人肝门部胆管癌细胞QBC939中Vimentin基因的siRNA构建研究  

SiRNA construction of Vimentin gene in human hilar cholangiocarcinoma cell QBC939

作　　者：王磊 贾剑超 宋双龙 戚晓辉 胡向赟 王石 WANG Lei;JIA Jian-chao;SONG Shuang-long(Graduate School of Inner Mongolia Medical University,Huhhot 010110,China)

机构地区：[1]内蒙古医科大学研究生学院,内蒙古呼和浩特010110 [2]内蒙古自治区人民医院临床医学研究中心,内蒙古呼和浩特010110 [3]内蒙古自治区人民医院肝胆胰脾外科,内蒙古呼和浩特010110

出　　处：《中国实验诊断学》2022年第3期409-412,共4页Chinese Journal of Laboratory Diagnosis

基　　金：内蒙古自治区自然科学基金(2019MS08081);内蒙古科技计划项目(201602097)。

摘　　要：目的 构建可抑制Vimentin基因表达的siRNA并转染进入人肝门部胆管癌QBC939细胞株,初步鉴定siRNA的干扰效果。方法 设计并合成Vimentin(NM-003380)基因的可作用于不同位点的3种siRNA及一组siRNA阴性对照序列,使用脂质体法将3种不同的siRNA序列及阴性对照序列瞬时转染进入人肝门部胆管癌QBC939细胞,分别命名为T1组(siVIM-1)、T2组(siVIM-2)、T3组(siVIM-3)及NC组(siCtrl),使用RT-PCR、Q-PCR技术测定实验中各组细胞Vimentin在mRNA水平上的表达情况。结果 T1、T2及T3组Vimentin在基因表达水平上均较NC组受到抑制,T2组的抑制效果较T1及T3组更明显,差异具有统计学意义(P<0.01)。结论 成功建立并筛选出人肝门部胆管癌细胞QBC939细胞Vimentin基因的siRNA。为后续进一步研究Vimentin的生物学特性及在肝门部胆管癌QBC939细胞中的作用奠定了基础。Objective SiRNA that could inhibit Vimentin gene expression was constructed and transfected into human hilar cholangiocarcinoma QBC939cell line to preliminarily identify the interference effect of siRNA.Methods Three kinds of siRNA of Vimentin(NM-003380)gene that can act on different sites and a set of siRNA negative control sequence were designed and synthesized.Three kinds of siRNA sequence and negative control sequence were transfected into human hilar cholangiocarcinoma QBC939cells by liposome method.They were named T1group(siVIM-1),T2group(siVIM-2),T3group(siVIM-3)and NC group(siCtrl).The expression of Vimentin at mRNA level in each group was determined by RT-PCR and Q-PCR techniques.Results The gene expression levels of Vimentin in T1,T2and T3groups were inhibited compared with those in NC group,and the inhibitory effect of T2group was more obvious than that of T1and T3groups,the difference was statistically significant(P<0.01).Conclusion SiRNA of Vimentin gene in human hilar cholangiocarcinoma QBC939cells was successfully established and screened.This laid a foundation for further study of the biological characteristics of Vimentin and its role in hilar cholangiocarcinoma QBC939cells.

关 键 词：肝门部胆管癌 QBC939细胞 SIRNA 

分 类 号：R735.8[医药卫生—肿瘤]