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目的探讨Wnt通路抑制因子甲基化与卵巢癌耐药的相关性。方法2008年在四川大学华西第二医院采用甲基化特异性PCR(MSP)、RT-PCR检测卵巢癌细胞株SKOV3、OVCAR-8(紫杉醇耐药),A2780及A2780/D(顺铂耐药)中,Wnt通路抑制因子SFRP-1、2、4、5,WIF-1、DKK-3的甲基化状态、mRNA水平;四甲基偶氮唑蓝(MTT)法比较DNA甲基转移酶抑制剂5-氮-2-脱氧胞苷(5-Aza-CdR)处理后,OVCAR-8细胞耐药逆转的情况,并检测WIF-1基因甲基化状态、mRNA表达水平的改变。结果WIF-1基因在OVCAR-8细胞中为高甲基化表达抑制,5-Aza-CdR可使其去甲基化后重新表达,并部分逆转OVCAR-8细胞的紫杉醇耐药。结论WIF-1基因的高甲基化状态与卵巢癌耐药细胞株OVCAR-8的紫杉醇耐药相关,去甲基化后可部分逆转耐药,这为逆转卵巢癌耐药提供了新的研究方向。
Objective To investigate the correlation between Wnt pathway inhibitor methylation and drug resistance in ovarian cancer. Methods Methylation-specific PCR (MSP) and ovarian cancer cell line SKOV3, OVCAR-8 (paclitaxel resistance), A2780 and A2780 / D were detected by RT-PCR in the Second West China Hospital of Sichuan University in 2008. ), The methylation status and mRNA levels of Wnt pathway inhibitor SFRP-1, 2, 4, 5, WIF-1 and DKK-3 were detected by MTT assay. Compared with DNA methyltransferase After treatment with 5-azacytidine (5-Aza-CdR), the resistance of OVCAR-8 cells was reversed and the methylation status and mRNA expression of WIF-1 gene were detected. Results The WIF-1 gene was highly methylated in OVCAR-8 cells. 5-Aza-CdR could be demethylated and then re-expressed, and partially reversed paclitaxel resistance in OVCAR-8 cells. Conclusion The hypermethylation status of WIF-1 gene is related to the drug resistance of ovarian cancer cell line OVCAR-8. Demethylation can partially reverse the drug resistance, which provides a new research direction for reversing the drug resistance of ovarian cancer.