肝细胞肝癌SMMC-7721细胞中FOXQ1基因沉默对奥沙利铂的化疗增敏作用  

Enhanced sensitivity of hepatocellular carcinoma cell line SMMC-7721 to oxaliplatin by silencing forkhead box Q1 gene with RNA interference technology

作　　者：邓大炜 吴斌 严舒[1,2] 张光年 兰川[1,2] 弋鹏圣 曾丽娟[3] 李建水 

机构地区：[1]川北医学院肝胆胰肠研究所,四川南充637000 [2]川北医学院附属医院肝胆外科,四川南充637000 [3]川北医学院附属医院信息科,四川南充637000

出　　处：《中国普外基础与临床杂志》2018年第3期289-295,共7页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的观察沉默肝癌SMMC-7721细胞中叉头框Q1(FOXQ1)基因的表达对奥沙利铂敏感性的影响。方法 (1)构建靶向FOXQ1基因的shRNA重组慢病毒载体及阴性对照重组慢病毒载体,再筛选最佳的干扰序列。(2)将细胞分为3组:干扰组(转染FOXQ1-sh RNA-1重组慢病毒载体)、阴性对照组(转染阴性对照重组慢病毒载体)及空白对照组(未做任何处理)。检测3组细胞中FOXQ1 mRNA及其蛋白的表达。(3)将另一部分细胞分为干扰组、阴性对照组、空白对照组、干扰+奥沙利铂、阴性对照+奥沙利铂组与空白对照+奥沙利铂组,给予相应处理,培养48 h后检测细胞凋亡率。细胞活力实验方法同细胞凋亡率实验。结果 (1)FOXQ1-shRNA-1组细胞中FOXQ1 mRNA及其蛋白的表达水平均低于FOXQ1-shRNA-2组和FOXQ1-shRNA-3组(P<0.05),为最佳干扰序列。(2)干扰组细胞中FOXQ1 mRNA及其蛋白的表达水平均低于阴性对照组和空白对照组(P<0.05),但阴性对照组和空白对照组比较差异均无统计学意义(P>0.05)。(3)不管是在加入OXA条件下,还是在未加入OXA条件下,干扰组细胞的凋亡率均高于阴性对照组与空白对照组(P<0.05),细胞活力均低于阴性对照组和空白对照组(P<0.05),但同条件下阴性对照组和空白对照组的细胞凋亡率和细胞活力比较差异均无统计学意义(P>0.05);在干扰组、阴性对照组和空白对照组中,均是加入OXA组的细胞凋亡率高于未加入OXA组(P<0.05),加入OXA组的细胞活力低于未加入OXA组(P<0.05)。结论沉默FOXQ1基因的表达能够有效诱导SMMC-7721细胞的凋亡,并增加其对奥沙利铂的化疗敏感性。Objective To observe the effect offorkhead box QI（FOXQ1） short hairpin RNA （shRNA） on sensitivity of oxaliplatin chemotherapy in hepatpcellular carcinoma cell line SMMC-7721. Methods ① Complementary shRNA oligonucleotides targeting the FOXQ 1 gene and negative control-shRNA were designed and inserted into lentiviral vector, shRNA lentivirus vectors were transfected into SMMC-7721 cells and the lentivirus vector with the best silencing effect was screened. ② SMMC-7721 cells were divided into interference group （SMMC-7721 cells were transfected with FOXQ 1-shRNA-1）, negative control group （SMMC-7721 cells were transfected with negative control-shRNA）, and blank control group （SMMC-7721 cells did not received any treatment）, and the expressions of FOXQ1 mRNA and its protein in SMMC-7721 cells were detected at 72 hours after transfection. ③ SMMC-7721 cells were divided into interference group, negative control group, blank control group, interference＋oxaliplatin group, negative control＋oxaliplatin group,and blank control＋oxaliplatin group, apoptosis rates and viability of SMMC-7721 cells were detected at 48 hours after transfection. Results ① The expressions ofFOXQ1 mRNA and its protein in SMMC-7721 cells of the FOXQI-shRNA-1 group were both lower than those of the FOXQI-shRNA-2 group and FOXQI-shRNA-3 group （P〈0.05）, so the FOXQ1- shRNA-1 was the best lentiviral vector. （~） Compared with the negative control group and the blank control group, the expressions of FOXQ1 mRNA and its protein of the interference group were both lower （P〈0.05）, but there was no significant difference between the negative control group and the blank control group （P〉0.05）. ③ Whether added oxaliplatin or not, compared with the negative control group and the blank control group, the apoptosis rates of the interference group were higher （P〈0.05）, but the viability of the interference group was lower （P〈0.05）, and there was no significant difference between the negative cont

关 键 词：肝细胞肝癌 叉头框Q1 奥沙利铂 增敏作用 

分 类 号：R735.7[医药卫生—肿瘤]