Yao Lie, Gu Jichun, Mao Yishen, Zhang Xinju, Wang Xiaoyi, Jin Chen, Fu Deliang, Li Ji
Department of Pancreatic Surgery, Huashan Hospital, Fudan University, Shanghai 200040, P.R. China.
Department of Clinical Medicine, Fudan University, Shanghai 200082, P.R. China.
Oncol Lett. 2018 Apr;15(4):5602-5610. doi: 10.3892/ol.2018.8041. Epub 2018 Feb 13.
The main focus of the present study was to evaluate whether ABC transporter family promoter methylation predicted multidrug resistance in gemcitabine-resistant cancer cell lines (BxPC-3/Gem and PANC-1/Gem). Using low concentrations of gemcitabine, the cell lines acquired drug resistance with different initial gemcitabine concentrations. A novel technology, methylation-sensitive high-resolution melting, was used to monitor the dynamic changes of ABC transporter family promoter methylation, including ATP binding cassette subfamily B member 1 (ABCB1), ATP binding cassette subfamily C (ABCC) and ATP binding cassette subfamily G member 2 (ABCG2) mRNA expression. It was revealed that, with elevation of initial gemcitabine concentration, expression of ABCB1, ABCC and ABCG2 mRNA and corresponding downstream proteins was increased while promoter methylation was decreased. These discoveries indicate that promoter methylation of ABCB1, ABCC and ABCG2 may be a valuable indicator of drug-resistance characteristics in BxPC-3/Gem and PANC-1/Gem cells via quantitative and simultaneous detection. These results also implied that MDR in pancreatic cancer not only arises from gene mutation, but also originates from promoter methylation.
本研究的主要重点是评估ABC转运蛋白家族启动子甲基化是否可预测吉西他滨耐药癌细胞系（BxPC-3/Gem和PANC-1/Gem）中的多药耐药性。使用低浓度的吉西他滨，这些细胞系在不同的初始吉西他滨浓度下获得了耐药性。一种新技术，即甲基化敏感的高分辨率熔解技术，被用于监测ABC转运蛋白家族启动子甲基化的动态变化，包括ATP结合盒亚家族B成员1（ABCB1）、ATP结合盒亚家族C（ABCC）和ATP结合盒亚家族G成员2（ABCG2）的mRNA表达。结果显示，随着初始吉西他滨浓度的升高，ABCB1、ABCC和ABCG2的mRNA及相应下游蛋白的表达增加，而启动子甲基化减少。这些发现表明，通过定量和同时检测，ABCB1、ABCC和ABCG2的启动子甲基化可能是BxPC-3/Gem和PANC-1/Gem细胞耐药特征的一个有价值的指标。这些结果还暗示，胰腺癌中的多药耐药不仅源于基因突变，还源于启动子甲基化。
