stathmin蛋白促进人SMMC-7721肝癌细胞增殖侵袭能力  

Stathmin for promoting cell proliferation and invasion ability in human SMMC-7721 hepatocellular carcinoma cells

作　　者：龚舒[1] 陶忠桦[1] 刘晓燕[1] 郭坤[2] 刘银坤[2] 甘淋[1] 

机构地区：[1]泸州医学院医学基础研究中心,四川泸州646000 [2]复旦大学附属中山医院肝癌研究所,上海200032

出　　处：《重庆医学》2014年第10期1168-1170,1174,共4页Chongqing medicine

基　　金：四川省教育厅基金资助项目(11ZB125);泸州医学院自然科学基金资助项目(2012ZD-06;202;451)

摘　　要：目的构建微管解离蛋白stathmin过表达的人SMMC-7721肝癌细胞株,并探讨stathmin过表达对肝癌细胞增殖侵袭的影响。方法采用脂质体将Flag-pcDNA3.1-stathmin重组质粒和Flag-pcDNA3.1空质粒分别转染人SMMC-7221肝癌细胞,抗生素加压筛选,构建稳定表达stathmin的SMMC-7721细胞(实验组),以稳转空质粒的细胞为对照组,Western blot对建系细胞进行鉴定。采用CCK-8和软琼脂克隆形成实验检测细胞增殖;流式细胞术(FCM)检测细胞凋亡和细胞周期;体外细胞运动侵袭实验(Transwell)测定细胞的运动、侵袭能力。结果实验组中stathmin的表达0.76±0.12较对照组0.16±0.05明显增加(P<0.05),成功构建了过表达stathmin的人SMMC-7721肝癌细胞株;CCK-8和软琼脂克隆形成实验显示,实验组的细胞增殖较对照组明显增加(A4500.60±0.05 vs.0.29±0.03,P<0.01);实验组细胞凋亡降低[(5.80±0.33)%vs.(11.57±1.09)%,P<0.05],细胞周期阻滞在G2/M期;Transwell实验结果证实,与对照组比较,实验组细胞运动和侵袭能力均显著加强[运动实验透膜细胞数:(54.03±7.21)个vs.(130.45±14.13)个;侵袭实验透膜细胞数:(17.75±2.52)个vs.(57.76±8.50)个],差异均有统计学意义(P<0.01)。结论 stathmin的过表达能促进人SMMC-7721肝癌细胞的增殖侵袭能力。Objective To construct the stable stathmin-overexpression SMMC-7721 hepatocellular carcinoma cells and to explore the effect of stathmin-overexpression on the cell proliferation and metastasis in SMMC-7721 cells .Methods By using liposome , Flag-pcDNA3 .1 and Flag-pcDNA3 .1-stathmin plasmid were transfected into SMMC-7721 cells respectively ,the stable Flag-pcD-NA3 .1 expression cells（control group） and the stable stathmin-overexpression cells（experimental group） were established after an-tibiotic resistant gene screening ,and the cell lines were identified by Western Blot .Subsequently ,the cell proliferation was detected by cell count kit（CCK-8） and the soft agar assay ,the apoptosis and cell cycle were determined by the flow cytometry （FCM ） ,and the cell motility and invasion were analyzed by the Transwell assay in vitro .Results The stathmin protein expression of the experi-mental group was significantly increased compared with the control group （0 .76±012vs. 0 .12 vs .0 .16±0 .05 ,P〈0 .05） ,which indicated that the stathmin-overexpression human SMMC-7721 hepatocellular carcinoma cell line was successfully constructed .CCK-8 and the soft agar assay showed that the cell proliferation of the experimental group was higher than that of the control group （0 .29±0 .03 vs .0 .60±0 .05 ,P〈0 .01）;additionally ,the apoptotic ratio of the experimental group was decreased compared with the control group[（11 .57±1 .09）% vs .（5 .80±0 .33）% ,P〈0 .05] ,the cell cycle was arrested in the stage G2/M ;the Transwell experiment results verified that the cell motility and the invasive ability of the experimental group were obviously reinforced compared with the control group[transmenbrane cell numbers in migrant assay ：（54 .03±7 .21） vs .（130 .45±14 .13）;transmenbrane cell numbers in invasive assay ：（17 .75±2 .52） vs .（57 .76±8 .50） respectively] ,the differences had statistical significance（P〈0 .01） .Conclusion The overexpression of stathmin promotes the c

关 键 词：STATHMIN 肝肿瘤 细胞增殖 肿瘤浸润 肿瘤转移 

分 类 号：R735.7[医药卫生—肿瘤]