MIF和Cyclin D1在肝细胞癌中的表达  

The expression of MIF and Cyclin D1 in hepatocellular carcinoma

作　　者：夏金堂[1] 伍兆锋[1] 李雯[3] 赖越元[1] 赵杰[1] 徐晨[1] 王花[3] 滕元[3] 李瑜元[2] 

机构地区：[1]广州医学院附属广州市第一人民医院肝胆外科,510180 [2]广州医学院附属广州市第一人民医院消化内科,510180 [3]中山大学附属第一医院外科实验中心

出　　处：《中华普通外科杂志》2009年第5期398-401,共4页Chinese Journal of General Surgery

基　　金：广东省自然科学基金资助项目（5001776）,广州市科技计划基金资助项目（2005Z3-E0381）

摘　　要：目的探讨巨噬细胞游走抑制因子（macrophage migration inhibitory factor，MIF）和细胞周期蛋白D1（cyclinD1）在原发性肝细胞癌（hepatocellular carcinoma，HCC）组织中的表达及两者在肝癌发病机制及细胞周期调控中的关系。方法应用定量PCR及Western blot技术检测MIF和Cyclin D1在肝癌组织和癌旁组织的表达。化学合成MIF siRNA，将PLC细胞和Hep G2细胞分成对照组、MIF siRNA 50nmol/L干预组、MIF siRNA 100nmol/L干预组，脂质体法转染肝癌细胞PLC和HepG2，定量PCR技术和Western blot检测MIF siRNA干扰后MIF和Cyclin D1 mRNA和蛋白的表达变化。结果MIF和Cyclin D1蛋白在肝癌组织中的相对表达量为0．825±0．13和0．843±0．104：MIF和Cyclin D1 mRNA在肝癌组织中的相对表达量为癌旁组织的（7．31±1．85）倍和（4．27±1．05）倍，与癌旁组织相比，差异均具有统计学意义（FMIF=15．5，P〈0．01；Fcyclin D4=87．5，P〈0．01）。应用小RNA干扰技术转染PLC和HepG2肝癌细胞后MIF mRNA分别下调71．2％±7．2％、87．4％±2．9％、74．3％±8．9％、88．4％±4．6％（FPLC=315．5，P〈0．01；FHepG2=201．2，P〈0．01）；MIF蛋白分别下调为0．33±0．03、0．11±0．02、0．81±0．08、0．36±0．02，并呈剂量依赖关系（FPLC=43．9，P〈0．01；FHepG2=133．4，P〈0．01）。伴随MIF mRNA和蛋白的表达下调Cyclin D1 mRNA分别下调68．2％±3％、78．1％±1．4％、65．8％±4．7％、77．3％±2．6％（FPLC=1569，P〈0．01；FHepG2=480．4，P〈0．01）；Cyclin D1蛋白下调0．28±0．06、0．15±0．03、0．44±0．04、0．13±0．02，亦呈剂量依赖关系，与对照组相比差异有统计学意义（P〈0．01），两干预组相比差异有统计学意义（FPLC=35．5，P〈0．01；FHepG2=114．7，P〈0．01）。结论MIF和Cyclin D1在肝细胞癌中过表达，MIF可能在转录水平调控Cyclin D1的表达，并促使肿瘤细胞通过细胞周期检测点，�Objective To investigate the expression of macrophage migration inhibition factor （MIF） and cell cycle regulating factor Cyclin D1 in hepatoeellular carcinoma tissue and the interaction between MIF and Cyclin D1 in hepatocellular carcinoma cell cycle controlling. Methods Using quantitative real-time PCR and Western blotting to detect mRNA and protein expression of MIF and Cyclin D1 in HCC tissues and tumor adjacent tissues. Specific small interfering RNA（siRNA） targeting MIF gene was transfected at doses of 50 nmol/L and 100 nmoL/L into HCC cell lines of PLC and HepG2 with lipofectamine 2000 methods to knockdown the expression of MIF gene and to investigare the the interaction between MIF and Cyclin D1. Results MIF and Cyclin D1 protein and mRNA were overexpressed in HCC tumor tissues. The relative expression of MIF,Cyclin D1 protein and mRNA were 0. 825 ± 0. 13,0. 843 ± 0. 104 and 7.31 ± 1.85 folds,4. 27 ± 1.05 folds, compared with the tumor adjacent tissues （ FMEF= 15.5, P 〈0. 01 ; FCyclin Dl = 87.5, P 〈 0. 01 ）. In MIF siRNA treated PLC and HepG2 cells, MIF mRNA down regulation 71.2% ± 7. 2%, 87.4% ± 2. 9% , 74. 3% ± 8.9% and 88.4% ± 4. 6% respectively （ FpLc = 315. 5 ,P 〈 0.01 ; FHepG2 = 201.2 P 〈 0. 01 ）. While MIF protein expression were significantly reduced to 0. 33 ± 0.03,0. 11 ± 0. 02, 0. 81 ± 0. 08 and 0. 36 ± 0. 02 in a dose-dependent manner （ FPLC= 43.9, P 〈0. 01 ;FHepG2 = 133.4 P 〈 0. 01 ）. Cyclin D1 mRNA was significantly down-regulated in MIF siRNA treated PLC and HepG2 cell lines when compared with control group （ P 〈 0.01 ）. In 50 nmol/L and 100 nmol/L groups, Cyclin D1 mRNA levels were respectively decreased by 68. 2% ± 3% and 78. 1% ± 1.4% in PLC cell, 65.8% ±4.7% and 77.3% ±2.6% in HepG2 cell（FPLC=1569,P〈0.01;FHepG2 =480.4,P〈 0. 01 ）. Compared with control groups, Cyelin D1 protein levels significantly reduced to 0. 28 ± 0.06, 0. 15 ± 0. 03 and 0. 44 ± 0. 04,0. 13 _＋ 0. 02 in the PLC and HepG2 after MIF siRNA treatment（ FPLC

关 键 词：癌 肝细胞 巨噬细胞游走抑制因子 RNA干扰 细胞周期蛋白D1 

分 类 号：R686[医药卫生—骨科学]