CYP2D6基因在结直肠癌细胞侵袭和迁移中的作用及调控机制研究  

Effect of CYP2D6 in migration and invasion of colorectal cancer cells and its mechanism

作　　者：黄明 江明凤[1] 邱黎清 HUANG Ming;JIANG Mingfeng;QIU Liqing(Authors'address:Department of Clinical Laboratory,Hangzhou Cancer Hospital,Hangzhou 310002,China)

机构地区：[1]杭州市肿瘤医院检验科,310002 [2]杭州市肿瘤医院肿瘤研究所,310002

出　　处：《浙江医学》2022年第11期1139-1144,I0005,共7页Zhejiang Medical Journal

基　　金：浙江省公益技术应用社会发展项目(LGF19H310001)。

摘　　要：目的探讨细胞色素P450家族成员CYP2D6基因在结直肠癌(CRC)细胞侵袭和迁移中的作用及其调控机制。方法选取2018年6月至2019年7月杭州市肿瘤医院接受手术CRC患者的CRC组织及其配对的正常组织28对;以HT29和SW480 CRC细胞系作为研究对象,利用DNA甲基化酶抑制剂5-Aza-2'-deoxycytidine(DAC)和组蛋白去乙酰化酶抑制剂Vorinostat(SAHA)分别抑制胞内DNA甲基化和组蛋白去乙酰化,采用荧光定量PCR检测CYP2D6基因的表达水平变化,Western blot法检测细胞CYP2D6蛋白表达水平的差异,Transwell细胞侵袭实验和划痕实验检测CYP2D6基因表达水平的变化对CRC细胞侵袭和迁移能力的影响。结果DAC可有效促进HT29和SW480细胞中CYP2D6基因表达水平,而SAHA对CYP2D6基因表达水平无明显影响,两者联合使用时可协同增强HT29和SW480细胞CYP2D6基因的表达水平。Transwell细胞侵袭实验和划痕实验表明,CYP2D6基因的过表达可显著降低HT29细胞的侵袭能力和迁移能力。结论CYP2D6基因表达受表观遗传修饰的调控,相较于抑制组蛋白去乙酰化,抑制DNA的甲基化可显著上调CYP2D6转录,抑制组蛋白去乙酰化则可以发挥协同效应,共同增强细胞内CYP2D6的表达水平,CYP2D6为抑癌基因,可降低结直肠癌细胞的侵袭力和迁移力。Objective To investigate the effect of CYP2D6 in invasion and migration of colorectal cancer(CRC)cells and its mechanism.Methods Twenty-eight pairs of CRC tissues and matched normal tissues of CRC patients undergoing surgery in Hangzhou Cancer Hospital from June 2018 to July 2019 were selected.Human CRC HT29 and SW480 cells were treated with decitabine and vorinostat to inhibit the DNA methylation and histone deacetylation,respectively.The mRNA and protein expression levels of CYP2D6 in CRC cells was detected with fluorescent quantitative RT-PCR and Western blot,respectively.The invasion and migration of HT29 cells were detected by Transwell and wounding heal assays,respectively.Results The decitabine-induced DNA methylation inhibition up-regulated the expression levels of CYP2D6 in HT29 and SW480 cells.The inhibition of deacetylation of histone alone had no significant effect on CYP2D6 expression,while treatment with decitabine and vorinostat synergistically enhanced the expression of CYP2D6 in HT29 and SW480 cells.Transwell and wounding heal assays exhibited that the up-regulated CYP2D6 significantly inhibited the invasion and migration of HT29 cells.Conclusion The inhibition of DNA methylation can significantly up-regulate the transcription of CYP2D6,while inhibiting of DNA methylation and histone deacetylation can exert a synergistic effect enhancing the expression of CYP2D6 in CRC cells.The results suggested that CYP2D6 may function as a tumor suppressor gene to reduce the invasion and migration of colorectal cancer cells.

关 键 词：结直肠癌 CYP2D6 细胞侵袭 细胞迁移 表观遗传调控 

分 类 号：R735.34[医药卫生—肿瘤]