摘要
目的探讨非小细胞肺癌细胞中集缩素NCAPG表达对A549与H1299细胞增殖、凋亡的影响及其机制。方法构建shRNA-NCAPG慢病毒载体转染A549及NCI-H1299细胞,实时定量PCR法(qRT-PCR)及免疫印迹法(Western blot)验证两种细胞中NCAPG表达情况。应用CCK-8方法检测对照组及下调组的细胞增殖差异,明确NCAPG表达对细胞增殖的抑制作用。Annexin V/PI双染法流式细胞术检测两组细胞中早期凋亡及晚期凋亡细胞比例。Western blot法比较两组细胞中Caspase-3、Bax及Bcl-2的表达水平。结果 shRNA-NCAPG转染组细胞的NCAPG表达量低于对照组。下调NCAPG表达抑制A549及NCI-H1299细胞增殖,增加早期凋亡及晚期细胞凋亡比例,增加肺癌细胞促凋亡相关蛋白Caspase-3及Bax的表达、抑制Bcl-2蛋白表达。结论下调NCAPG表达诱导非小细胞肺癌A549及H1299细胞增殖,通过线粒体通路抑制其细胞凋亡。
Objective To investigate the effect of non-SMC chromosome-associated polypeptide G(NCAPG) on A549 and NCI-H1299 cell proliferation and apoptosis and its possible mechanism. Methods The sh-NCAPG plasmid was constructed and transfected into A549 and NCI-H1299 cells. The expression of NCAPG was verified by qRT-PCR and Western blot. CCK-8 method was used to detect the cell proliferation. The cell apoptosis was stained with Annexin V/PI, and the proportion of early apoptotic cells and late apoptotic cells was detected by flow cytometry. The expressions of Caspase-3, Bax and Bcl-2 were determined by Western blot. Results NCAPG expression level was down-regulated in sh-NCAPG group. The down-regulation of NCAPG expression levle inhibited the proliferation of A549 and NCI-H1299 cells, increased the proportion of both early and late apoptotic cells, upregulated the expression levels of pro-apoptotic proteins Caspase-3 and Bax, but inhibited anti-apoptotic protein Bcl-2 expression level. Conclusion Silencing NCAPG inhibits cells proliferation and induces apoptosis via mitochondrial pathway in non-small cell lung cancer.
作者
刘宇
李继佳
刘长浩
王阁兵
张晨雷
于占武
余平文
刘宏旭
LIU Yu;LI Ji-jia;LIU Chang-hao;WANG Ge-bing;ZHANG Chen-lei;YU Zhan-wu;YU Ping-wen;LIU Hong-xu(First Ward of Thoracic Surgery,Cancer Hospital of China Medical University,Liaoning Cancer Hospital&Institute,Shenyang 110042,China)
出处
《解剖科学进展》
CAS
2021年第3期312-315,共4页
Progress of Anatomical Sciences
基金
辽宁省自然科学基金(2015020258)。
关键词
集缩素NCAPG
非小细胞肺癌
细胞增殖
细胞凋亡
non-SMC chromosome-associated polypeptide G
non-small cell lung cancer
proliferation
apoptosis