高压氧联合紫杉醇对人胃癌AGS细胞周期及活性氧水平的影响

Effects of hyperbaric oxygen combined with paclitaxel on cell cycle and ROS level of lung cancer AGS cells

目的探讨高压氧(HBO)联合紫杉醇(PTX)对体外培养的人胃癌AGS细胞株增殖、活性氧水平(ROS)及细胞周期的影响。方法体外培养人胃癌AGS细胞完成后,将细胞分为4组:对照组、PTX组、HBO组和HBO+PTX组。采用CCK-8法检测HBO和PTX联用对胃癌AGS细胞增殖的抑制作用,流式细胞术检测细胞内ROS水平和细胞周期阻滞情况,Western blotting实验检测AGS细胞内细胞周期相关蛋白的表达水平。结果HBO+PTX组AGS细胞存活率明显低于对照组和HBO组(P<0.01),也低于PTX组(P<0.05);HBO+PTX组AGS细胞内ROS水平明显高于对照组和HBO组(P<0.01),也高于PTX组(P<0.05)。PTX将AGS细胞周期阻滞在S期(P<0.05),同时发现HBO+PTX组S期占比略高于PTX组(P>0.05)。HBO+PTX组Cyclin A1表达量明显低于其他3组(P<0.05或P<0.01),CDK2表达量低于对照组和HBO组(P<0.05),略低于PTX组(P>0.05)。结论HBO能够有效增强PTX对AGS细胞的增殖抑制作用,提高AGS细胞内ROS水平,并阻滞细胞周期,从而抑制AGS细胞的分裂与增殖。

Objective To explore the effect of hyperbaric oxygen(HBO)combined with paclitaxel(PTX)on the proliferation,level of reactive oxygen species(ROS),and cell cycle of human gastric cancer AGS cell line cultured in vitro.Methods After the completion of in vitro culture,the AGS cells were divided into four groups,including control group,PTX group,HBO group,and HBO+PTX group.Cell Counting Kit-8(CCK-8)assay was used to detect the inhibitory effect of HBO combined with PTX on the proliferation of AGS cells.Flow cytometry was adopted to detect the intracellular ROS level and cell cycle arrest.Western blotting was used to assess the expression levels of cell cycle-related proteins.Results The survival rate of AGS cells in the HBO+PTX group was significantly lower than those in the control group and the HBO group(P<0.01),and also lower than that in the PTX group(P<0.05).The ROS level of AGS cells in the HBO+PTX group was significantly higher than those in the control group and the HBO group(P<0.01),and also higher than that in the PTX group(P<0.05).PTX blocked the AGS cell cycle in S phase(P<0.05),and the percentage of cells at S phase in the HBO+PTX group was found slightly higher than that in the PTX group(P>0.05).The expression level of Cyclin A1 in the HBO+PTX group was significantly lower than those in the other three groups(P<0.05 or P<0.01);the expression level of CDK2 in the HBO+PTX group was lower than those in the control group and the HBO group(P<0.05),and also slightly lower than that in the PTX group(P>0.05).Conclusion HBO can effectively enhance the inhibitory effect of PTX on the proliferation of AGS cells,increase the intracellular ROS level in AGS cells,and block the cell cycle,thereby inhibiting the differentiation and proliferation of AGS cells.