摘要
目的 探讨过氧化物酶体增殖因子活化受体γ(Peroxisome Proliferator-actived Recepter PPARγ)配体罗格列酮(Rosiglitazone,RSG)体外抑制人肝癌SMMC7721细胞系细胞生长作用及初步探讨其作用机制。
方法 体外培养人肝癌SMMC7721细胞,以全反式维甲酸(All-trans retinoic,ATRA)为阳性对照药,MTT比色试验测定药物对细胞生长的影响,流式细胞术(Flow cytometry,FCM)检测细胞周期和细胞凋亡,分光光度法测定γ-GT变化,ELISA检测AFP分泌量,免疫细胞化学检测药物处理前后PPARγ,CyclinD_1蛋白表达的改变。
结果 不同浓度的RSG(12.5、25、50、100μmol/L)和10μmol/L的ATRA处理细胞后抑制肝癌SMMC7721细胞生长,RSG抑制肝癌SMMC7721细胞生长呈时间依赖性和浓度依赖性;γ-GT量明显下降,AFP分泌显著降低;G1期细胞数目增加,S期和G_2数目减少,而凋亡率无明显改变;细胞的PPARγ蛋白核内表达上调,CyclinD_1蛋白表达下调。
结论 过氧化物酶体增殖因子活化受体γ(PPARγ)配体罗格列酮(Rosiglitazone,RSG)具有抑制体外培养肝癌SMMC7721细胞生长作用;罗格列酮对SMMC7721细胞生长抑制作用与其阻滞细胞周期和诱导细胞分化相关;PPARγ蛋白核内表达上调、CyclinD_1蛋白表达下调可能介导了RSG抑制细胞生长。
Objective To investigate the effect of Rosiglitazone , the ligand of
Peroxisome Proliferator-activated Receptor gamma, on cell proliferation inhibition of hepatocarcinoma cells in vitro and its possible mechanism.
Methods Human hepatocarcinoma SMMC7721 cell line was cultured in
vitro, Cultured SMMC7721 cells were designed into 12.5 μ mol/L RSG group, 25 μ mol/L RSG group, 50 μ mol/L RSG group, 100 μ mol/L RSG group and the control group, 10 μ mol/L ATRA group. Proliferation inhibition effect was tested by MTT assay; Flow cytometry was used to detect cell cycle and apoptosis rate; The activity of γ -GT was measured with γ -GT kit and the secretive AFP was measured with ELISA ; Expression of PPARγ、 CyclinD_1 protein in hepatocarcinoma cells between untreated and treated with or without Rosiglitazone were observed by immunohistochemistry.
Results Rosiglitazone at various concentrations of 12.5、 25、 50、 100
μmol/L and All-trans retinoic acid at concentration 10 μmol·L~(-1) could inhibit the proliferation of SMMC7721 cells, RSG inhibit the proliferation in a dose-dependent and time-dependent manner; Cell micrographys tended to be normal after treatment; the specific activities of γ-GT and the secretive amount of AFP was reduced; Flow cytometry analysis reavealed a cell cycle arrest at G1 phase ,but apoptosis rate was not changed remarkably; nuclear traslocation and up-regulation of PPARγ protein expression and the down -regulation of CyclinD protein expression were observed .
Conclusion The ligand of PPARγ, Rosiglitazone induced significantly growth inhibition of human hepatocarcinoma SMMC7721 cells line in vitro which
引文
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