KiSS-1与肝癌细胞增殖及黏附和侵袭关系的体外实验研究  

An in vitro study of the relationship between KISS-1 expression and hepatoma carcinoma cell proliferation, adhesion, and invasion

作　　者：徐玫芳[1] 臧盛兵[2] 刘景丰[3] 高凌云[2] 高美钦[2] 杨映红[1] 黄爱民[2] 

机构地区：[1]福建医科大学附属协和医院病理科,福州350004 [2]福建医科大学基础医学院病理学系肿瘤研究室 [3]福建医科大学附属第一医院肝病中心

出　　处：《中华肝脏病杂志》2012年第12期925-929,共5页Chinese Journal of Hepatology

基　　金：基金项目：福建省教育厅基金（JS06011）

摘　　要：目的观察KiSS-1基因表达对肝癌细胞体外增殖、黏附及侵袭能力的影响，为进一步探讨其抗肝细胞癌侵袭转移的机制奠定基础。方法培养具有高转移潜能的人肝癌细胞株MHCC97-H，瞬时转染KiSS-1基因的细胞为实验组，转染空载体pcDNA3．1／HiSC的细胞为空白对照组，未转染细胞为阴性对照组，采用流式细胞术与四甲基偶氮唑盐法、基质黏附实验、Transwell体外侵袭和趋化运动实验检测KISS-1表达对MHCC97-H细胞体外增殖、黏附、侵袭和运动能力的影响。应用SPSS13．0统计软件包进行统计分析，组间差异以两样本t检验分析。结果转染组、空载体组和未转染组与Matrigel基质的黏附能力（A值）分别为0．257±0．029、0．374±0．016和0．394±0．031，转染组明显低于空载体组（t=-7．90345，P〈0．01）和未转染组（t=-7．22752，P〈0．01）；与Fibronectin基质的黏附能力（A值）分别为0．292±0．004、0．394±0．010和0．412±0．023，转染组明显低于空载体组（t=-20．93138，P〈0．01）和未转染组（t=-11．31371，P〈0．01）;趋化运动至Transwell小室下室表面的细胞数分别为65．80±1．92、93．80±2．28和96．40±2．07，转染组明显低于空载体组（t=-30．11750，P〈0．01）和未转染组（t=-24．19142，P〈0．01）;侵袭至Transwell小室下室表面的细胞数为42．40±1．14、66．00±1．58和67．80±1．92，转染组明显低于空载体组（t=-27．0711，P〈0．01）和未转染组（t=一25．4，P〈0．01）。而转染组、空载体组和未转染组的细胞增殖能力（A值）分别为0．644±0．027、O．669±0．022和0．678±0．027，转染组略低于空载体组（t=一1．60371，P〉0．05）和未转染组（t=-1．97828，P〉0．05）}同时，转染组较空载体组和未转染组未出现明显的G1期阻滞和S期阻滞，也未出现明显的凋亡峰。结论KISS-1表达虽不影响肝癌细胞的体外增殖能�Objective To investigate the impact of expression of kisspeptin-1 （KiSS-1） metastasissuppressor gene on the proliferative, adhesive and invasive abilities of human hepatocellular carcinoma （HCC） using an in vitro cell system. Methods The highly metastatic human hepatoma cell line MHCC97-H was transiently transfected with the pcDNA3.1/HisC vector expressing the KISS-1 gene （experimental group） or the vector without the KisS-I gene （blank control group）. Untransfected ceils served as the negative control group. Proliferative abilities of the three groups were assessed by flow cytometry and MTT assay. Adhesive abilities were assessed by MTT assays using matfigel and fibronectin. Invasive abilities and cell motility were assessed by chemoinvasion chamber assay using reconstituted matrigel and migration chamber assay using polycarbonate filters, respectively. Results The experimental group showed significantly lower adhesion capacity to matt±gel （0.257 ± 0.029） than either the blank control group （0.374 ± 0.016; t = -7.90345, P 〈 0.01） or the negative control group （0.394 ± 0.031; t= -7.22752, P 〈 0.01）. Similarly, the experimental group showed significantly lower adhesion capacity to fibronectin （0.292 ± 0.004） than either the blank control group （0.394 ± 0.010; t = -20.93138, P〈 0.01） or the negative control group （0.412 ± 0.023; t= -11.31371, P〈 0.01）. The experimental group also showed significantly lower numbers of cells with invasive capacity （42.40 ± 1.14） than either the blank control group （66 ± 1.58; t = -27.0711,P 〈 0.01 ） or the negative control group （67.80 ± 1.92; t = -25.4,P 〈 0.01）. Similarly, the experimental group showed significantly lower numbers of cells with chemotactic movement （65.80 ± 1.92） than either the blank control group （93.80 ± 2.28; t = -30.11750,P〈 0.01） or the negative control group （96.40 ± 2.07; t = -24.19142, P 〈 0.01 ）. The experimental group showed slightly, but not significantly, lower

关 键 词：癌 肝细胞 细胞增殖 基因 肿瘤抑制 

分 类 号：R735.7[医药卫生—肿瘤]