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目的探讨佛波酯(TPA)处理人胃癌MGC80-3细胞过程中磷酯酶Cγ-2(PLCγ-2)的意义。方法通过DAPI染色,荧光显微镜观察分析TPA对胃癌MGC80-3细胞的影响;借助核浆分离手段获得胃癌细胞核浆蛋白,并通过免疫印迹(Western blotting)检测TPA对PLCγ-2蛋白表达水平影响;通过免疫荧光技术处理,激光扫描共焦显微镜观察胃癌细胞内PLCγ-2蛋白的定位和转运;用PLCγ-2的抑制剂(U73122)预处理细胞,免疫印迹和激光扫描共焦显微镜分别检测其对TPA作用胃癌细胞内PLCγ-2的影响;以及荧光显微镜下观察分析U73122是否影响TPA对胃癌细胞的作用。结果TPA诱导胃癌MGC80-3细胞凋亡;同时TPA提高PLCγ-2蛋白表达水平,并诱导其发生核浆转运;其抑制剂(U73122)可以降低TPA对PLCγ-2蛋白表达水平的作用,但并不能影响TPA诱导胃癌细胞凋亡;而TPA诱导的PLCγ-2核浆转运却没有被PLCγ-2抑制剂(U73122)阻止。结论尽管TPA提高了胃癌MGC80-3细胞中PLCγ-2表达水平,但PLCγ-2表达水平和TPA诱导的胃癌MGC80-3凋亡没有直接关系,而其核浆转运可能与TPA诱导的胃癌细胞凋亡相关。
Objective To investigate the effect of phorbol ester (TPA) on phospholipase Cγ-2 (PLCγ-2) in human gastric cancer cell line MGC80-3. Methods The effect of TPA on gastric cancer cell line MGC80-3 was analyzed by DAPI staining and fluorescence microscope. The nuclear plasma protein of gastric cancer cell line was obtained by nuclear plasma separation and the effect of TPA on PLCγ-2 protein expression was detected by Western blotting. Immunofluorescence staining and laser scanning confocal microscopy were used to observe the localization and translocation of PLCγ-2 protein in gastric cancer cells. The cells were pretreated with inhibitor of PLCγ-2 (U73122) and detected by immunofluorescence and laser scanning confocal microscopy The effect of PLCγ-2 in gastric cancer cells; and observe the effect of U73122 on the effect of TPA on gastric cancer cells under the fluorescence microscope. Results TPA induced the apoptosis of gastric cancer MGC80-3 cells. At the same time, TPA increased the expression of PLCγ-2 protein and induced its nuclear transport. The inhibitor of U73122 could reduce the effect of TPA on PLCγ-2 protein expression, Can not affect TPA-induced gastric cancer cell apoptosis; and TPA-induced PLCγ-2 nuclear plasma transport was not prevented by PLCγ-2 inhibitor (U73122). Conclusion Although TPA increases the expression of PLCγ-2 in gastric cancer MGC80-3 cells, the expression level of PLCγ-2 is not directly related to TPA-induced gastric cancer MGC80-3 apoptosis, and its nuclear transport may be related to TPA-induced gastric cancer cell apoptosis Death related.