| Objective:To investigate the influence of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the proliferation and apoptosis of human renal carcinoma cell line OS-RC-2,and to study the influence of 5-Aza-CdR on methylati on ofγ-catenin gene,so as to provide insights into origin of renal carcinoma and the possibility of its application in clinical treatment.Methods:Human renal carcinoma OS-RC-2 cells were treated with10-7, 10-6,10-5,10-4mol/L 5-Aza-CdR,respectively.Consequently,Then the morphological pattern changes were observed under invert-microscope and transmissional electron microscope.The growth rate of the cells was detected by MTT assay.The apoptosis was analyzed by flowcytometry. The immunocytochemistry and western-blot methods were used to detectγ-catenin protein levels in the cell line before and after treatment with 5-Aza-CdR.The methylation and unmethylaion status ofγ-catenin gene were detected by methylation special PCR(MSP).Results:The apoptotic morphology was observed,by invert-microscope and transmiting electron microscope,5-Aza-CdR inhibited the proliferateion of OS-RC-2 cells in a time and concentration -dependent manner(P＜0.05),there were significant increases in apoptotic rates of OS-RC-2 cells[(3.74±0.34)%,(7.85±0.59)%,(12.93±1.32)%vs(0.86±0.08)%,F=189. 7,P＜0.01]after 5-Aza-CdR treatment in comparision with control group. The immunocytochemistry and western blot indicated that 5-Aza-CdR could re-express theγ-catenin proteins after treatment with 5-Aza-CdR,and the 5-Aza-CdR treatment also increase the expression levels ofγ-catenin protein in a concentration-dependent manne (P＜0.05).The methylation status ofγ-catenin gene promoter region was reversed with 10-5 mol/L5-Aza-CdR for 72h.Conclusions:5-Aza-CdR may effectively cause the demethylation ofγ-catenin gene CpG-rich promoter regions,and recover theγ-catenin protein expression,so as to inhibit the growth of OS-RC-2 cells and induce cell apoptosis. |
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