Inhibitory Effects Of Oxaliplatin Combined With Decitabine On GC Cell Line MKN-28 Via Wnt/beta-catenin Signaling Pathway

Objectives: This study intends to analyze the inhibition of oxaliplatin combined with decitabine in the proliferation of well-differentiated gastric cancer(GC)cell line(MKN-28)through regulating Wnt/beta-Catenin signaling pathway.Methods: MKN-28 cells were randomly divided into the control group(n = 6),the oxaliplatin group(n = 6),the decitabine group(n = 6),the oxaliplatin combined with decitabine(oxaliplatin + decitabine)group(n = 6)and the Wnt/beta-Catenin signaling pathway inhibitor XAV-939 group(the XAV-939 group,n = 6).The control group was treated with DMSO,the oxaliplatin group was treated with oxaliplatin(10mg/L,100?L),the decitabine group was treated with decitabine(20mg/L,100?L),the oxaliplatin + decitabine group was treated with both oxaliplatin(10mg/L,100?L)and decitabine(20mg/L,100?L),and the XAV-939 group was treated with XAV-939(10mg/L,100?L).MTT assay was applied to analyze the MKN-28 cell proliferation.DAPI staining was exhibited to detect the MKN-28 cell apoptosis.Real-time PCR(RT-PCR)was used to analyze the mRNA expressions of apoptosis proteins caspase-3 and caspase-6.The Wnt/beta-Catenin signaling pathway proteins expressions were measured using western Blotting and immunocytochemistry.Results: Compared with the control group,the inhibition rates of MKN-28 cells were significantly higher at 1 h,2 h and 4 h in the oxaliplatin,decitabine,oxaliplatin + decitabine and XAV-939 groups(P < 0.05).Moreover,the inhibition rates of MKN-28 cells were significantly increased at 1 h,2 h and 4 h in the oxaliplatin + decitabine group when compared with the oxaliplatin,decitabine and XAV-939 groups(P < 0.05).In comparison to the control group,the number of apoptotic corpuscles,and mRNA expressions of caspase-3 and caspase-6 were obviously higher in the other four groups(P < 0.05),which were the highest in the oxaliplatin + decitabine group.Wnt and beta-Catenin proteins were mainly localized in the cytoplasm.Wnt and ?-Catenin protein expressions and fluorescence intensity were significantly lower in the other four groups than those were in the control group(P < 0.05).Meanwhile,the oxaliplatin + decitabine group showed the lowest Wnt and ?-Catenin protein expressions and fluorescence intensity.Conclusion: This study reveals that oxaliplatin combined with decitabine can inhibit protein expressions of Wnt/beta-Catenin signaling pathway and promote protein expressions of caspase 3 and caspase 6,thus downregulating the MKN-28 cells proliferation and upregulating the MKN-28 cells apoptosis in a time-dependent manner.