茶多酚对缺氧诱导的胆管癌细胞QBC939增殖及凋亡的影响与机制

Effects and mechanism of epigallocatechin-gallate on proliferation and apoptosis of cholangiocarcinomas cell line QBC939 under hypoxia

目的建立缺氧模型,观察EGCG对缺氧诱导的QBC939胆管癌细胞增殖、凋亡与转移的影响及其与VEGF表达的关系。方法 QBC939低氧培养,MTT观察EGCG对缺氧诱导的胆管癌细胞增殖的影响,过河实验观察EGCG对缺氧诱导的胆管癌细胞迁移的影响,用流式细胞术观察缺氧对细胞周期和细胞凋亡率的影响,利用ELISA观察缺氧诱导对胆管癌细胞VEGF表达的影响。结果 EGCG呈浓度和时间依赖性抑制缺氧诱导的胆管癌增殖;缺氧胆管癌细胞过河时间为(15.2±0.94)h,较常氧组(23.1±2.14)h显著缩短,EGCG(40、80和160μmol/L)使缺氧胆管癌细胞过河时间显著延长([24.1±2.34)、(36.7±3.27)和(47.8±3.04)h];缺氧胆管癌细胞经EGCG(40、80和160μmol/L)处理后,G1期细胞和凋亡率显著增加,同时伴有S期和G2期细胞减少;缺氧诱导组VEGF表达量为(2 515.2±190.1)Pg/mL,EGCG(40、80和160μmol/L)处理组,VEGF合成显著下降([2 419.8±231.7)、(1 736.7±203.2)和(935.7±125.4)Pg/mL]。结论 EGCG可抑制胆管癌细胞的增殖与迁移,其机制与下调VEGF表达有关。

【Objective】 To establish the hypoxia model to observe the effect of EGCG on QBC939 cell proliferation and VEGF expressions induced by hypoxia.【Methods】 The human cholangiocarcinomas line QBC939 cells were cultured under hypoxic conditions in vitro and MTT test was used to observe the influence of EGCG to hypoxic QBC939 cell.The crossing-river test was used to examine the invasiveness of QBC939.Cell cycle and the apoptosis were analyzed by flow cytometry（FCM）.ELISA was performed to determine the VEGF expression of hypoxic QBC939 cell.【Results】 EGCG inhibited the proliferation of QBC939 cell in dependent dose and time way.The crossing-river time of hypoxic QBC939 was shortened significantly,and EGCG Treatment increased the ecrossing-river time.The percentage of cells in G1 phase and the apoptosis began to increase after an EGCG treatment,and S and G2 phase increased significantly.The VEGF expression decreased in EGCG groups.【Conclusion】 EGCG can effectively inhibit the proliferation of QBC939 cells.The underlying mechanism may be due to the down-regulation of VEGF,which lead to a cell cycle arrest in G0/G1 phases,and finally inhibit the proliferation.