CAJ | 학술논문

目的初步探讨表没食子儿茶素没食子酸酯(EGCG)对人结肠腺癌细胞株COLO 320 Alu、LINE-1序列甲基化的影响,探讨EGCG在改变COLO 320抑癌基因p16高甲基化的同时,是否会改变全基因组的甲基化模式.方法甲基特异性PCR(MSP)验证20μmol/L的EGCG干预培养COLO 320后p16启动子去甲基化作用,亚硫酸氢盐修饰后测序法(BSP)检测不同浓度EGCG干预培养COLO 320后Alu、LINE-1序列甲基化水平.结果 20μmol/L的EGCG对高甲基化的p16启动子去甲基化作用具有统计学意义(P =0.000),但实验所设的EGCG各浓度对Alu、LINE-1序列甲基化水平影响无统计学意义(Alu:P=0.794,LINE-1:P=0.836).结论 EGCG在改变肿瘤细胞株特异抑癌基因高甲基化模式的同时,能够维持全基因组的甲基化水平,保持基因组的稳定性.
목적 초보탐토표몰식자인다소몰식자산지(EGCG)대인결장선암세포주COLO 320 Alu、LINE-1서렬갑기화적영향,탐토EGCG재개변COLO 320억암기인p16고갑기화적동시,시부회개변전기인조적갑기화모식.방법 갑기특이성PCR(MSP)험증20μmol/L적EGCG간예배양COLO 320후p16계동자거갑기화작용,아류산경염수식후측서법(BSP)검측불동농도EGCG간예배양COLO 320후Alu、LINE-1서렬갑기화수평.결과 20μmol/L적EGCG대고갑기화적p16계동자거갑기화작용구유통계학의의(P =0.000),단실험소설적EGCG각농도대Alu、LINE-1서렬갑기화수평영향무통계학의의(Alu:P=0.794,LINE-1:P=0.836).결론 EGCG재개변종류세포주특이억암기인고갑기화모식적동시,능구유지전기인조적갑기화수평,보지기인조적은정성.
Objective To study the effect of Epigallocatechin-3-gallate (EGCG) on DNA methylation of Alu and LINE-1 elements in human colon carcinoma cell line 320 (COLO 320) in vitro,and discuss if EGCG would change the whole genome methylation patterns besides changing the tumor-suppressor gene p16 high methylation patterns.Methods In vitro,Methylation-Specific PCR,MS-PCR (MSP) was used to verify demethylation of high methylation suppressor gene p16 promoter in COLO 320 after intervention by EGCG of 20 μmol/L,Bisulfite Sequencing PCR (BSP) was employed to detect the methylation levels of Alu and LINE-1 elements in COLO 320 after intervention in different concentration EGCG.Results EGCG of 20 μmol/L had effect of demethylaiton to high methylation suppressor gene p16 promoter (P =0.000),while different concentrations of EGCG had no significant impact on methylation levels of Alu and LINE-1 elements (Alu:P =O.794,LINE-1:P =0.836).Conclusion EGCG can maintain the whole genome methylation level and guarantee the stability of the genome besides changing high methylation patterns of tumor-suppressor genes.