新维甲类化合物4HPR对人胆管癌细胞周期和凋亡的影响及可能机制

Induction of cell-cycle arrest and apoptosis by 4HPR in human cholangiocarcinoma cells and its possible mechanism

目的 探讨N hydroxyphenyl retinamide(4HPR)对人胆管癌细胞QBC939细胞增殖和凋亡的影响及其可能机制。方法 采用四甲基氮唑蓝实验 (MTT)、免疫组织化学染色、流式细胞术等技术 ,检测经 5× 10 -6mol/L 4HPR处理后 ,QBC939细胞生长增殖、细胞周期和凋亡的变化 ,以及对P2 1waf1、P2 7kip1、P5 3蛋白表达的影响。结果 4HPR显著抑制人胆管癌细胞QBC939细胞的生长增殖 ,并呈时间、剂量依赖性。用 5× 10 -6mol/L 4HPR处理细胞后 ,细胞周期停滞于G1期。流式细胞术检测发现 ,4HPR处理 12h细胞凋亡率达 9 7% ,至 4 8h达 5 7 2 %。而P2 1waf1、P2 7kip1蛋白表达呈上升趋势 ,P5 3蛋白表达则无明显变化。结论 4HPR具有抗增殖和诱导QBC939细胞发生凋亡的作用 ,并可导致细胞周期停滞于G1期。此种机制与其上调P2 1waf1、P2 7kip1蛋白表达有关 ,而与P5 3蛋白表达无关。

Objective To determine whether 4HPR can inhibit growth and induce apoptosis in human cholangiocarcinoma cell line QBC939. Methods MTT was used to evaluate the growth inhibiting effect of 4HPR on QBC939 cells. The expression of P53, P21 waf1 and P27 kip1 protein was determined with immunohistochemistry. The cell cycle and apoptosis in QBC939 cells were determined with flow cytometry. Results 4HPR significantly inhibited the proliferation of QBC939 cells in a time- and dose-dependent manner. Flow cytometric analysis indicated that treatment of QBC939 cells with 4HPR strongly induced cell cycle arrest at the G1 phase. The cell-cycle arrest was accompanied by an increase in expression of P21 waf1 and P27 kip1 protein but not P53. Meanwhile, the analysis showed that a marked reduction in the number of QBC939 cells with 4HPR was related to the induction of apoptosis. The percentage of apoptotic cells increased from 9.7% to 57.2% in 12 to 48 h after the treatment with 5×10 -6 M 4HPR. Conclusions 4HPR inhibits the growth of QBC939 cells by means of G_1 phase cell-cycle arrest resulting from up-regulation of P21 waf1 and P27 kip1 protein as well as the induction of apoptosis.