腺苷干预人结直肠癌SW480细胞RECK基因甲基化及其机制

发布时间：2018-08-08 20:08

【摘要】：目的探讨腺苷干预人结直肠癌SW480细胞RECK基因甲基化及其机制。方法分别用0、3.0 mmol/L的腺苷干预SW480细胞72 h后,亚硫酸氢盐测序法(BSP)检测RECK基因启动子区Cp G岛甲基化情况;逆转录PCR、Western blot、流式细胞仪检测腺苷处理前后RECK基因mRNA和蛋白的表达、甲基转移酶(methyltransferases DNMT1,DNMT3a)蛋白表达及细胞凋亡的变化。结果腺苷干预后RECK基因甲基化程度明显低于对照组[(49.14±2.38)%vs(74.84±2.38)%,P0.01];腺苷干预后RECK基因mRNA转录及蛋白表达、细胞凋亡均明显高于对照组[0.335 3±0.350 2 vs 0.080 0±0.003 6,0.603 0±0.017 8 vs 0.090 0±0.003 0;(27.90±2.32)%vs(5.09±0.30)%,P0.01];甲基转移酶(DNMT1、DNMT3a)蛋白表达均明显低于对照组(0.141 3±0.002 1 vs 0.645 0±0.009 2,0.130 0±0.004 6 vs 0.526 7±0.009 5,P0.01)。结论腺苷通过抑制甲基转移酶的活性,降低RECK基因DNA启动子区域的甲基化水平,使RECK基因mRNA及蛋白表达水平增加,促使SW480细胞凋亡。
[Abstract]:Objective to investigate the effect of adenosine on methylation of RECK gene in human colorectal cancer SW480 cells and its mechanism. Methods after SW480 cells were treated with adenosine for 72 h, methylation of RECK gene promoter was detected by (BSP), reverse transcriptase polymerase chain reaction (RT-PCR) and flow cytometry were used to detect the expression of mRNA and protein of RECK gene before and after adenosine treatment. Expression of methyltransferase (methyltransferases DNMT1, DNMT3a) and apoptosis. Results the methylation degree of RECK gene after adenosine intervention was significantly lower than that in the control group [(49.14 卤2.38) vs (74.84 卤2.38) P 0.01], and the mRNA transcription and protein expression of RECK gene after adenosine intervention were significantly lower than those in the control group (49.14 卤2.38). Apoptosis was significantly higher than that in the control group [0.335 3 卤0.350 2 vs 0.080 0 卤0.003 6 卤0.6030 卤0.017 8 vs 0.090 0 卤0.003 0; (27.90 卤2.32) vs (5.09 卤0.30) P 0.01], and the expression of DNMT1 + DNMT3a protein was significantly lower than that in the control group (0.141 3 卤0.002 1 vs 0.645 0 卤0.009 2 卤0.130 0 卤0.004 6 vs 0.526 7 卤0.009 5P0.01), and the expression of DNMT1a protein was significantly lower than that in the control group (0.141 3 卤0.002 1 vs 0.645 0 卤0.009 2 卤0.004 6 vs 0.526 7 卤0.009 5P0.01). Conclusion adenosine can inhibit the activity of methyltransferase, decrease the methylation level of DNA promoter of RECK gene, increase the expression of mRNA and protein of RECK gene, and promote the apoptosis of SW480 cells.
【作者单位】：重庆医科大学附属第二临床学院胃肠外科;美国俄勒冈州Legacy研究所;
【基金】：重庆市自然科学基金(CSTC2012jja10063)~~
【分类号】：R735.34

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