反义CD147分子对人胆管癌细胞株侵袭性影响的实验研究  

Effect of antisense Cd147 on invasion of human cholangiocarcinoma cell line QBC939

作　　者：易为民[1] 彭创[1] 厉鸥[1] 郭超[1] 黄喆[1] 谭朝霞[1] YI Wei-min;PENG Chuang;LI Ou;GUO Chao;HUANG Zhe;TAN Chao-xia(the Eighth Ward of Liver and Gallbladder, Hunan Provincial People’s Hospital, Changsha 410005, Hunan)

机构地区：[1]湖南省人民医院肝胆8病室,湖南长沙410005

出　　处：《肝胆胰外科杂志》2017年第2期138-144,共7页Journal of Hepatopancreatobiliary Surgery

基　　金：湖南省科学技术厅科技计划一般项目(2012FJ4105)

摘　　要：目的探讨反义CD147分子对人胆管癌细胞株QBC939侵袭性的影响。方法构建含反义CD147分子片段的重组真核表达质粒,将人胆管癌细胞株QBC939分为三组:(1)反义CD147组,运用重组真核表达质粒as CD147-pc DNA3.1(-)转染QBC939细胞;(2)空质粒组,运用pc DNA3.1(-)空质粒转染QBC939细胞;(3)对照组,运用DMEM常规处理细胞。采用MTT法检测各组QBC939细胞的生长情况,RT-PCR和Western blotting法分别对三组QBC939细胞中的CD147、MMP-2、MMP-9、TIMP-2的m RNA表达水平及其对应蛋白的表达水平进行检测。结果成功构建了携带反义CD147分子片段的重组真核表达质粒。MTT法检测显示:三组QBC939细胞的生长曲线及对数生长期情况无统计学差异(P>0.05)。RT-PCR法检测显示:反义CD147组QBC939细胞中CD147和MMP-2分子的m RNA表达均低于空质粒组和对照组(P<0.05),且空质粒组与对照组相比无统计学差异(P>0.05);反义CD147组QBC939细胞中MMP-9和TIMP-2分子的m RNA表达与空质粒组和对照组相比无统计学差异(P>0.05)。Western blotting检测发现:与空质粒组和对照组相比,反义CD147组QBC939细胞中CD147和MMP-2分子的蛋白表达明显降低(P<0.05),空质粒组与对照组相比无统计学差异(P>0.05);与空质粒组和对照组相比,反义CD147组QBC939细胞株中MMP-9和TIMP-2分子的蛋白表达无统计学差异(P>0.05)。结论 (1)反义CD147对胆管癌细胞株QBC939的生长无影响;(2)反义CD147对胆管癌细胞株QBC939中MMP-9和TIMP-2的m RNA表达及对应蛋白表达无影响;(3)反义CD147降低胆管癌细胞株QBC939中CD147和MMP-2的m RNA表达及对应蛋白表达,可能会降低胆管癌细胞株的侵袭性。Objective To investigate the effect of antisense CD147on invasion of human cholangiocarcinomacell line(QBC939).Methods A eukaryotic expression vector containing the CD147cDNA in an antisenseorientation was reconstructured.Human cholangiocarcinoma cell were divided into either a antisense CD147treated group,empty vector group,or control group.QBC939in antisense CD147treated group was undergonetransfection with CD147-pcDNA3.1(-);QBC939in empty vector group was undergone transfection with pcDNA3.1(-);and control group was treated with DMEM.Three groups were tested with MTT method for cell proliferation.Real-time PCR was used to detect the mRNA expression of CD147,MMP-2,MMP-9and TIMP-2.Westernblotting was used to detect the protein expression of CD147,MMP-2,MMP-9and TIMP-2.Results A eukaryoticexpression vector containing the CD147cDNA in an antisense orientation was successfully reconstructed.In treated group,no reduction in the growth rate was found after infection with antisense CD147(P>0.05).Compared with empty vector group and control group,the mRNA expression of CD147and MMP-2moleculewas effectively inhibited in the antisense CD147treated group(P<0.05),and there was no significant differencebetween empty vector group and control group(P>0.05).For mRNA expression of MMP-9and TIMP-2molecule,there was no significant difference between the antisense CD147treated group and empty vector group andcontrol group(P>0.05).Compared with empty vector group and control group,the protein expression of CD147and MMP-2molecule was effectively inhibited in the antisense CD147treated group(P<0.05),and there was nosignificant difference between empty vector group and control group(P>0.05).For protein expression of MMP-9and TIMP-2molecule,there was no significant difference between the antisense CD147treated group and emptyvector group and control group(P>0.05).Conclusion(1)Antisense CD147has no effect on proliferation ofQBC939cell.(2)Antisense CD147has no effect on the mRNA expression and protein expression of MMP-9andTIMP-2molecule

关 键 词：反义CD147 基质金属蛋白酶 基质金属蛋白酶抑制剂2 人胆管癌细胞株QBC939 

分 类 号：R735.8[医药卫生—肿瘤]