HDAC在PPARγ途径抑制胃癌SGC-7901细胞侵袭中的作用  

Role of histone deacetylase in inhibiting invasion of human gastric carcinoma cell line SGC-7901 by PPARγ-mediated pathway

作　　者：马国栋[1] 张才全[1] 侯俊[1] 张龙云[2] 蔺超[3] 

机构地区：[1]重庆医科大学附属第一医院普外科,重庆400016 [2]重庆医科大学附属第一医院妇产科,重庆400016 [3]重庆医科大学附属第二医院骨科,重庆400000

出　　处：《癌症》2009年第7期695-701,共7页Chinese Journal of Cancer

摘　　要：背景与目的:有研究表明组蛋白去乙酰化酶(histone deacety lase,HDAC)能抑制过氧化物酶体增殖物激活受体-γ(peroxisome proliferator-activated receptorγ,PPARγ)促进脂肪细胞分化的能力;最近发现PPARγ活化后,能够抑制多种肿瘤细胞的生长与侵袭。本研究旨在观察HDAC在PPARγ途径影响胃癌细胞株SGC-7901侵袭能力中的作用及机制。方法:用不同浓度的HDAC抑制剂曲古抑菌素A(TSA)和PPARγ配体罗格列酮(ROZ)分别作用于SGC-7901细胞,用四甲基偶氮唑蓝(MTT)法检测两种药物对细胞增殖的抑制作用,以筛选出合适的联合浓度。将SGC-7901细胞随机分为对照组、TSA组、ROZ组和TSA与ROZ联合组。MTT法检测细胞增殖抑制率,Transwell小室法检测各组胃癌细胞侵袭能力,RT-PCR法检测各组细胞PPARγ、MMP-2mRNA表达情况,Westernblot检测MMP-2蛋白的表达。结果:5μmol/LROZ和40nmol/LTSA为无明显细胞毒性作用的最高浓度,低于此浓度的ROZ及TSA可减弱SGC-7901细胞的侵袭能力(P<0.01)。5μmol/LROZ与20nmol/LTSA联合作用对SGC-7901细胞的侵袭抑制呈协同作用(q=1.41>1.15)。ROZ可以下调SGC-7901细胞MMP-2mRNA和蛋白的表达(P<0.01),TSA与ROZ联合处理组细胞MMP-2表达下调较单用ROZ组明显(P<0.01)。RT-PCR结果显示,TSA作用SGC-7901细胞后,可使PPARγ表达增加(P<0.01)。结论:HDAC通过一系列分子机制限制PPARγ的激活,应用TSA抑制HDAC活性后,可以使ROZ抑制胃癌细胞的侵袭活性增强。Background and Objective： Histone deacetylase （HDAC） can attenuate the function of peroxisome proliferator-activated receptor gamma （PPARγ） to drive adipocyte differentiation. PPARγ activation is confirmed to inhibit the development and metastasis of a variety of malignant ceils. This study was to investigate the role of HDAC in inhibiting the invasion of human gastric carcinoma SGC-7901 cells through PPARγ-mediated pathway, and explore potential mechanism. Methods： SGC-7901 cells were treated with different concentrations of Trichostatin A （TSA） and Rosiglitazone （ROZ） respectively to select the best combination through assessing cell proliferation by MTT assay. Then cells were randomly divided into control group, TSA group, ROZ group, and combination group. Cell proliferation was detected by MTT assay after 48 h; cell invasion was detected by Boyden chamber invasion test. The mRNA levels of PPARγ and matrix metalloproteinase-2 （MMP-2） were assessed by reverse transcriptionpolymerase chain reaction （RT-PCR）, and the protein level of MMP-2 was evaluated by Western blot. Results： Both TSA and ROZ inhibited the proliferation of SGC-7901 cells in a dose-dependent manner. A combination of 20 nmol/L TSA and 5 μmol/L ROZ synergistically inhibited the invasion of SGC-7901 cells （q=1.41）. ROZ down-regulated the mRNA and protein expression of MMP-2. TSA and ROZ in combination reduced MMP-2 expression more obviously than ROZ alone. TSA up-regulated the expression of PPARγ mRNA. Conclusions： HDAC suppresses the activation of PPARγ through a series of molecular mechanisms. The activity of ROZ in inhibiting invasion of human gastric carcinoma cells can be enhanced after the activity of HDAC is inhibited by TSA.

关 键 词：胃肿瘤 SGC-7901细胞 组蛋白去乙酰化酶 过氧化物酶体增殖物激活受体Γ 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]