论文部分内容阅读
目的探讨过氧化物酶增殖物活化受体γ在肝肿瘤细胞株中的表达,结合配体对其生长的影响,并初步探讨其机制。方法RT-PCR及Westen blot检测肝肿瘤细胞株中PPARγ的表达;MTT法检测PPARγ活化对肝肿瘤细胞生长的影响,Annexin V-FITC、DAPI染色、流式细胞仪、RT- PCR分别研究PPARγ对肝肿瘤细胞凋亡、细胞周期、周期调节因子影响。结果人类肝肿瘤细胞株SMMC-7721、Hep G2、Hep 3B在mRNA及蛋白水平均表达PPARγ;PPARγ/活化配体曲格列酮、吡格列酮作用肝肿瘤细胞株48 h后在25、50μmol/L浓度时显示明显的生长抑制作用,流式细胞仪显示细胞周期中G_0/G_1期显著延长,S期明显减少,曲格列酮10、25、50μmol/L作用SMMC-7721 48 h后,Cyclin D1的表达明显下调,P21~(WAF1/CIP1)的表达则明显上调,P27~(KIP1)未见明显变化。PPARγ配体在25、50μmol/L浓度时可诱导SMMC一7721凋亡。结论PPARγ在SMMC-7721、Hep G2、Hep 3B中存在表达;PPARγ配体对于肝肿瘤细胞株有明显的生长抑制作用,其生长抑制作用可能是通过下调Cyclin D1,上调P21~(WAF1/CIP1)表达,产生细胞周期阻滞及促进细胞凋亡而实现。
Objective To investigate the expression of peroxisome proliferator - activated receptor γ in liver tumor cell lines, and to study the effect of ligand on its growth and its mechanism. The expression of PPARγ in liver tumor cell lines was detected by RT-PCR and Westen blot. The effect of PPARγ activation on the growth of hepatic tumor cells was detected by MTT assay. Annexin V-FITC, DAPI staining, flow cytometry and RT- Liver Tumor Cell Apoptosis, Cell Cycle, Cycle Regulator. Results The human hepatocellular carcinoma cell lines SMMC-7721, Hep G2 and Hep 3B both expressed PPARγ at mRNA and protein levels. After treatment with PPARγ / activated ligand troglitazone and pioglitazone for 48 h, the cells were cultured at 25 and 50 μmol / L G_0 / G_1 phase was significantly prolonged in cell cycle and S phase was significantly decreased by flow cytometry. After 48 h SMMC-7721 treatment with troglitazone, 50 and 50 μmol / L, the expression of Cyclin D1 The expression of P21 (WAF1 / CIP1) was significantly up-regulated, but there was no obvious change in P27 (KIP1). PPARγ ligand can induce apoptosis of SMMC-7721 cells at the concentration of 25 and 50 μmol / L. Conclusions PPARγ is expressed in SMMC-7721, Hep G2 and Hep 3B. PPARγ ligand has obvious growth inhibitory effect on hepatocellular carcinoma cell line. Its growth inhibition may be through down-regulation of Cyclin D1, up-regulation of P21 WAF1 / CIP1, Expression, produce cell cycle arrest and promote apoptosis.