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目的 :了解 E-钙粘连素 (E- cad)基因启动子 Cp G位点甲基化与前列腺癌细胞 E- cad基因的失活的关系 ,并探讨 E- cad基因异常甲基化机制。方法 :采用硫酸氢钠基因组测序法检测良性前列腺上皮和前列腺癌细胞株的 E- cad基因甲基化状态 ,并采用 RT- PCR检测各细胞株 E- cad和 DNA甲基转移酶 (Dnmtl)的 m RNA表达。结果 :有 33% (2 /6)的前列腺癌细胞株发生甲基化现象 ,相应的细胞株中 E- cad m RNA水平降低。用去甲基化剂偶氮胞嘧啶 (Aza C)处理后 ,能恢复 E-cad阴性的前列腺癌细胞株中 E- cad m RNA水平。前列腺癌细胞株比正常上皮细胞株有较高水平的 Dnmtl表达。结论 :前列腺癌细胞 E- cad基因的高甲基化状态是引起该基因失活的原因 ,基因的异常甲基化可能与 Dnmtl有关。
OBJECTIVE: To investigate the relationship between CpG methylation of promoter of E-cadherin gene and inactivation of E-cad gene in prostate cancer cells and explore the mechanism of abnormal methylation of E-cadherin gene. Methods: The methylation status of E-cad gene in benign prostate epithelial and prostate cancer cell lines was detected by sodium bisulfate sequencing and the expression of E- cad and DNA methyltransferase (Dnmtl) was detected by RT-PCR m RNA expression. RESULTS: Thirty-three percent (2/6) of prostate cancer cell lines were methylated, and the corresponding cell lines had a decreased level of E-cad m RNA. Treatment with the demethylating agent Azacitidine (Aza C) restored E-cad m RNA levels in E-cad-negative prostate cancer cell lines. Prostate cancer cell lines have higher levels of Dnmtl expression than normal epithelial cell lines. Conclusion: The hypermethylation status of E-cadherin in prostate cancer cells is the cause of inactivation of this gene. The abnormal methylation of the gene may be related to Dnmtl.