曲古抑菌素A与罗格列酮联合应用对胃癌细胞增殖的抑制作用

Inhibitory Effect of Trichostatin A Combined with Rosiglitazone on Proliferation of Stomach Cancer Cells

目的研究组蛋白去乙酰化酶(HDAC)抑制剂曲古抑菌素A(TSA)与过氧化物酶体增殖物激活受体γ配体罗格列酮(ROZ)联合应用对胃癌细胞SGC-7901增殖的抑制作用,并探讨其可能的作用机制。方法首先用不同浓度的TSA和ROZ分别作用于SGC-7901细胞,MTT法检测二者对细胞增殖的抑制作用,以筛选出合适的联合作用浓度。将SGC-7901细胞随机分为对照组、TSA组、ROZ组和TSA+ROZ组,加入相应的药物处理48h后,MTT法检测各组细胞增殖抑制率,并应用流式细胞仪检测细胞周期情况,RT-PCR检测PPARγ和CyclinD1基因mRNA的转录水平。结果对SGC-7901细胞增殖抑制作用不显著的低剂量TSA(40nmol/L)与ROZ联用时,能显著增强ROZ对细胞增殖的抑制作用,TSA+ROZ组G0/G1期细胞比例较单用TSA或ROZ组明显增加,且CyclinD1基因mRNA转录水平明显下降;TSA作用于细胞后,PPARγ基因mRNA转录水平较对照组明显升高,而ROZ单独作用后,细胞PPARγ基因mRNA转录水平无改变。结论TSA可明显增强ROZ对SGC-7901细胞增殖的抑制效应,提示HDAC在SGC-7901细胞内可能具有抑制PPARγ的作用。

Objective To investigate the inhibitory effect of histone deacetylase（HDAC）inhibitor trichostatin A（TSA）combined with peroxisome proliferator-activated receptor γ （PPARγ）agonist rosiglitazone （ROZ）on the proliferation of human stomach cancer SGC-7901 cells as well as its potential mechanism. Methods SGC-7901 cells were treated with TSA and ROZ,at various concentrations,respectively. The inhibitory effect of TSA or ROZ administered alone on cell proliferation was determined by MTT method to optimize the concentrations of the two drugs for combined administration. SGC-7901 cells were divided into control,TSA,ROZ and TSA ＋ ROZ groups randomly. The cells in TSA,ROZ and TSA＋ROZ groups were treated with the corresponding drugs,while those in control group were untreated. Forty-eight hours after treatment,the inhibiting rate of cell proliferation was determined by MTT method,the cell cycle by flow cytometry,and the transcription levels of PPARγ and Cyclin D1 mRNAs by RT-PCR. Results The TSA at a concentration of 40 nmol / L showed no significantly inhibitory effect on the proliferation of SGC-7901 cells when admin-istered alone. However,the TSA at the same concentration combined with ROZ enhanced the inhibitory effect of ROZ on cell prolifer-ation significantly. Compared with those in TSA and ROZ groups,the percentage of SGC-7901 cells at G0 / G1 stages in TSA ＋ ROZ group increased significantly,while the transcription level of Cyclin D1 mRNA decreased significantly. The transcription level of PPARγ mRNA was significantly higher in TSA group than in control group,but showed no significant difference in ROZ and control groups. Conclusion TSA enhanced the inhibitory effect of ROZ on proliferation of SGC-7901 cells significantly,indicating potentially inhibitory effect of HDAC in SGC-7901 cells on PPARγ.