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目的:探讨绿茶提取物(green tea extract,GTE)对人卵巢癌SKOV3细胞株生长增殖的影响及其作用途径。方法:应用MTT法观察SKOV3细胞生长增殖的情况;光学显微镜观察SKOV3细胞的形态学变化;流式细胞术(FCM)检测SKOV3细胞周期及凋亡的情况;DAPI标记法观察细胞凋亡形态变化;Western blotting检测p-ERK1/2、ERK1/2、Caspase-3及Bcl-xl等相关蛋白的表达水平。结果:随GTE浓度的增加及作用时间的延长,SKOV3细胞存活率逐渐降低(P<0.01),SKOV3细胞数目逐渐减少,细胞受损逐渐增多;经GTE(80μg/L)处理24h、48h、72h的SKOV3,G0/G1期细胞比例与对照组比均显著减少(P<0.01),S期细胞比例及sub-G0期细胞比例与对照组比均显著升高(P<0.01),而添加GTE24h后,G2/M期细胞比例与对照组比明显升高(P<0.05);DAPI标记示有典型的凋亡小体;经GTE(40μg/L、80μg/L)作用24h后与对照组比,SKOV3细胞中p-ERK1/2、Bcl-xl蛋白表达下调,p-ERK1/2分别为0.334±0.030、0.053±0.140(P<0.01);Bcl-xl分别为0.410±0.026、0.267±0.020(P<0.01);Caspase-3蛋白分别为0.128±0.090、0.287±0.028,与对照组相比显著升高(P<0.01),而ERK1/2蛋白则未见明显变化(P>0.05)。结论:GTE在体外能有效地抑制SKOV3细胞的生长,其作用途径可能是通过抑制ERK信号通路的转导,导致细胞周期阻滞,并下调Bcl-xl、上调Caspase-3,最终导致SKOV3细胞凋亡。
Objective: To investigate the effect of green tea extract (GTE) on the growth and proliferation of human ovarian cancer cell line SKOV3 and its mechanism of action. Methods: The proliferation and proliferation of SKOV3 cells were observed by MTT method. The morphological changes of SKOV3 cells were observed by optical microscope. The cell cycle and apoptosis of SKOV3 cells were detected by flow cytometry (FCM). The morphological changes of apoptosis were observed by DAPI. The expressions of p-ERK1 / 2, ERK1 / 2, Caspase-3 and Bcl-xl were detected by Western blotting. Results: With the increase of GTE concentration and the prolongation of action time, the survival rate of SKOV3 cells decreased gradually (P <0.01), the number of SKOV3 cells decreased gradually and the cell damage gradually increased. After treated by GTE (80μg / L) for 24h, 48h, 72h (P <0.01). The percentage of cells in S phase and the proportion of cells in sub-G0 phase were significantly higher than that in control group (P <0.01), while the percentage of cells in G0 / G1 phase was significantly lower than that in control group (P <0.05). The percentage of apoptotic cells in G2 / M phase was significantly higher than that in control group (P <0.05). The apoptotic bodies were detected by DAPI. After treated by GTE (40μg / L, 80μg / L) , The expression of p-ERK1 / 2 and Bcl-xl in SKOV3 cells was down-regulated, and the expression of p-ERK1 / 2 was 0.334 ± 0.030,0.053 ± 0.140 (P <0.01); Bcl-xl was 0.410 ± 0.026,0.267 ± 0.020 P <0.01). Caspase-3 protein was 0.128 ± 0.090,0.287 ± 0.028, which was significantly higher than that of the control group (P <0.01), while no significant change was observed in ERK1 / 2 protein (P> 0.05). CONCLUSION: GTE can effectively inhibit the growth of SKOV3 cells in vitro. The mechanism may be through inhibiting the transduction of ERK signaling pathway, leading to cell cycle arrest, down-regulating Bcl-xl and up-regulating Caspase-3, leading to the apoptosis of SKOV3 cells Death.