5-杂氮-2'-脱氧胞苷对卵巢癌HO8910细胞WWOX基因甲基化状态及其生物学行为的影响  

Effects of 5-Aza-CdR on methylation of WWOX gene and biological behavior of ovarian cancer cell line HO8910

作　　者：闫洪超[1] 仝建业[1] 于楠[1] 韩秋峪[1] 魏敏[1] 陆晓媛[1] 

机构地区：[1]徐州医学院附属医院妇产科,徐州221002

出　　处：《现代妇产科进展》2012年第4期250-254,共5页Progress in Obstetrics and Gynecology

基　　金：徐州市科技发展基金计划项目(No:XF10C086)

摘　　要：目的:检测卵巢癌HO8910细胞株中WWOX基因甲基化状态,探讨5-杂氮-2'-脱氧胞苷(5-Aza-CdR)对WWOX基因甲基化状态及其对HO8910细胞株生物学行为的影响。方法:将HO8910细胞分为对照组与5-Aza-CdR处理组,采用甲基化特异性聚合酶链反应(MSP)方法检测两组细胞WWOX基因甲基化状态;蛋白印迹法检测两组细胞WWOX蛋白的表达。体外实验:分别用四甲基偶氮唑蓝(MTT)比色法、体外侵袭实验及流式细胞仪等分析5-Aza-CdR对HO8910细胞生物学活性的影响。体内实验:用处理前后的HO8910细胞株分别接种于BALB/c裸鼠腹腔,观察两组裸鼠的生存时间及肿瘤生长情况,并采用Western blot分别检测两组裸鼠肿瘤组织中WWOX蛋白的表达。结果:(1)WWOX基因在HO8910细胞株中呈甲基化状态,经5-Aza-CdR处理后呈去甲基化状态。(2)Western blot检测结果显示,5-Aza-CdR处理组WWOX蛋白表达水平高于对照组(0.71±0.023 vs 0.13±0.012,P<0.05)。(3)5-Aza-CdR处理组各时间点的细胞增殖能力较对照组下降(P<0.05)。(4)体外侵袭实验显示,5-Aza-CdR处理组穿膜细胞数较对照组少(92.2±4.7 vs 172.1±5.2,P<0.05)。(5)流式细胞检测显示5-Aza-CdR处理组中67.13%的细胞阻滞于G0/G1期,较对照组高(21.52%,P<0.05)。(6)裸鼠体内实验表明,5-Aza-CdR处理组裸鼠体内的致瘤能力明显低于对照组,且5-Aza-CdR处理组WWOX蛋白表达水平明显高于对照组(0.65±0.031 vs 0.25±0.047,P<0.05)。结论:5-Aza-CdR能逆转HO8910细胞WWOX基因甲基化状态,并抑制细胞增殖。Objective：To investigate WWOX gene methylation in ovarian cancer cell line HO8910 and explore the effects of 5-Aza-CdR on methylation state of WWOX gene and biological behavior of ovarian cancer cell line HO8910.Methods：HO8910 cells were divided into two groups：control group and 5-Aza-CdR treated group.The methylation state of WWOX gene was evaluated by methylation specific polymerase chain reaction（MSP）.Expression of WWOX protein was detected by Western blot.In vitro experiments：the biology effect of 5-Aza-CdR on HO8910 cell was analyzed through the methyl thiazolyl tetrazolium test,transwell assay and flow cytometry.In vivo experiments：the models were established in nude mice with two groups cells respectively.Survival time and the growth of tumor of all the nude mouse were observed.Expressions of WWOX protein in tumor tissue of the two groups were detected respectively by Western blot.Results：（1）Hypermethylation state of the WWOX gene was detected in HO8910 cell line,after treatment with 5-Aza-CdR,demethylation state was detected in HO8910 cell line.（2）Western blot showed that the expression of WWOX protein（0.71±0.023）was significantly higher than that of the control group（0.13±0.012）（P0.05）.（3）The growth rate at each time point of 5-Aza-CdR treated group cell was significantly lower than that of the control group.（4）In vitro invasion assay showed that invasion cell number in 5-Aza-CdR treated group（92.2±4.7）was significant lower than that of the control group（172.1±5.2,P0.05）.（5）Flow cytometry showed that 67.13% of cells was arrested G0/G1 stage in 5-Aza-CdR treated group,while control group was 21.52%（P0.05）.（6）In vivo test of nude mice showed that the tumorigenic ability of 5-Aza-CdR treated group was lower than that of the control group,and expression of WWOX protein（0.65±0.031）was significantly higher than that of the control group（0.25±0.047）（P0.05）.Conclusions：Methylation state of the WWOX gene in HO8910 cell line can be

关 键 词：卵巢肿瘤 WWOX基因 5-杂氮-2'-脱氧胞苷 小鼠 裸 

分 类 号：R737.31[医药卫生—肿瘤]