中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2012年
12期
938-943
,共6页
邢同京%徐洪涛%余文庆%姜丹凤
邢同京%徐洪濤%餘文慶%薑丹鳳
형동경%서홍도%여문경%강단봉
微小RNA-122%甲基化调控%肝细胞癌%细胞凋亡
微小RNA-122%甲基化調控%肝細胞癌%細胞凋亡
미소RNA-122%갑기화조공%간세포암%세포조망
MicroRNA-122%Methylation regulation%Hepatocellular carcinoma%Apoptosis
目的 探讨DNA甲基化对肝特异性miRNA-122表达调控以及肝癌细胞增殖和凋亡的影响.方法 甲基化测序检测肝细胞中miRNA-122启动子区的甲基化状态.实时定量PCR检测肝细胞中miRNA 122表达水平.流式细胞仪与CCK8检测去甲基化对肝癌细胞增殖和凋亡的影响.结果 与人原代正常肝细胞[(21.9±11.4)％]比较,Huh7、HepG2、QSG-7701细胞系miRNA-122启动子区的甲基化程度[分别为(87.6±9.3)％、(89.0±14.3)％、(69.5±11.5)％],均明显升高(P=0.000),尤其以Huh7、HepG2两种癌细胞系升高最为明显.与人原代正常肝细胞[(2.83×104±3746)]比较,三种细胞系miRNA-122的表达明显下降,其中以Huh7和HepG2两种癌细胞系降低最明显(P=0.007).与空白对照比较,在10 μmol/L的5氮杂-2′-脱氧胞苷作用下,Huh7和HepG2两种癌细胞系甲基化程度明显降低(P=0.038,0.025);而其miRNA-122表达明显升高(P=0.008,0.003).与空白对照组比较,Huh7细胞与HepG2细胞经去甲基化处理后细胞的凋亡均明显增加(P=0.001,0.027).结论 肝特异性miRNA-122的表达受DNA甲基化的明显调控,且与肝癌细胞的凋亡密切相关.miRNA122的甲基化调控可能参与了肝癌的发生和发展.
目的 探討DNA甲基化對肝特異性miRNA-122錶達調控以及肝癌細胞增殖和凋亡的影響.方法 甲基化測序檢測肝細胞中miRNA-122啟動子區的甲基化狀態.實時定量PCR檢測肝細胞中miRNA 122錶達水平.流式細胞儀與CCK8檢測去甲基化對肝癌細胞增殖和凋亡的影響.結果 與人原代正常肝細胞[(21.9±11.4)％]比較,Huh7、HepG2、QSG-7701細胞繫miRNA-122啟動子區的甲基化程度[分彆為(87.6±9.3)％、(89.0±14.3)％、(69.5±11.5)％],均明顯升高(P=0.000),尤其以Huh7、HepG2兩種癌細胞繫升高最為明顯.與人原代正常肝細胞[(2.83×104±3746)]比較,三種細胞繫miRNA-122的錶達明顯下降,其中以Huh7和HepG2兩種癌細胞繫降低最明顯(P=0.007).與空白對照比較,在10 μmol/L的5氮雜-2′-脫氧胞苷作用下,Huh7和HepG2兩種癌細胞繫甲基化程度明顯降低(P=0.038,0.025);而其miRNA-122錶達明顯升高(P=0.008,0.003).與空白對照組比較,Huh7細胞與HepG2細胞經去甲基化處理後細胞的凋亡均明顯增加(P=0.001,0.027).結論 肝特異性miRNA-122的錶達受DNA甲基化的明顯調控,且與肝癌細胞的凋亡密切相關.miRNA122的甲基化調控可能參與瞭肝癌的髮生和髮展.
목적 탐토DNA갑기화대간특이성miRNA-122표체조공이급간암세포증식화조망적영향.방법 갑기화측서검측간세포중miRNA-122계동자구적갑기화상태.실시정량PCR검측간세포중miRNA 122표체수평.류식세포의여CCK8검측거갑기화대간암세포증식화조망적영향.결과 여인원대정상간세포[(21.9±11.4)％]비교,Huh7、HepG2、QSG-7701세포계miRNA-122계동자구적갑기화정도[분별위(87.6±9.3)％、(89.0±14.3)％、(69.5±11.5)％],균명현승고(P=0.000),우기이Huh7、HepG2량충암세포계승고최위명현.여인원대정상간세포[(2.83×104±3746)]비교,삼충세포계miRNA-122적표체명현하강,기중이Huh7화HepG2량충암세포계강저최명현(P=0.007).여공백대조비교,재10 μmol/L적5담잡-2′-탈양포감작용하,Huh7화HepG2량충암세포계갑기화정도명현강저(P=0.038,0.025);이기miRNA-122표체명현승고(P=0.008,0.003).여공백대조조비교,Huh7세포여HepG2세포경거갑기화처리후세포적조망균명현증가(P=0.001,0.027).결론 간특이성miRNA-122적표체수DNA갑기화적명현조공,차여간암세포적조망밀절상관.miRNA122적갑기화조공가능삼여료간암적발생화발전.
Objective miRNA-122 levels may correlate with liver cancer prognosis,and therefore understanding its expression is crucial for future treatment.This study investigates the effect of DNA methylation on the expression of liver specific miRNA-122 and its effects on proliferation and apoptosis of hepatocellular carcinoma cells.Methods Methylation sequencing detected the methylation of the miRNA-122 promoter region,and the level of miRNA-122 expression was measured by using real-time quantitative PCR.The proliferation and apoptosis of hepatocellular cell lines were detected by flow cytometry and CCK8.Results Compared with human primary hepatocytes [(21.9 ± 11.4)％],the level of miRNA-122 promoter methylation in Huh7,HepG2,and QSG-7701 cell lines were (87.6±9.3) ％,(89.0 ± 14.3)％,and (69.5 ±11.5)％,respectively.This represents a significant increase (P=0.000),especially in Huh7 and HepG2 cell lines.Compared with human primary hepatocytes (2.83× 104 ±3746),the levels of miR-122 expression in the above three cell lines were significantly decreased,especially in Huh7 and HepG2 cell lines (P=0.007).After treatment with 5-Aza-dc,the degree of methylation in Huh7 and HepG2 cell lines were significantly lower than that of the blank group (P=0.038,P=0.025),and the levels of miRNA-122 expression were significantly elevated (P=0.008,P=0.003).Also,compared with the control groups,the apoptosis of Huh7 cells and HepG2 cells were significantly increased (P=0.001,0.027).Conclusion The expression of miRNA-122 is regulated by DNA methylation and correlated with the apoptosis of liver cancer cells.Therfore,the methylation regulation of miRNA-122 expression might be involved in the occurrence and development of hepatocellular carcinoma.
