肝内及肝外胆管癌组织表达上调microRNAs表达谱的鉴定  

Identification of up-regulated miRNAs in extrahepatic and intrahepatic cholangiocarcinoma

作　　者：刘长征[1] 刘卫[2] 李静静[2] 郑毅[2] 于岚[2] 何小东[2] 陈松森[1] 

机构地区：[1]中国医学科学院基础医学研究所医学分子生物学国家重点实验室,北京100005 [2]中国医学科学院北京协和医院基本外科

出　　处：《中华肝胆外科杂志》2012年第6期466-469,共4页Chinese Journal of Hepatobiliary Surgery

基　　金：本课题受国家自然科学基金青年基金资助（基金编号81100608）

摘　　要：目的筛选、鉴定肝内及肝外胆管癌组织表达上调的miRNAs，并探讨其在胆管癌细胞增殖中的作用。方法利用miRNA-基因芯片方法筛选肝内、肝外胆管癌组织特异表达上调或共同表达上调的miRNAs；利用实时PCR方法对其进行验证；选择特异miRNA抑制剂转染胆管癌细胞系QBC939并利用MTT检测方法，对上述表达上调明显的miRNAs进行初步的功能研究，探讨miRNAs表达上调与胆管细胞癌细胞增殖的关系。结果12个miRNAs在肝外及肝内胆管癌组织共同表达上调，肝外胆管癌组织特异表达上调的miRNAs有28个，其中miR-125b与miR-19a分别表达上调3．7倍与3．6倍（P〈0．05）；肝内胆管癌有12个miRNAs特异表达上调，其中miR-92a与miR-205分别表达上调约4．5倍与3．5倍（P〈0．05）；在胆管癌细胞系QBC939中抑制miR-125b、miR-19a、miR-21以及miR-378’的内源性表达，可以明显抑制QBC939细胞的增殖，其抑制效率分别为71％、72％、69％与76％（P〈0．05，36h），61％、63％、60％与59％（P〈0．01，48h），61％、56％、60％与59％（P〈0．05，60h）。结论肝内及肝外胆管癌具有不同的miRNAs上调表达谱，敲低表达上调的miRNAs可以明显抑制胆管癌细胞增殖。Objective To investigate the expression profile of miRNAs up-regualted in human extrahepatic and intrahepatic cholangiocarcinoma tissues and probe the effect on cell growth of four of these miRNAs in QBC939 cell line. Methods Up-regulated miRNAs in extrahepatic or intrahepatic cholangiocarcinoma tissues were analyzed by using miRNA-microarray, which was confirmed by using miRNA Real-Time PCR analysis. Based on these findings, four of these up-regulated miNRAs were chosen to perform function investigation. The specific miRNA inhibitors were transfected into QBC939 cells, respectively, and cell proliferation assay was performed by using MTT. Results 12 miRNAs were up-regulated both in two types of cholangioeareinoma tissues, 28 miRNAs and 21 miRNAs were up-regulated in extrahepatie eholangioeareinoma and intrahepatic cholangioearcinoma, respectively. MiR-125b and miR-19a expression levels were increased about 3.7- and 3.6-fold, compared with the matched normal bile duct tissues （P〈0.05）. MiR-92a and miR-205 expression was upregulated about 4.5- and 3.5-fold, compared with the matched normal bile duct tissues （P〈0.05）. MiR-125b, miR- 19a, miR-21, and miR-378＊ were inhibited in QBC939 cells, which indicated a significant inhibitory effect on cell growth. The ratio of inhibition was 71%, 72%, 69%, and 76% （P〈0.05）at a6 h, 61%, 63%, 60%, and 59M（P〈0. 01） at 48 h, and 61%,56%,60% and 59z%（P〈0.05） at 60 h. Conclusion The miRNAs expression patterns in human extrahepatic and intrahepatic cholangiocarci- noma tissues are different and up-regulated miRNAs act as oncomirs on cholangiocarcinoma cell growth.

关 键 词：肝外胆管癌 肝内胆管癌 microRNA 细胞增殖 

分 类 号：R735.8[医药卫生—肿瘤]