Effect Of Trichostatin A Combined With Rosiglitazone On The Proliferation And Invasion Inhibition Of Human Stomach Cancer Cells

Objective To investigate the effect of histone deacetylase (HDAC)inhibitor Trichostatin A combined with Peroxisome proliferator-activated receptorγ(PPARγ)agonist Rosiglitazone on the proliferation of human stomach cancer cells SGC-7901 as well as the mechanisms.Mechods SGC-7901 cells were routinely cultured in RPMI1640 medium supplemented with 10% heat-inactived fetal serum . Firstly,treat cells with different concentrations of TSA and ROZ respectively to select the best combination through assessing the growth of cells by MTT assay.Cells were randomly divided into control group,TSA group,ROZ group,combination group.The inhibitive rate were detected by MTT after 48 hours;Flow cytometric analysis was used to detected the cycle status of SGC-7901 cells. The expression level of PPARγand Cyclin D1 was assessed by reverse transcription-polymerase chain reaction(RT-PCR).Results MTT assay showed that low concentration of TSA could markedly sensitized SAGC-7901 cells to ROZ treatment compared with those treated ROZ only. The proportion of cells in G0/G1 phase treated with TSA and ROZ was higher than that with TSA and ROZ singly.The combination of TSA and ROZ remarkably reduced the expression of Cyclin D1 mRNA. We found that the expression of PPARγmRNA increased significantly after TSA treatment .Conclusion HDAC plays a negative role in activation of PPARγ. Trichostatin A combined with Rosiglitazone shows a synergic effect on the proliferation inhibition of human stomach cancer cells. Objective Histone deacetylase (HDAC) has previously been shown to attenuating peroxisome proliferators-activated receptor gamma's capacity to drive adipocyte differentiation. PPARγactivation is confirmed to inhibit the development and metastasis of a variety of malignant cell.This study was to investigate the role of HDAC in inhibiting the invasion of human gastric carcinoma SGC-7901 cells through PPARγmediated 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 the growth of cells by MTT assay. Then cells were randomly divided into control group, TSA group, ROZ group, combination group. The inhibition ratio was detected by MTT after 48 hours; Boyden chamber invasion test was used to detected the invasion ability of SGC-7901 cells in vitro. The mRNA level of PPARγand MMP2 was assessed by reverse transcription-polymerase chain reaction (RT-PCR), and the protein level of MMP2 was evaluated by Western blot.Results MTT assay showed that both TSA and ROZ inhibited SGC-7901 cells proliferation in a dose-dependent manner. In combined treatment with 20nmol/L TSA and 5μmol/L ROZ, a synergistic effect on inhibiting invasion of SGC-7901 cells was found. ROZ could down-regulate the mRNA and protein expression of MMP2, and the combination of TSA and ROZ reduced the expression of MMP2 mRNA and protein more than single ROZ .RT-PCR showed that TSA could up-regulate the mRNA expression of PPARγ.Conclusion HDAC plays a negative role in inhibiting the invasion of human gastric carcinoma cell lines by PPARγ-mediated pathway through limiting activation of PPARγ, and the invasion inhibition to human gastric carcinoma cell lines by ROZ will be enganced after the activity of HDAC is inhibited by TSA.