ABCC8、CHFR、BMP3B、LOX、PRSS21和RASSF1A基因启动子的甲基化状态可能与3种乳腺癌细胞株对5-FU的敏感性相关

The methylation status of ABCC8,CHFR,BMP3B,LOX,PRSS21,and RASSF1A promoters correlate with the sensitivity of three breast cancer cell strains to 5-FU

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 identify the abnormal chemosensitivity-associated DNA methylation pattern of CpG islands in the pro- moter region of breast cancer cell lines and provide the evidence for using abnormal DNA methylation in predicting the chemosensitivity in clinic. Methods：The MTT/IC50 assay was used to determine the chemosensifivity of the three breast cancer cell lines （ Beap-37, T47D, and ZR-75-30） to each of the nine anticancer drugs （paclitaxel, docetaxel, vinorelbine, 5-fluorouracil, gemeitabine, epirubiein, mitomycin, ironoteean, and cisplatin）. DNA methylation profile of CpG islands in the promoter region of 23 genes of the three breast cancer cell lines was tested by methylafion specific PCR. Results： The sensitivity of the three breast cancer cell lines to 5-FU showed a significant difference. Bcap-37 cells were relatively tolerant to 5-FU （IC50 = 317. 386 μg/mL）. ZR-75-30 and T47D were relatively sensitive to 5-FU （ICso = 6. 676 and 73. 076μg/mL, respectively）. Out of the 23 genes, the methylation of 6 genes, ABCC8, CHFR, BMP3B, LOX, PRSS21, and RASSF1A, exhibited different patterns between the 5-FU-resistant and sensitive cell lines. The ABCC8, CHFR and BMP3B genes were hypermethylated in the 5-FU-resistant cell line, but LOX, PRSS21, and RASSF1A genes were hypermethylated in the 5-FU-sensitive cell line. Conclusion：There is a promising correlation of the methylation state of 6 genes with the chemosensitivity of the breast cancer cell lines to 5-FU. This study provided the evidence for further clinical evaluation and mechanism exploration.