姜黄素对MDA-MB-361乳腺癌细胞DLC1基因表达影响及其机制探讨  

Effect of curcumin on DLC1 gene expression and relevant mechanism in human breast cancer cell MDA-MB-361

作　　者：刘宇飞[1,2] 王飞[1,2] 罗代珍[1,2] 蔡玲玲[1,2] 陈洪雷[3] 何钒[1,2] 王晓璐[1,2] 

机构地区：[1]三峡大学病理学研究所,湖北宜昌443000 [2]宜昌市中心人民医院病理科,湖北宜昌443003 [3]武汉大学基础医学院病理学教研室,湖北武汉430071

出　　处：《中华肿瘤防治杂志》2016年第7期426-430,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：宜昌市医疗卫生科技计划(A01301-15);宜昌市中心人民医院科研发展基金(KFJ2011004)

摘　　要：目的 DLC1下调与多种肿瘤的发生相关,DLC1基因启动子区CpG岛甲基化可能是其基因表达下调或缺失的重要机制。本研究探讨姜黄素对MDA-MB-361人乳腺癌细胞株DLC1基因表达的影响及其机制。方法应用甲基化特异性聚合酶链反应(methylation specific polymerase chain reaction,MSP)、逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)及量子点免疫荧光细胞化学(quantum dots immunofluorescence cytochemistry,QDICC)检测不同浓度姜黄素(10、20、40μmol/L)处理乳腺癌细胞株MDA-MB-361前后DLC1基因启动子甲基化状态、DLC1mRNA及蛋白表达水平变化。结果 MDA-MB-361细胞DLC1基因启动子区CpG岛呈甲基化状态,10μmol/L姜黄素无去甲基化作用,无DLC1mRNA和蛋白表达。经过20和40μmol/L姜黄素处理后,DLC1基因启动子区呈现一定程度的去甲基化,其mRNA和蛋白表达增加,mRNA相对表达量分别为0.88±0.14和1.31±0.11,DLC1蛋白均(++)。不同浓度姜黄素处理组间差异有统计学意义,F=157.344,P<0.001。40μmol/L姜黄素较20μmol/L姜黄素处理细胞后mRNA和蛋白表达稍增加,但差异无统计学意义,F=16.325,P=0.016。结论 MDA-MB-361乳腺癌细胞中DLC1基因表达沉默与启动子甲基化有关,20和40μmol/L姜黄素能部分逆转该细胞的DLC1基因甲基化状态,恢复该基因表达。OBJECTIVE This study was to investigate the effect of curcumin on expression and relevant mechanism of tumor suppressor gene DLC1 in human breast cancer cell MDA-MB-361. METHODS MDA-MB-361 cells were treated by different concentrations of curcumin（10, 20, 40μmol/L）. Methyiation-specific PCR （MSP） was used to examine the methylation status of DLC1 promoter by treatment of curcumin. DLC1 mRNA levels and DLC1 protein were examined by reverse transcription PCR （RT-PCR） and quantum dots immunofluorescenee cytochemistry （QDICC） respectively. RE- SULTS There was methylation in DLC1 promoter of MDA-MB-361 cell. 10 μmol/L curcumin has no demethylation effect and can not induce the expression of DLC1 mRNA and protein in MDA-MB-361 cell. The level of promotor methyl ation decreased notably and the expression of DLC1 mRNA and protein in MDA-MB-361 cell increased after treated by 20 μmol/L and 40 μmol/L curcumin. The relative mRNA level were 0. 88 ± 0. 14 and 1. 31 ± 0. 11, respectively. The DLC1 protein showed medium positive degree in all two groups. There were significant differences in different concentrations of cur cumin treatment group （ F=-157. 344, P 〈 0.001）. The tuRN A and protein expression levels also tr ended to increase after treated by 40 μmol/L curumin compared with 20 μmol/L curumin, but no statistical difference were observed （F=16. 325, P=0. 016）. CONCLUSION Methylation of promoter may be one of the important inactivating factors of DLC1 gene in MDA-MB-361 human breast cancer cells, and the 20 μmol/L and 40μmol/L cureumin can partially reverse this methylation and recover the expresion of DLC1 gene.

关 键 词：DLC1基因 启动子甲基化 乳腺肿瘤 姜黄素 

分 类 号：R737.9[医药卫生—肿瘤]