硫酸右旋糖苷对人胃癌MGC-803细胞增殖和凋亡的影响

Effects of dextran sulfate on proliferation and apoptosis of human gastric cancer MGC-803 cells

目的探讨硫酸右旋糖苷(DS)对人胃癌MGC-803细胞增殖、集落形成和凋亡的影响及可能的机制。方法培养人胃癌MGC-803细胞,分为对照组(PBS处理)和实验组(0.3%DS处理);应用EdU细胞增殖实验检测DS对胃癌MGC-803细胞增殖能力的影响;平板克隆形成实验检测DS对胃癌MGC-803细胞集落形成能力的影响;AnnexinⅤ-PI双染流式检测细胞凋亡的变化;蛋白质印迹法(Western blot)检测细胞不同时间点(2、 8、 12、 24 h)EZH2、 Cleaved-caspase3蛋白表达水平。结果 EdU实验结果显示,与对照组相比,DS明显抑制胃癌MGC-803细胞增殖(P<0.01)。平板克隆形成实验结果表明,DS组集落形成能力较对照组显著降低(P<0.01), AnnexinⅤ-PI双染流式结果显示,实验组细胞凋亡率明显高于对照组(P<0 .01);Westernblot结果表明,实验组EZH2的表达较对照组明显下调(P<0 .05), Cleaved-caspase3的表达较对照组明显上调(P<0 .05)。结论 DS能明显抑制人胃癌MGC-803细胞增殖,诱导凋亡,其机制可能与抑制EZH2的表达有关。

Objective To investigate the effect of dextran sulfate (DS) on the proliferation and apoptosis in human gastric cancer MGC-803 cells and its mechanisms. Methods Human gastric cancer MGC-803 cells were cultured and divided into control group (PBS group) and experimental group (DS group). The effect of DS on the proliferation of gastric cancer MGC-803 cells was detected by EdU cell proliferation assay. Plate colony formation assay was used to detect the effect of DS on colony forming ability of MGC-803 cells. Changes of the apoptosis were determined by AnnexinV-PI double staining. Western blot assay was also used to analyze the protein expression levels of EZH2, Cleaved-caspase3 at different time points (2 h, 8 h, 12 h and 24 h). Results EdU results showed that DS significantly inhibited the proliferation of gastric cancer MGC-803 cells compared with that of the control group (P<0 .01). The plate colony formation experiment showed that cell colony forming ability was significantly decreased in experimental group than that of the control group (P<0.01). The result of AnnexinV-PI double staining showed that the apoptosis rate was significantly higher in the experimental group than that of the control group (P<0.01). Western blot analysis showed that after DS intervention the protein expression of EZH2 was significantly decreased in MGC-803 cells compared with that of control (P<0 .05), and the protein expression level of Cleaved-caspase3 was significantly increased (P<0.05). Conclusion DS can significantly inhibit the proliferation and induce apoptosis of human gastric cancer MGC-803 cells, which may be related to the inhibition of EZH2 expression.