基质细胞衍生因子-1基因转染对人胃癌细胞株SGC7901增殖、耐药的影响及其机制  

Effects of SDF-1 gene transfection on proliferation and oxaliplatin resistance of gastric cancer SGC7901 cells

作　　者：刘小慧 贺曼曼 张伟[1] 李静[1] 牛广旭[1] 冯运章[1] LIU Xiaohui;HE Manman;ZHANG Wei;LI Jing;NIU Guangxu;FENG Yunzhang(Handan Central Hospital,Handan 056001,China)

机构地区：[1]邯郸市中心医院

出　　处：《山东医药》2019年第18期23-27,共5页Shandong Medical Journal

基　　金：河北省重点研发计划自筹项目(172777201)

摘　　要：目的观察基质细胞衍生因子-1(SDF-1)基因转染对人胃癌细胞株SGC7901增殖、耐药的影响,并探讨其可能作用机制。方法 取对数生长期 SGC7901 细胞分为实验组、阴性对照组、空白对照组,实验组、阴性对照组分别转染pcDNA3.1-SDF-1质粒、pcDNA3.1质粒(空白质粒),空白对照组不做任何处理。质粒转染48 h时分别采用RT-PCR法、Western bloting法检测细胞SDF-1 mRNA及蛋白,质粒转染24、48、72、96 h时MTT法测算三组细胞增殖能力(以光密度OD值表示),质粒转染48 h时观察三组细胞对奥沙利铂的耐药性(以奥沙利铂对细胞的半数抑制浓度IC 50 表示),质粒转染48 h时流式细胞仪测算三组细胞凋亡率,质粒转染48 h时Western blotting法检测三组细胞上皮型钙黏蛋白( E-cadherin)、神经型钙黏蛋白(N-cadherin)及波形蛋白(Vimentin)、微管相关蛋白1轻链3Ⅱ(LC3-Ⅱ)自噬相关因子(Beclin 1)。结果 与空白对照组、阴性对照组比较,培养48 h时实验组细胞SDF-1 mRNA及蛋白相对表达量均升高( P 均<0.05)。与空白对照组、阴性对照组比较,培养24、48、72、96 h实验组细胞OD值均升高( P 均<0.05)。质粒转染48 h时奥沙利铂对实验组、空白对照组、阴性对照组细胞的IC 50 值分别为 28.039 ±3.011、10.403±1.253、10.612±1.044(μg/mL),与空白对照组、阴性对照组比较,奥沙利铂对实验组细胞的IC 50 值升高( P 均<0.05)。质粒转染48 h时实验组、空白对照组、阴性对照组细胞凋亡率分别为39.10%± 2.26%、53.90%±3.19%、51.30%±3.37%,与空白对照组、阴性对照组比较,实验组细胞凋亡率降低( P 均< 0.05 )。质粒转染48 h时倒置显微镜下可见实验组细胞存在上皮-间质转化(EMT)现象,加入奥沙利铂后实验组细胞自噬活动增加;与空白对照组、阴性对照组比较,实验组细胞E-cadhein蛋白相对表达量降低,N-cadhein、Vimentin、LC3-Ⅱ、Beclin 1蛋白相对表达量均升高( P 均<0.05)。结论 SDF-Objective To observe the effects of stroma cell-derived factor-1 (SDF-1) gene transfection on the proliferation and drug resistance of human gastric cancer cell line SGC7901 and to explore its possible mechanism. Methods SGC7901 cells in the logarithmic growth phase were divided into the experimental group, negative control group, and blank control group. The cells in the experimental group and negative control group were transfected with pcDNA3.1-SDF-1 plasmid and pcDNA3.1 plasmid (blank plasmid), respectively, and the blank control group did not receive any treatment. After 48 h of plasmid transfection, SDF-1 mRNA and protein levels were detected by RT-PCR and Western bloting, respectively. The proliferation ability of the three groups was measured by MTT at 24, 48, 72 and 96 h after plasmid transfection (expressed by optical density). The drug resistance of the three groups to oxaliplatin was observed at 48 h after plasmid transfection (expressed by the 50% inhibitory concentration IC 50 of oxaliplatin). The apoptosis rates of the three groups were measured by flow cytometry at 48 h after plasmid transfection. After 48-hour plasmid transfection, Western blotting was used to detect epithelial cadherin (E-cadherin), neurocadherin (N-cadherin) and vimentin (Vimentin), microtubule-associated protein 1 light chain 3 Ⅱ(LC3-Ⅱ), autophagy-related factor (Beclin 1) in the three groups. Results The relative mRNA and protein expression levels of SDF-1 in the experimental group were significantly higher than those in the blank control group and negative control group at 48 h (both P <0 05). Compared with the blank control group and the negative control group, the OD values of the experimental group increased at 24, 48, 72 and 96 h (all P <0 05). At 48 h after plasmid transfection, the IC 50 values of oxaliplatin in the experimental group, blank control group and negative control group were (28.039±3.011),(10.403±1.253) and (10.612±1.044)μ g/mL, respectively. The experimental group had higher IC 50 value of oxalipl

关 键 词：基质细胞衍生因子-1 胃癌 胃癌细胞株SGC7901 细胞增殖 细胞耐药 上皮-间质转化 细胞自噬 

分 类 号：R735.2[医药卫生—肿瘤]