中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
10期
1769-1771
,共3页
前列腺癌%E-cadherin%甲基化%5-杂氮-2′-脱氧胞苷
前列腺癌%E-cadherin%甲基化%5-雜氮-2′-脫氧胞苷
전렬선암%E-cadherin%갑기화%5-잡담-2′-탈양포감
Prostate cancer%E-cadherin%Methylation%5-Aza-2′-deoxycytidine
目的 检测E-cadherin基因在激素非依赖性前列腺癌(HIPC)细胞上的表达及启动子CpG岛甲基化,探讨甲基化抑制剂5-杂氮-2′-脱氧胞苷(5-Aza-CdR)对HIPC细胞的影响及意义.方法 2.5、5.0、10.0 μmol/L的5-Aza-CdR处理PC-3细胞72h后,甲基化特异性聚合酶链反应(MSP)方法检测CpG岛甲基化改变,逆转录—聚合酶链反应(RT-PCR)方法检测E-cadherin mRNA变化,Westernblot方法检测E-cadherin蛋白变化,Transwell小室检测细胞侵袭性改变.结果 HIPC的E-cadherin启动子CpG岛甲基化呈阳性,基因表达缺失,细胞侵袭性明显.经5-Aza-CdR作用之后,CpG岛甲基化阳性明显减弱(P＜0.05),E-cadherin基因恢复表达(P＜0.05),PC-3细胞的侵袭性下降约59.68％,且与药物的浓度呈正相关.结论 5-Aza-CdR可逆转PC-3细胞E-cadherin启动子CpG岛的异常甲基化,诱导mRNA转录和蛋白的表达,并降低癌细胞的侵袭性.
目的 檢測E-cadherin基因在激素非依賴性前列腺癌(HIPC)細胞上的錶達及啟動子CpG島甲基化,探討甲基化抑製劑5-雜氮-2′-脫氧胞苷(5-Aza-CdR)對HIPC細胞的影響及意義.方法 2.5、5.0、10.0 μmol/L的5-Aza-CdR處理PC-3細胞72h後,甲基化特異性聚閤酶鏈反應(MSP)方法檢測CpG島甲基化改變,逆轉錄—聚閤酶鏈反應(RT-PCR)方法檢測E-cadherin mRNA變化,Westernblot方法檢測E-cadherin蛋白變化,Transwell小室檢測細胞侵襲性改變.結果 HIPC的E-cadherin啟動子CpG島甲基化呈暘性,基因錶達缺失,細胞侵襲性明顯.經5-Aza-CdR作用之後,CpG島甲基化暘性明顯減弱(P＜0.05),E-cadherin基因恢複錶達(P＜0.05),PC-3細胞的侵襲性下降約59.68％,且與藥物的濃度呈正相關.結論 5-Aza-CdR可逆轉PC-3細胞E-cadherin啟動子CpG島的異常甲基化,誘導mRNA轉錄和蛋白的錶達,併降低癌細胞的侵襲性.
목적 검측E-cadherin기인재격소비의뢰성전렬선암(HIPC)세포상적표체급계동자CpG도갑기화,탐토갑기화억제제5-잡담-2′-탈양포감(5-Aza-CdR)대HIPC세포적영향급의의.방법 2.5、5.0、10.0 μmol/L적5-Aza-CdR처리PC-3세포72h후,갑기화특이성취합매련반응(MSP)방법검측CpG도갑기화개변,역전록—취합매련반응(RT-PCR)방법검측E-cadherin mRNA변화,Westernblot방법검측E-cadherin단백변화,Transwell소실검측세포침습성개변.결과 HIPC적E-cadherin계동자CpG도갑기화정양성,기인표체결실,세포침습성명현.경5-Aza-CdR작용지후,CpG도갑기화양성명현감약(P＜0.05),E-cadherin기인회복표체(P＜0.05),PC-3세포적침습성하강약59.68％,차여약물적농도정정상관.결론 5-Aza-CdR가역전PC-3세포E-cadherin계동자CpG도적이상갑기화,유도mRNA전록화단백적표체,병강저암세포적침습성.
Objective To detect the expression of E-cadherin and methylation status of CpG island in hormone independent prostate cancer (HIPC) cells,and to explore the influence and significance of DNA methyltransferase inhibitor 5-Aza-2′-deoxycytidine (5-Aza-CdR) on HIPC cells.Methods The PC-3 cells were treated with 5-Aza-CdR at concentrations of 2.5,5.0,10.0 μmol/L for72 h.The methylation of CpG island was measured by using methylation-spectific PCR (MSP) methods.The mRNA and protein expression of E-cadherin was detected by using reverse transcription polymerase chain reaction ( RTPCR) and Western blotting,respectively.The change of invasive ability was tested by using transwell cabin.Results E-cadherin gene methylation was positive,the expression of E-cadherin mRNA and protein was undetectable,and the invasive ability was strong in PC-3 cells.After treatment wtih 5-Aza-CdR,E-cadherin methylation was obviously weakened ( P ＜ 0.05 ),the expression of E-cadherin was refreshed (P ＜0.05),and the invasive ability of PC-3 cells was decreased by about 59.68％ (P ＜0.01 ) in a concentration-dependent manner.Conclusion 5-Aza-CdR can reverse the methylation of E-cadherin CpG island,promote the expression of E-cadherin mRNA and protein,and reduce the invasive ability of cancer cells.
