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陈康, 陈颖, 牛宗新, 康莉, 祖里培亚·艾拜都拉. 基于caspase-3/Bcl-2/Bax信号通路探究SMAC基因对肺腺癌细胞紫杉醇敏感度及细胞活性的影响[J]. 肿瘤防治研究, 2023, 50(4): 357-363. DOI: 10.3971/j.issn.1000-8578.2023.22.1114
引用本文: 陈康, 陈颖, 牛宗新, 康莉, 祖里培亚·艾拜都拉. 基于caspase-3/Bcl-2/Bax信号通路探究SMAC基因对肺腺癌细胞紫杉醇敏感度及细胞活性的影响[J]. 肿瘤防治研究, 2023, 50(4): 357-363. DOI: 10.3971/j.issn.1000-8578.2023.22.1114
CHEN Kang, CHEN Ying, NIU Zongxin, KANG Li, ZULIPEYA·Aibaidula. Effect of SMAC Gene on Sensitivity of Lung Adenocarcinoma Cells to Paclitaxel and Cell Viability Based on caspase-3/Bcl-2/Bax Signaling Pathway[J]. Cancer Research on Prevention and Treatment, 2023, 50(4): 357-363. DOI: 10.3971/j.issn.1000-8578.2023.22.1114
Citation: CHEN Kang, CHEN Ying, NIU Zongxin, KANG Li, ZULIPEYA·Aibaidula. Effect of SMAC Gene on Sensitivity of Lung Adenocarcinoma Cells to Paclitaxel and Cell Viability Based on caspase-3/Bcl-2/Bax Signaling Pathway[J]. Cancer Research on Prevention and Treatment, 2023, 50(4): 357-363. DOI: 10.3971/j.issn.1000-8578.2023.22.1114

基于caspase-3/Bcl-2/Bax信号通路探究SMAC基因对肺腺癌细胞紫杉醇敏感度及细胞活性的影响

Effect of SMAC Gene on Sensitivity of Lung Adenocarcinoma Cells to Paclitaxel and Cell Viability Based on caspase-3/Bcl-2/Bax Signaling Pathway

  • 摘要:
    目的 基于caspase-3/Bcl2/Bax信号通路探究SMAC基因对肺腺癌细胞紫杉醇敏感度及细胞活性的影响。
    方法 建立肺腺癌紫杉醇耐药细胞株A549/Taxol,将细胞分为pcDNA-NC组(转染pcDNA-NC空白载体)、pcDNA-SMAC组(转染pcDNA-SMAC载体)、siRNA-NC组(转染siRNA-NC空病毒载体)和siRNA-SMAC组(转染siRNA-SMAC慢病毒载体)。qRT-PCR法检测细胞中SMAC mRNA表达;MTT法检测细胞敏感度;克隆实验法检测细胞增殖能力;Transwell法检测细胞侵袭能力;流式细胞术检测细胞凋亡能力;Western blot法检测细胞中caspase-3、Bcl-2和Bax蛋白表达。
    结果 肺腺癌A549细胞较BEAS-2B正常细胞中SMAC mRNA表达明显降低(P < 0.05)。pcDNA-SMAC组较pcDNA-NC组细胞中SMAC mRNA表达显著升高(P < 0.05)。和siRNA-NC组相比,siRNA-SMAC组细胞中SMAC mRNA表达显著降低(P < 0.05)。和pcDNA-NC组相比,pcDNA-SMAC组细胞IC50、细胞克隆数、细胞侵袭能力及Bcl-2蛋白和Bcl-2/Bax比值均显著降低,细胞耐药指数逆转倍数为2.51倍,细胞凋亡能力及caspase-3和Bax蛋白表达明显高于pcDNA-NC组(P < 0.05)。和siRNA-NC组相比,siRNA-SMAC组细胞IC50、细胞克隆数、细胞侵袭能力及Bcl-2蛋白和Bcl-2/Bax比值均显著升高,细胞凋亡能力及caspase-3和Bax蛋白表达明显降低(P < 0.05)。
    结论 高表达SMAC可增加肺腺癌细胞的紫杉醇敏感度、抑制细胞增长和侵袭、促进细胞凋亡,且对caspase-3/Bcl-2/Bax信号通路有一定调控作用。

     

    Abstract:
    Objective To investigate the effect of the SMAC gene on paclitaxel sensitivity and cellular activity in lung adenocarcinoma cells based on the caspase-3/Bcl-2/Bax signaling pathway.
    Methods A paclitaxel-resistant cell line A549/Taxol was established for lung adenocarcinoma, and the cells were divided into four following groups: pcDNA-NC (transfected with pcDNA-NC blank vector), pcDNA-SMAC (transfected with pcDNA-SMAC vector), siRNA-NC (transfected with siRNA-NC empty virus vector), and siRNA-SMAC groups (transfected with siRNA-SMAC lentiviral vector). The SMAC mRNA expression in cells was detected by qRT-PCR; cell sensitivity was detected by MTT; cell proliferation ability was detected by cloning assay; cell invasion ability was detected by Transwell; apoptosis ability was detected by flow cytometry assay; and caspase-3, Bcl-2 and Bax protein expression in cells were detected by Western blot analysis.
    Results The SMAC mRNA expression was significantly lower in A549 cells compared with BEAS-2B cells (P < 0.05). The SMAC mRNA expression was significantly higher in the pcDNA-SMAC group than that in the pcDNA-NC group cells (P < 0.05). The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than that in the siRNA-NC group. The SMAC mRNA expression was significantly lower in the cells of the siRNA-SMAC group (P < 0.05) than in the siRNA-NC group. Compared with the pcDNA-NC group, the cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly lower in the pcDNA-SMAC group, the cell resistance index reversal was 2.51-fold, and the apoptosis ability and caspase-3, as well as Bax protein expression, were significantly higher (P < 0.05). Compared with the siRNA-NC group, cell IC50, cell clone number, cell invasion ability, and Bcl-2 protein and Bcl-2/Bax ratio were significantly higher in the siRNA-SMAC group, and apoptosis ability and caspase-3 and Bax protein expression were significantly lower (P < 0.05).
    Conclusion High expression of SMAC increases paclitaxel sensitivity, inhibits cell growth and invasion, promotes apoptosis in lung adenocarcinoma cells, and has a regulatory effect on the caspase-3/Bcl-2/Bax signaling pathway.

     

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