[1]王 虹,李丽丽,张吉才,等.5-氮杂-2'脱氧胞苷对肺癌A549/DDP细胞hMLHl,MGMT基因甲基化及其表达的影响[J].现代检验医学杂志,2015,30(03):83-86.[doi:10.3969/j.issn.1671-7414.2015.03.024]
　WANG Hong,LI Li-li,ZHANG Ji-cai,et al.Effects of 5-Aza-Cde on DNA Methylation and Expression of hMLHl and MGMT Gene in Lung Cancer Cell Line A549/DDP[J].Journal of Modern Laboratory Medicine,2015,30(03):83-86.[doi:10.3969/j.issn.1671-7414.2015.03.024]
点击复制

5-氮杂-2'脱氧胞苷对肺癌A549/DDP细胞hMLHl,MGMT基因甲基化及其表达的影响()
分享到：

《现代检验医学杂志》[ISSN:/CN:]

卷:
第30卷
期数:
2015年03期
页码:
83-86
栏目:
论著
出版日期:
2015-06-10

文章信息/Info

Title:
Effects of 5-Aza-Cde on DNA Methylation and Expression of hMLHl and MGMT Gene in Lung Cancer Cell Line A549/DDP
文章编号:
1671-7414(2015)03-083-04
作者:
王 虹1; 李丽丽1; 张吉才2; 高 波2; 骆海军1
1.十堰市中医医院检验科,湖北十堰 442012;
2.湖北医药学院附属太和医院检验科,湖北十堰 442000
Author(s):
WANG Hong1; LI Li-li1; ZHANG Ji-cai2; GAO Bo2; LUO Hai-jun1
1.Department of Chinical Laboratory,Shiyan Hospital of Traditional Chinese Medicine,Hubei Shiyan 442012,China;
2.Department of Clinical Laboratory, Taihe Hospital Affiliated to Hubei Medicine University,Hubei Shiyan 442000,China
关键词:
甲基化;  耐药;  hMLH1;  MGMT;  5-氮杂-2'脱氧胞苷
分类号:
R734.2; R730.43
DOI:
10.3969/j.issn.1671-7414.2015.03.024
文献标志码:
A
摘要:
目的 观察5-氮杂-2'脱氧胞苷(5-Aza-Cde)对体外培养的顺铂(DDP)耐药株肺癌A549/DDP细胞hMLH1,MGMT基因启动子区DNA甲基化状态及其表达的影响,探讨肺癌细胞hMLH1和MGMT基因失活的机制及去甲基化制剂对hMLH1和MGMT基因表达的调控。方法 5-Aza-Cde处理体外1640培养的肺癌A549/DDP细胞,甲基化特异性PCR(MSP)法检测用药前后细胞hMLH1和MGMT基因的甲基化状态,RT-PCR法检测用药前后细胞hMLH1和MGMT mRNA的表达。结果 在对照组A549细胞当中hMLH1基因是非甲基化状态和高表达,而MGMT显示为低甲基化(部分甲基化)状态和高表达; 而在顺铂耐药株A549-DDP中,hMLH1和MGMT基因均显示高甲基化状态,mRNA表达下调。结论 hMLH1和MGMT基因甲基化修饰程度与mRNA的表达呈相关性改变,证明hMLH1,MGMT基因甲基化可能是A549对顺铂耐药的因素之一。
Abstract:
Objective To investigate the effects of 5-Aza-2'-deoxycytidine(5-Aza-Cde)on DNA methylation and expression of hMLH1 and MGMT gene in the human lung cancer cell line A549/DDP.Methods A549/DDP cells were cultured with RPMI 1 640 medium and were treated with 5 μmol/L DNA methyhransferase inhibitor 5-Aza-Cde.Methylation-specific polymerase chain reaetioll(MSP)was used to detect the promoter methylation state of the hMLH1 and MGMT gene.RT-PCR was used to detect the mRNA expression ofhMLH1 and MGMT before and after treatment with 5-Aza-Cde,respectively.Results Before treatment with 5-Aza-Cde,hMLH1 and MGMT expressions were absent,and promoter hypermethylation of the hMLH1 and MGMT gene were detected in A549 cells.After treatment with 5-Aza-Cde,the promoter regionof the hMLH1 and MGMT gene exhibited a demethylation state,and their mRNA expressions were increased.Conclusion Promoter hypermethyhtion is amajor mechanism of hMLH1 and MGMT gene silencing in human lung cancer cells,and can be reversed by the demethylating agent 5-Aza-Cde,which can regulate the expressions of the hMLH1 and MGMT gene.

参考文献/References:

[1] Bignamia M,Casorelli I,Karran P.Mismatch repair and responseto DNA-damaging antitumour therapies[J].Eur J Cancer,2003,39(15):2142-2149.
[2] Safar AM,Spencer H,Su X,et al.Promoter hypermethylation for molecularnodal staging in non-small cell lung cance[J].Arch Pathol Lab Med,2007,131(6):936-941.
[3] Lenz G,Hutter G,Hiddemann W,et al.Promoter methylation and expressionof DNA repair genes hMLH1 and MGMT in acute myeloid leukemia[J].Ann Hematol,2004,83(10):628-633.
[4] 陈英剑,胡成进,赵苗青.大鼠TGF-β1 mRNA表达水平的SYBR G reen Ⅰ荧光定量RT-PCR方法的建立[J].中国实验诊断学,2005,9(2):279-281.
Chen YJ,Hu CJ,Zhao MQ.Real-time reverse transcriptase-polymerase chain reaction for measurement of transforming growth factor beta 1 mRNA expression with SYBR green I[J].Chinese Journal of Laboratory Diagnosis,2005,9(2):279-281.
[5] Zhang H,Zhang SQ,Cui J,et al.Expression and promoter methylation status of mismatch repair gene hMLH 1 and hMSH2 in epithelial ovarian cancer[J].Aust N Z J Obstet Gynaecol,2008,48(5):505-509.
[6] Arnold CN,Goel A,Boland CR.Role of hMLHI promoter hypermethylation indrug resistance to 5-fluorouracil in colorectal cancer cell lines[J].Int J Cancer,2003,106(1):66-73.
[7] Li LS,Morales JC,Hwang A,et al.DNA mismatch repair-dependent activation of c-Abl/p73alpha/GADD45alpha-mediated apoptosis[J].J Biol Chem,2008,283(31):21394-21403.
[8] 孔云明,金永堂,薛绍礼,等.NSCLC患者血浆P16和MGMT基因甲基化检测及临床意义[J].肿瘤,2007,27(9):715-718.
Kong YM,Jing YT,Xue SL,et al.Detection and significance of promoter hypermethylation of P16 gene and MGMT gene in plasma from non-small cell lung cancer patients[J].Tumor,2007,27(9):715-718.
[9] Plimack ER,Stewart DJ,Issa JP,et al.Combining epigenetic and cytotoxic therapy in the treatment of solid tumors[J].Journal of Clinical Oncology,2007,25(29):4519-4521.
[10] 王 蒙,汤志刚,黄 强,等.5-氮杂-2-脱氧胞苷对人胰腺癌细胞株PANC-1侵袭和迁移能力的影响[J].肿瘤防治研究,2013,40(4):332-335.
Wang M,Tang ZG,Huang Q,et al.Effect of 5-Aza-CaR on invasion and migration of pancreatic carcinoma cell line PANG-I[J].Cancer Res Prey Treat,2013,40(4):332-335.

相似文献/References:

[1]陈 菲,曹小利,程 莉,等.尿培养产CTX-M大肠埃希菌的种系分型及耐药和毒力特点分析[J].现代检验医学杂志,2016,31(02):12.[doi:10.3969/j.issn.1671-7414.2016.02.004]
　CHEN Fei,CAO Xiao-li,CHENG Li,et al.Analysis on the Resistance and Virulence Characteristics and Phylogenetic Groups of CTX-M-Producing Escherichia Coli Isolated from Urine[J].Journal of Modern Laboratory Medicine,2016,31(03):12.[doi:10.3969/j.issn.1671-7414.2016.02.004]
[2]杨 健,王西娣,陈美玲,等.2013～2015年陕西地区结核分枝杆菌对利福平耐药性及rpoB基因突变的相关研究[J].现代检验医学杂志,2016,31(02):53.[doi:10.3969/j.issn.1671-7414.2016.02.016]
　YANG Jian,WANG Xi-di,CHEN Mei-ling,et al.Related Research on Resistance of Mycobacterium Tuberculosis to Rifampin and rpoB Gene Mutations in Shaanxi from 2013 to 2015[J].Journal of Modern Laboratory Medicine,2016,31(03):53.[doi:10.3969/j.issn.1671-7414.2016.02.016]
[3]谭皓妍,冯浩华,何艳屏.阴道白假丝酵母菌的耐药性与MDR1基因表达关系研究[J].现代检验医学杂志,2015,30(01):105.[doi:10.3969/j.issn.1671-7414.2015.01.029]
　TAN Hao-yan,FENG Hao-hua,HE Yan-ping.Research the Expression of MDR1 Genes in Vaginal Candida Albicans Resistance to Azole Agents[J].Journal of Modern Laboratory Medicine,2015,30(03):105.[doi:10.3969/j.issn.1671-7414.2015.01.029]
[4]肖 成,王玉明,胡 莹,等.社区相关性耐甲氧西林金黄色葡萄球菌USA300克隆的研究进展[J].现代检验医学杂志,2017,32(04):160.[doi:10.3969/j.issn.1671-7414.2017.04.046]
　XIAO Cheng,WANG Yu-ming,HU Ying,et al.Reserch Progress of USA300 Clone of Community-Associated Methicillin-Resistant Staphylococcus Aureus[J].Journal of Modern Laboratory Medicine,2017,32(03):160.[doi:10.3969/j.issn.1671-7414.2017.04.046]
[5]严海忠,王 娟,卢兰芬,等.糖尿病患者感染产ESBLs肠杆菌科细菌的耐药性及危险因素分析[J].现代检验医学杂志,2018,33(05):82.[doi:10.3969/j.issn.1671-7414.2018.05.022]
　YAN Hai-zhong,WANG Juan,LU Lan-fen,et al.Analysis of the Drug Resistance and Risk Factors of ESBLs-Producing Enterobacteriaceae Infection in Diabetics[J].Journal of Modern Laboratory Medicine,2018,33(03):82.[doi:10.3969/j.issn.1671-7414.2018.05.022]
[6]李晓丽,胡陇娟.子宫内膜癌组织中长链非编码RNA ZEB1-AS1的表达与临床特征及对化疗药物耐药性研究[J].现代检验医学杂志,2019,34(04):35.[doi:10.3969/j.issn.1671-7414.2019.04.009]
　LI Xiao-li,HU Long-juan.Research on Expression and Clinical Characteristics of Long-Chain Non-Coding RNA ZEB1-AS1 in Endometrial Carcinoma and Resistance to Chemotherapy Drugs[J].Journal of Modern Laboratory Medicine,2019,34(03):35.[doi:10.3969/j.issn.1671-7414.2019.04.009]
[7]曹 玲.长链非编码RNA ZEB1-AS1在儿童脑肿瘤中表达及对化疗药物敏感性影响的实验研究[J].现代检验医学杂志,2019,34(05):84.[doi:10.3969/j.issn.1671-7414.2019.05.021]
　CAO Ling.Experimental Study on the Expression Pattern of Long Non-Coding RNA ZEB1-AS1 and Its Effect on the Resistance of Chemotherapy on Childhood Brain Tumor[J].Journal of Modern Laboratory Medicine,2019,34(03):84.[doi:10.3969/j.issn.1671-7414.2019.05.021]
[8]王 娟,杨 柳,刘家云.肝癌患者血浆ctDNA SCNM1甲基化水平检测及与临床预后评估价值研究[J].现代检验医学杂志,2024,39(06):18.[doi:10.3969/j.issn.1671-7414.2024.06.003]
　WANG Juan,YANG Liu,LIU Jiayun.Plasma ctDNA SCNM1 Methylation Level and Its Clinical Prognostic Value in Hepatocellular Carcinoma Patients[J].Journal of Modern Laboratory Medicine,2024,39(03):18.[doi:10.3969/j.issn.1671-7414.2024.06.003]
[9]杨艳敏,郝秀君,赵志芳,等.骨髓增生异常综合征患者骨髓中ABAT mRNA表达和ABAT甲基化水平对预后预测价值研究[J].现代检验医学杂志,2024,39(06):84.[doi:10.3969/j.issn.1671-7414.2024.06.014]
　YANG Yanmin,HAO Xiujun,ZHAO Zhifang,et al.Prognostic Value of ABAT mRNA Expression and ABAT Methylation Level in Bone Marrow of Patients with Myelodysplastic Syndrome[J].Journal of Modern Laboratory Medicine,2024,39(03):84.[doi:10.3969/j.issn.1671-7414.2024.06.014]

备注/Memo

备注/Memo:
基金项目:湖北医药学院优秀中青年科技创新团队资助项目,十堰市科技局项目,项目编号:ZD2012015。
作者简介:王 虹(1964-),女,本科,副教授,研究方向:卫生事业管理及临床肿瘤治疗,E-mail:wanghong9763@163.com。
通讯作者:骆海军(1984-),男,硕士,主管检验师,研究方向:肿瘤与甲基化,E-mail:lhj403066210@163.com。
更新日期/Last Update: 2015-06-10