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目的观察盐霉素联合顺铂对人胃癌细胞MKN-45增殖和凋亡的影响并探讨其作用机制。方法体外培养人胃癌细胞MKN-45至对数生长期,采用噻唑蓝(MTT)比色法分别检测盐霉素、顺铂及联合用药时细胞的增殖情况;吖啶橙/溴化乙锭(AO/EB)凋亡试剂盒荧光染色观察细胞凋亡情况;Western blotting法检测NF-κB、caspase-3蛋白表达水平。结果 MTT法显示盐霉素、顺铂均可抑制细胞增殖(P<0.05),联合给药比单药抑制效果更为显著(P<0.05);AO/EB荧光染色可见盐霉素、顺铂分别单药给药后,与对照组相比细胞凋亡形态明显,两药联合凋亡效果更为显著;Western blotting结果显示联合组与单药组和对照组相比NF-κB蛋白表达下降(P<0.01),caspase-3蛋白表达升高(P<0.01)。结论盐霉素可抑制胃癌细胞增殖,与顺铂联合应用可增强顺铂对人胃癌细胞MKN-45的抑制作用,其机制可能为抑制NF-κB活化,上调下游凋亡基因caspase-3的表达。
Objective To observe the effect of salinomycin combined with cisplatin on proliferation and apoptosis of human gastric cancer cell line MKN-45 and to explore its mechanism. Methods Human gastric cancer cell line MKN-45 was cultured in vitro to the logarithmic growth phase. MTT assay was used to detect the proliferation of salinomycin, cisplatin and combination therapy. Acridine orange / ethidium bromide AO / EB) apoptosis kit was used to observe the cell apoptosis. The protein expression of NF-κB and caspase-3 was detected by Western blotting. Results MTT assay showed that both salinomycin and cisplatin could inhibit the cell proliferation (P <0.05), and the combination therapy was more effective than the single drug (P <0.05). The fluorescence intensity of salinomycin and cisplatin After single drug administration, compared with the control group, the morphous of apoptosis was obvious, and the combined effect of the two drugs was more significant. Western blotting showed that the expression of NF-κB decreased in the combined group compared with the single drug group and the control group P <0.01), the expression of caspase-3 protein increased (P <0.01). Conclusion Salinomycin can inhibit the proliferation of gastric cancer cells. The combination of cisplatin and cisplatin can enhance the inhibitory effect of cisplatin on human gastric cancer cell line MKN-45. The possible mechanism is that salinomycin can inhibit the activation of NF-κB and upregulate the expression of caspase-3 .