肿瘤研究与临床
腫瘤研究與臨床
종류연구여림상
CANCER RESEARCH AND CLINIC
2015年
3期
149-152
,共4页
余宗涛%张吉才%高琼%高波%胡春卉
餘宗濤%張吉纔%高瓊%高波%鬍春卉
여종도%장길재%고경%고파%호춘훼
肺肿瘤%hMLH1基因%甲基化%5-氮-2'脱氧胞苷%A549细胞%凋亡
肺腫瘤%hMLH1基因%甲基化%5-氮-2'脫氧胞苷%A549細胞%凋亡
폐종류%hMLH1기인%갑기화%5-담-2'탈양포감%A549세포%조망
Lung neoplasms%hMLH1 gene%Methylation%5-Aza-2'-deoxycytidine%A549 cells%Apoptosis
目的 探讨5-氮-2'脱氧胞苷(5-Aza-CdR)对耐顺铂(DDP)的人肺癌A549/DDP细胞凋亡及抑癌基因hMLH1表达的影响.方法 以浓度为0.5、5、50μmol/L的5-Aza-CdR处理A549/DDP细胞,常规培养,采用四甲基偶氮唑蓝(MTT)比色法观察细胞的生长活性,甲基化特异性聚合酶链反应(MSP)检测hMLH1基因甲基化状态,以实时荧光定量PCR法检测hMLH1 mRNA的表达,应用流式细胞术检测细胞凋亡率.结果 5-Aza-CdR能明显抑制肿瘤细胞的生长,随5-Aza-CdR浓度增加、培养时间延长,细胞生长抑制率升高(P＜0.05).细胞凋亡率与5-Aza-CdR剂量呈正相关(P＜ 0.001).5-Aza-CdR处理后hMLH1mRNA表达升高(P＜0.05).A549/DDP细胞hMLH1启动子甲基化阳性,其mRNA为阴性,应用5-Aza-CdR干预培养后,mRNA为阳性.结论 5-Aza-CdR能使hMLH1基因去甲基化,促进细胞凋亡,增强抑癌功能.
目的 探討5-氮-2'脫氧胞苷(5-Aza-CdR)對耐順鉑(DDP)的人肺癌A549/DDP細胞凋亡及抑癌基因hMLH1錶達的影響.方法 以濃度為0.5、5、50μmol/L的5-Aza-CdR處理A549/DDP細胞,常規培養,採用四甲基偶氮唑藍(MTT)比色法觀察細胞的生長活性,甲基化特異性聚閤酶鏈反應(MSP)檢測hMLH1基因甲基化狀態,以實時熒光定量PCR法檢測hMLH1 mRNA的錶達,應用流式細胞術檢測細胞凋亡率.結果 5-Aza-CdR能明顯抑製腫瘤細胞的生長,隨5-Aza-CdR濃度增加、培養時間延長,細胞生長抑製率升高(P＜0.05).細胞凋亡率與5-Aza-CdR劑量呈正相關(P＜ 0.001).5-Aza-CdR處理後hMLH1mRNA錶達升高(P＜0.05).A549/DDP細胞hMLH1啟動子甲基化暘性,其mRNA為陰性,應用5-Aza-CdR榦預培養後,mRNA為暘性.結論 5-Aza-CdR能使hMLH1基因去甲基化,促進細胞凋亡,增彊抑癌功能.
목적 탐토5-담-2'탈양포감(5-Aza-CdR)대내순박(DDP)적인폐암A549/DDP세포조망급억암기인hMLH1표체적영향.방법 이농도위0.5、5、50μmol/L적5-Aza-CdR처리A549/DDP세포,상규배양,채용사갑기우담서람(MTT)비색법관찰세포적생장활성,갑기화특이성취합매련반응(MSP)검측hMLH1기인갑기화상태,이실시형광정량PCR법검측hMLH1 mRNA적표체,응용류식세포술검측세포조망솔.결과 5-Aza-CdR능명현억제종류세포적생장,수5-Aza-CdR농도증가、배양시간연장,세포생장억제솔승고(P＜0.05).세포조망솔여5-Aza-CdR제량정정상관(P＜ 0.001).5-Aza-CdR처리후hMLH1mRNA표체승고(P＜0.05).A549/DDP세포hMLH1계동자갑기화양성,기mRNA위음성,응용5-Aza-CdR간예배양후,mRNA위양성.결론 5-Aza-CdR능사hMLH1기인거갑기화,촉진세포조망,증강억암공능.
Objective To investigate the effects of 5-Aza-2'-deoxycytidine (5-Aza-CdR) on the apoptosis of A549/DDP cells and the expression of hMLH1 gene.Methods A549/DDP cells were treated with 5-Aza-CdR at 0.5,5,50 μmol/L.The growth curve of A549/DDP cells was investigated by MTT assay.The methylation status of hMLH1 gene was detected by methylation specific PCR (MSP).The expression of hMLH1 mRNA was evaluated by FQ-PCR.The apoptosis rate of A549/DDP cells was analyzed by flow cytometry.Results A549/DDP cells treated with 5-Aza-CdR showed a slow growth in comparison with the control cells,and the growth rates were decreased with the increasing of 5-Aza-CdR concentration.The apoptosis rate after treatment was higher than that before treatment in A549/DDP cells (P ＜ 0.05),and had a positive correlation with 5-Aza-CdR dose (P ＜ 0.001).hMLH1 mRNA expression level was increased in a 5-Aza-CdR concentration dependent manner (P ＜ 0.05).hMLH1 promoter in A549/DDP cells was methylated and hMLH1 mRNA was negatively expressed before treatment,but the mRNA was positively expressed after treatment with 5-Aza-CdR.Conclusions 5-Aza-2'-CdR can induce apoptosis of A549/DDP cells by inducing demethylation of hMLH1 promoter and thereby enhancing hMLH1 gene expression and its tumor suppressor function.
