5-杂氮-2’-脱氧胞苷对前列腺癌PC-3细胞生物学行为的影响

Effect of 5-Aza-2＇-deoxycytidine on biological behaviors of prostate cancer PC-3 cell line

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、10．0μmol／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（FIT-PCR）方法检测E—cadherinmRNA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。

Objective To-detect the expression of E-cadherin and methylation status of CpG is- land 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 tLmol/L for72 h. The methyl- ation 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 （ RT- PCR） and Western blotting, respectively. The change of invasive ability was tested by using transwell cab- in. 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.