集缩素NCAPG对非小细胞肺癌增殖和凋亡的影响及其可能机制

The eEffect and mechanism of non-SMC chromosome-associated polypeptide G on the proliferation and apoptosis of non-small cell lung cancer

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.