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目的:寻找与乳腺癌细胞株间药物敏感性差异相关的基因启动子CpG岛甲基化异常,为DNA甲基化异常作为临床化疗敏感性预测手段提供依据。方法:用MTT法测定3种乳腺癌细胞株(Bcap-37、T47D和ZR-75-30)对9种化疗药物(紫杉醇、紫杉特尔、长春瑞宾、5-氟尿嘧啶(5-FU)、双氟去氧胞苷、表阿霉素、丝裂霉素、依立替康和顺铂)的敏感性;同时用甲基化特异性PCR检测23个基因在3种乳腺癌细胞株中的甲基化状态。结果:受试的3种乳腺癌细胞株仅对5-FU的敏感性有明显差异。其中Bcap-37(IC50:317.386μg/mL)对5-FU相对耐药,而ZR-75-30(IC50:6.676μg/mL)和T47D(IC50:73.076μg/mL)对5-FU相对敏感。23个基因中,有6个基因:ABCC8、CHFR、BMP3B、LOX、PRSS21和RASSF1A在敏感和耐药细胞株中的甲基化状态有差异。其中ABCC8、CHFR和BMP3B在5-FU耐药细胞株中呈相对高甲基化状态,而LOX、PRSS21和RASSF1A则在对5-FU敏感的细胞株中呈相对高甲基化状态。结论:本研究所发现的6个基因的甲基化状态可能与乳腺癌细胞株对5-FU化疗敏感性相关,为下一步进行临床评估和机制探讨提供了依据。
OBJECTIVE: To find out the methylation abnormality of CpG island related to the difference of drug sensitivity between breast cancer cell lines and to provide evidence for the abnormality of DNA methylation as a means of predicting the sensitivity of clinical chemotherapy. Methods: Three chemotherapeutic agents (paclitaxel, paclitaxel, vinorelbine and 5-fluorouracil) were tested by MTT assay in three breast cancer cell lines (Bcap-37, T47D and ZR-75-30) , Difluorodeoxycytidine, epirubicin, mitomycin, irinotecan and cisplatin). Meanwhile, methylation-specific PCR was used to detect the methylation of 23 genes in three breast cancer cell lines State of affairs Results: The sensitivity of 3 breast cancer cell lines to 5-FU was significantly different. Among them, Bcap-37 (IC50: 317.386μg / mL) was relatively resistant to 5-FU while ZR-75-30 (IC50: 6.676μg / mL) and T47D (IC50: 73.076μg / mL) . Among the 23 genes, there are 6 genes: methylation status of ABCC8, CHFR, BMP3B, LOX, PRSS21 and RASSF1A in sensitive and resistant cell lines. Among them, ABCC8, CHFR and BMP3B were relatively hypermethylated in 5-FU resistant cell lines while LOX, PRSS21 and RASSF1A were relatively hypermethylated in 5-FU sensitive cell lines. CONCLUSION: The methylation status of the six genes found in this study may be related to the sensitivity of breast cancer cell lines to 5-FU chemotherapy, which provides a basis for further clinical evaluation and mechanism exploration.