein expression levels of transfected cells in each group.The growth and proliferation of A549 lung cancer cells were observed by methyl thiazol tetrazolium (MTT) method.Monodansylcadaverine (MDC) method was used to detect autophagy.Western blotting was used to detect esophageal cancer related gene 4 (ECRG4) protein expression of transfected cells in each group.Results After A549 cells were infected with pLenex-Beclin-1 lentiviral particles,the expression levels of Beclin-1 mRNA and protein were statistically increased (P 〈 0.05),the population doubling time of A549 ceils [(46.0 ± 2.4) h] was significantly prolonged (P 〈 0.05),the autophagy was increased,the number of autophagy positive cells (10.0 ± 1.5) was increased (P 〈 0.05),and the ECRG4 expression was significantly up-regulated (P 〈 0.05).Conclusion Autophagy-related gene Beclin-1 inhibited A549 lung cancer cells proliferation by ECRG4 pathway.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2016年第4期1005-1007,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(U1304817)
郑州市科技攻关计划资助项目(141PPTGHG298)
关键词
BECLIN-1
自噬
增殖
食管癌
食管癌相关基因4
Beclin -1
Autophagy
Proliferaion
Esophageal cancer
Esophageal cancer related gene 4