ABCB1和ABCG2在下咽癌FaDu细胞系多药耐药中的作用

[Role of ABCB1 and ABCG2 in the multidrug resistance of hypopharyngeal carcinoma FaDu cell line].

作者信息

MA Ju-ke, LU Su-mei, YU Liang, TIAN Jia-jun, LI Jian-feng, WANG Hai-bo, XU Wei

机构信息

Department of Otorhinolaryngology Head and Neck Surgery, Provincial Hospital Affiliated to Shandong University, Jinan 250021, China.

出版信息

Zhonghua Er Bi Yan Hou Tou Jing Wai Ke Za Zhi. 2012 Apr;47(4):305-10.

PMID:22800348

Abstract

OBJECTIVE

To investigate the expression of multidrug resistance gene ABCB1 and ABCG2 in FaDu cells (human hypopharyngeal carcinoma cell line) and the multidrug resistance (MDR) cell lines FaDu/T transformed from FaDu cells by taxol and underlying mechanisms of MDR.

METHODS

The multidrug resistance sensitivities of FaDu and FaDu/T to cisplatin (DDP), 5-fluorouracil (5-FU), doxorubicin (Dox), and vincristine (VCR) were examined by methyl-thiazolyl-tetrazolium (MTT) assay. The mRNA and protein expressions of multidrug resistance genes ABCB1 and ABCG2 were analysed with RT-PCR, Western blot and laser confocal microscopy. JNK signal proteins were detected through Western blot.

RESULTS

The multidrug resistance of FaDu/T cells to Taxol, DDP, 5-FU, ADM and VCR was more than that of FaDu cells. The expression of ABCB1 in FaDu/T cells was significantly higher than that in FaDu cells (t = 22.42, P < 0.05), but the expression of ABCG2 in FaDu/T cells was significantly lower than that in FaDu cells (t = 10.06, P < 0.05). JNK signal was inhibited in FaDu or FaDu/T cells and the inhibited JNK was reactivated by taxol or anisomycin (an activator for MAPK signal transduction pathways). Anisomycin down-regulated the expression of ABCB1 (F = 33.72, P < 0.05) and up-regulated the expression of ABCG2 (F = 220.16, P < 0.05) in FaDu/T cells, but not in FaDu/T cells pretreated by JNK inhibitor SP600125 (P > 0.05).

CONCLUSION

The overexpression of ABCB1 and the down-regulation of ABCG2 in FaDu/T cells were the main features of MDR in hypopharyngeal carcinomas, in which JNK signal transduction pathways could play an important role.

摘要

目的

研究多药耐药基因ABCB1和ABCG2在FaDu细胞（人下咽癌细胞系）及经紫杉醇诱导由FaDu细胞转化而来的多药耐药（MDR）细胞系FaDu/T中的表达情况及其多药耐药的潜在机制。

方法

采用甲基噻唑基四氮唑蓝（MTT）法检测FaDu和FaDu/T对顺铂（DDP）、5-氟尿嘧啶（5-FU）、阿霉素（Dox）和长春新碱（VCR）的多药耐药敏感性。运用逆转录-聚合酶链反应（RT-PCR）、蛋白质免疫印迹法（Western blot）和激光共聚焦显微镜分析多药耐药基因ABCB1和ABCG2的mRNA及蛋白表达。通过蛋白质免疫印迹法检测JNK信号蛋白。

结果

FaDu/T细胞对紫杉醇、DDP、5-FU、阿霉素和VCR的多药耐药性高于FaDu细胞。FaDu/T细胞中ABCB1的表达显著高于FaDu细胞（t = 22.42，P < 0.05），但FaDu/T细胞中ABCG2的表达显著低于FaDu细胞（t = 10.06，P < 0.05）。FaDu或FaDu/T细胞中的JNK信号受到抑制，而紫杉醇或茴香霉素（一种丝裂原活化蛋白激酶信号转导通路激活剂）可使受抑制的JNK重新激活。茴香霉素下调FaDu/T细胞中ABCB1的表达（F = 33.72，P < 0.05）并上调ABCG2的表达（F = 220.16，P < 0.05），但在经JNK抑制剂SP600125预处理的FaDu/T细胞中无此作用（P > 0.05）。