摘要
目的探讨基质细胞衍生因子-1(SDF-1)过表达对人胃癌耐药细胞株SGC7901/L-OHP增殖、凋亡、耐药的影响。方法通过脂质体将真核表达载体pcDNA3.1-SDF-1转染至人胃癌耐药细胞株SGC7901/L-OHP中,反转录聚合酶链反应(RT-PCR)及In-cell Western验证转染效率。噻唑蓝(MTT)、流式细胞术检测SDF-1过表达对人胃癌耐药细胞株SGC7901/L-OHP增殖、细胞周期、凋亡及奥沙利铂耐药的影响。In-cell Western检测SDF-1过表达对survivin、Bcl-xl、cyclinD1、caspase-3蛋白表达的影响。结果在pcDNA3.1-SDF-1转染后的SGC7901/L-OHP细胞中,SDF-1 mRNA和蛋白表达显著增加(均P<0.01)。与空白对照组及阴性对照组相比,SDF-1过表达的人胃癌耐药细胞株SGC7901/L-OHP细胞增殖加速,其G 0/G 1期细胞减少、G 2/M期细胞增多,细胞凋亡率降低,且细胞对奥沙利铂的耐药性明显增加。SDF-1过表达能够明显上调survivin、Bcl-xl、cyclinD1蛋白表达水平,同时下调caspase-3蛋白表达水平。结论SDF-1过表达能够促进胃癌SGC7901/L-OHP细胞增殖,抑制其凋亡,提高其对奥沙利铂的耐药性,机制可能与上调凋亡抑制基因survivin、Bcl-xl、cyclinD1的表达,同时下调凋亡促进基因caspase-3的表达有关。
Objective To investigate the effects of stromal cell-derived factor-1(SDF-1)overexpression on proliferation,apoptosis and drug resistance of human gastric cancer drug-resistant cell line SGC7901/L-OHP.Methods The eukaryotic expression vector pcDNA3.1-SDF-1 was transfected into human gastric cancer drug-resistant cell line SGC7901/L-OHP by liposome,and the transfection efficiency was verified by RT-PCR and In-cell Western.MTT and flow cytometry were used to detect the effects of SDF-1 overexpression on proliferation,apoptosis and cell cycle of human gastric cancer drug-resistant cell line SGC7901/L-OHP.MTT assay was used to detect the effect of SDF-1 overexpression on the resistance of oxaliplatin.In-cell Western assay was used to detect the effects of SDF-1 overexpression on the protein expression of survivin,Bcl-xl,cyclinD1 and caspase-3.Results The mRNA and protein expression of SDF-1 in SGC7901/L-OHP cells transfected with pcDNA3.1-SDF-1 were increased significantly.Compared with the blank and negative control groups,overexpression of SDF-1 could significantly accelerate the proliferation of SGC7901/L-OHP cells,reduced G 0/G 1 cells,increased G 2/M cells and decreased apoptotic rate,moreover increased the resistance to oxaliplatin.SDF-1 overexpression could significantly up-regulate the protein expression levels of survivin,Bcl-xl and cyclinD1,while down-regulate the protein expression level of caspase-3 protein.Conclusion Overexpression of SDF-1 can promote the proliferation of SGC7901/L-OHP cells,inhibit their apoptosis,and improve the resistance of gastric cancer cells to oxaliplatin.The mechanism may be related to up-regulate the expression of survivin,Bcl-xl and cyclinD1,meanwhile down-regulate the expression of caspase-3.
作者
刘小慧
贺曼曼
张伟
李静
郭明海
冯运章
赵国杰
赵飞
LIU Xiaohui;HE Manman;ZHANG Wei;LI Jing;GUO Minghai;FENG Yunzhang;ZHAO Guojie;ZHAO Fei(Second Department of General Surgery;Department of Anesthesiology,Handan Municipal Central Hospital,Handan,Hebei 056001,China)
出处
《重庆医学》
CAS
2020年第5期695-700,共6页
Chongqing medicine
基金
河北省重点研发计划自筹项目（172777201）
