氧化槐果碱对胃癌MGC80-3细胞增殖和凋亡的影响

Effects of Oxysophocarpine on the Proliferation and Apoptosis of Gastric Cancer MGC80-3 Cells

关键词:  氧化槐果碱  胃癌  增殖  凋亡

Key words:  oxysophocarpine  gastric cancer  proliferation  apoptosis

摘要:

目的 观察氧化槐果碱对胃癌MGC80-3细胞增殖和凋亡的影响及机制。方法 采用CCK-8、Hoechest 33342染色、倒置显微镜和流式细胞术测定氧化槐果碱干预后对细胞的增殖、凋亡的影响，qRT-PCR检测胃癌MGC80-3细胞中Bcl-2、Bax mRNA水平，Western blot法检测胃癌MGC80-3细胞中Bcl-2、Bax、cleaved caspase-3、cleaved caspase-9蛋白水平。结果 氧化槐果碱能抑制胃癌MGC80-3细胞增殖（P<0.05），并具有时间和浓度依赖性。0.48，0.95，1.91 mmol·L^-1氧化槐果碱能诱导细胞凋亡（P<0.05），凋亡抑制剂Z-DEVD-FMK能逆转氧化槐果碱的凋亡诱导作用，0.95，1.91 mmol·L^-1氧化槐果碱能上调Bax的mRNA水平，下调Bcl-2的mRNA水平，差异均有统计学意义（P<0.05）。0.95，1.91 mmol·L^-1氧化槐果碱能上调Bax、cleaved caspase-3、cleaved caspase-9的蛋白水平，下调Bcl-2蛋白水平，差异均有统计学意义（P<0.05或P<0.01）。凋亡抑制剂Z-DEVD-FMK能逆转氧化槐果碱对Bcl-2、Bax、cleaved caspase-3、cleaved caspase-9蛋白的影响。结论 氧化槐果碱能通过调节Bcl-2、Bax、cleaved caspase-3、cleaved caspase-9的表达水平，抑制胃癌MGC80-3细胞增殖，诱导细胞凋亡。

Abstract:

OBJECTIVE To observe the effect of oxysophocarpine on the proliferation and apoptosis of gastric cancer MGC80-3 cells and its mechanism. METHODS CCK-8, Hoechest 33342 staining, inverted microscope and flow cytometry were used to determine the effect of oxidized saponin on cell proliferation and apoptosis. qRT-PCR was used to detect Bcl-2 and Bax mRNA levels in gastric cancer MGC80-3 cells. Western blot was used to detect the protein levels of Bcl-2, Bax, cleaved caspase-3 and cleaved caspase-9 in gastric cancer MGC80-3 cells. RESULTS Oxysophocarpine effectively inhibited the proliferation of gastric cancer MGC80-3 cells in a time-and dose-dependent manner(P<0.05). 0.48, 0.95, 1.91 mmol·L^-1 oxysophocarpine could induce apoptosis(P<0.05). Apoptosis inhibitor Z-DEVD-FMK could reverse the apoptosis-inducing effect of oxysophocarpine. 0.95, 1.91 mmol·L^-1 oxysophocarpine could up-regulate the mRNA level of Bax and down-regulate the mRNA level of Bcl-2, the differences were statistically significant(P<0.05). 0.95 mmol·L^-1 and 1.91 mmol·L^-1 oxysophocarpine could up-regulated the protein levels of Bax, cleaved caspase-3 and cleaved caspase-9, and down-regulated the level of Bcl-2 protein, the differences were statistically significant(P<0.05 or P<0.01). The inhibitor of apoptosis Z-DEVD-FMK reversed the effects of oxysophocarpine on Bcl-2, Bax, cleaved caspase-3 and cleaved caspase-9 proteins. CONCLUSION Oxysophocarpine can inhibit the proliferation of gastric cancer MGC80-3 cells and induce apoptosis by regulating the expression of Bcl-2, Bax, cleaved caspase-3 and cleaved caspase-9.