【摘要】 目的分析奥沙利铂联合地西他滨抑制高分化胃癌细胞株MKN-28作用的主要机制及Wnt/β-Catenin信号转导通路的作用。方法体外常规培养胃癌MKN-28MKN45细胞,将细胞随机分为5组:对照组(DMSO)、奥沙利铂组(奥沙利铂10 mg/L)、地西他滨组(地西他滨20 mg/L)、奥沙利铂联合地西他滨组(奥沙利铂10 mg/L+地西他滨20 mg/L)、抑制剂XAV-939组(XAV-939 10 mg/L)。MTT法分析各组胃癌细胞经处理后的增殖抑制作用,DAPI染色法分析各组细胞的凋亡,Western Blotting(WB)法和免疫细胞化学法分析Wnt/β-Catenin信号通路蛋白的变化。结果与对照组相比,奥沙利铂组、地西他滨组、联合组和XAV-939组在1、2和4 h的细胞抑制率明显升高,呈时间依赖性,且差异有统计学意义(P <0.05);同时与奥沙利铂组、地西他滨组和XAV-939组相比,联合组在1、2和4 h的细胞抑制率明显升高,差异有统计学意义(P <0.05);DAPI染色分析得出,与对照组细胞相比,其余4组细胞的凋亡小体数量明显增多,且联合组细胞的凋亡小体数量最多;WB法得出,与对照组相比,其余4组细胞的Wnt和β-Catenin蛋白水平明显降低(P <0.05),细胞浆内蛋白荧光强度减弱,且奥沙利铂联合地西他滨联合组细胞的上述蛋白表达降低最明显,细胞浆内蛋白荧光强度减弱也最明显(P <0.05)。结论奥沙利铂联合地西他滨能明显抑制胃癌细胞株MKN-28增殖并呈时间依赖性,可诱导细胞凋亡。其机制与抑制Wnt/β-Catenin信号通路Wnt和β-Catenin蛋白表达有关。更多还原

【Abstract】 Objective To investigate the inhibition effect and mechanism of oxaliplatin combined withdecitabine in the inhibition of gastric cancer MKN28 cells by regulating Wnt/beta-Catenin signaling pathway.Methods The experiment was randomly divided into control group(DMSO),oxaliplatin group(10 mg/L),decitabinegroup(20 mg/L),oxaliplatin combined with decitabine group(both concentration points),Wnt/beta-Cateninsignaling pathway inhibitor XAV-939 group(10 mg/L). Gastric cancer MKN28 cell was treated with 1,2,4 h forsubsequent follow-up. MTT assay was used to detect the cell viability of the inhibition rate of cell proliferation.DAPI staining was used to analyze the cell apoptosis. Western blotting was used to analyze the expression of proteinssuch as Wnt and β-Catenin. Immunocytochemistry was used to analyze the fluorescence intensity of Wnt andβ-Catenin. Results Compared with control group,the proliferation of cells in oxaliplatin group,decitabinegroup,oxaliplatin combined with decitabine group,XAV-939 group was significantly inhibited after treatment for1,2 and 4 h with a time-dependent way(P < 0.05). DAPI staining showed that the number of apoptotic corpusclesin the other four groups was significantly higher than that in the control group,and the number of apoptotic corpus-cles in the combined group was the largest(P < 0.05). Compared with the control group,the expressions of Wnt andβ-Catenin was significantly decreased in the other four groups,and the fluorescence intensity of Wnt and beta-Catenin proteins in the cytoplasm was weaker(P < 0.05). This change is most significant in the combined group(P < 0.05). Conclusion Oxaliplatin combined with decitabine can inhibit the proliferation of gastric cancer cellsand induce apoptosis with a time-dependent way by inhibiting Wnt/beta-Catenin signaling pathway.更多还原