Ma Guo-Dong, Zhang Cai-Quan, Hou Jun, Zhang Long-Yun, Lin Chao
Department of General Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing, People's Republic of China.
Ai Zheng. 2009 Jul;28(7):695-701. doi: 10.5732/cjc.009.10014.
Histone deacetylase (HDAC) can attenuate the function of peroxisome proliferator-activated receptor gamma (PPARgamma) to drive adipocyte differentiation. PPARgamma activation is confirmed to inhibit the development and metastasis of a variety of malignant cells. This study was to investigate the role of HDAC in inhibiting the invasion of human gastric carcinoma SGC-7901 cells through PPARgamma-mediated pathway, and explore potential mechanism.
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 PPARgamma and matrix metalloproteinase-2 (MMP-2) were assessed by reverse transcription-polymerase chain reaction (RT-PCR), and the protein level of MMP-2 was evaluated by Western blot.
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 mumol/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 PPARgamma mRNA.
HDAC suppresses the activation of PPARgamma 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.
组蛋白去乙酰化酶（HDAC）可减弱过氧化物酶体增殖物激活受体γ（PPARγ）驱动脂肪细胞分化的功能。PPARγ激活已被证实可抑制多种恶性细胞的发展和转移。本研究旨在探讨HDAC通过PPARγ介导的途径在抑制人胃癌SGC - 7901细胞侵袭中的作用，并探索潜在机制。
分别用不同浓度的曲古抑菌素A（TSA）和罗格列酮（ROZ）处理SGC - 7901细胞，通过MTT法评估细胞增殖以选择最佳组合。然后将细胞随机分为对照组、TSA组、ROZ组和联合组。48小时后通过MTT法检测细胞增殖；通过Boyden小室侵袭试验检测细胞侵袭。通过逆转录 - 聚合酶链反应（RT - PCR）评估PPARγ和基质金属蛋白酶 - 2（MMP - 2）的mRNA水平，通过蛋白质印迹法评估MMP - 2的蛋白质水平。
TSA和ROZ均以剂量依赖性方式抑制SGC - 7901细胞的增殖。20 nmol/L TSA与5 μmol/L ROZ联合使用可协同抑制SGC - 7901细胞的侵袭（q = 1.41）。ROZ下调MMP - 2的mRNA和蛋白质表达。TSA与ROZ联合使用比单独使用ROZ更明显地降低MMP - 2表达。TSA上调PPARγ mRNA的表达。
HDAC通过一系列分子机制抑制PPARγ的激活。TSA抑制HDAC活性后，ROZ抑制人胃癌细胞侵袭的活性可增强。
