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pcDNANC组(转染pcDNA-NC空白载体)、pcDNA-SMAC组(转染pcDNA-SMAC载体)、siRNA-NC组(转染siRNANC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IC_(50)、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IC_(50)、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。
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 IC_(50),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 IC_(50),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,inhibit
作者
陈康
陈颖
牛宗新
康莉
祖里培亚·艾拜都拉
CHEN Kang;CHEN Ying;NIU Zongxin;KANG Li;ZULIPEYA·Aibaidula(Department of Thoracic Surgery,Xinjiang Uygur Autonomous Region People’s Hospital,Urumchi 830000,China)
出处
《肿瘤防治研究》
CAS
2023年第4期357-363,共7页
Cancer Research on Prevention and Treatment
基金
新疆维吾尔自治区自然科学基金(2017D01C136)。
关键词
caspase-3/Bcl-2/Bax
SMAC
肺腺癌
紫杉醇
增殖
侵袭
凋亡
caspase-3/Bcl-2/Bax signaling pathway
SMAC gene
Lung adenocarcinoma cells
Paclitaxel resistance sensitivity
Proliferation
Invasion
Apoptosis
