[PI-3K/Akt信号通路抑制剂LY294002对卵巢癌细胞系A2780/Taxol多药耐药的逆转作用]

[Reversal effect of PI-3K/Akt pathway inhibitor LY294002 on multidrug resistance of ovarian cancer cell line A2780/Taxol].

作者信息

Shi Xiao-Yan, Cai Xiao-Jun, Lei Jian-Xiang, Cao Feng-Jun, Pan Dong-Feng, Chen Ping

机构信息

Department of Oncology, People's Hospital, Yunyang Medical College, Shiyan, Hubei, 442000, PR China.

出版信息

Ai Zheng. 2008 Apr;27(4):343-7.

PMID:18423117

Abstract

BACKGROUND & OBJECTIVE: Previous studies had showed that phosphatidylinositol 3 kinase (PI-3K) can suppress cell apoptosis. The inhibitor of PI-3K has been used to investigate the mechanisms of PI-3K-induced oncogenesis. This study was to investigate the reversal effect of LY294002, a PI-3K/Akt inhibitor, on paclitaxel-resistance of ovarian carcinoma cell line A2780/Taxol.

METHODS

A2780/Taxol cells were treated with LY294002. Cell apoptosis was analyzed by flow cytometry (FCM). The 50% inhibition concentration (IC50) of paclitaxel for A2780/Taxol cells was determined by MTT assay. The expression of multidrug resistance 1 (MDR1) mRNA was assessed by reverse transcription-polymerase chain reaction (RT-PCR). The expression of phosphated Akt and P-glycoprotein (P-gp) were detected by Western blot.

RESULTS

When treated for 24 h, the apoptosis rate of A2780/Taxol cells was significantly higher in 10 and 50 micromol/L LY294002 groups than in control group [(8.84+/-1.65)% and (20.78+/-2.47)% vs. (1.25+/-0.78)%, P<0.05], the IC50 of paclitaxel for A2780/Taxol cells was decreased significantly (P<0.01) with the highest reverse efficiency of (78.08+/-0.37)%. Moreover, the expression of MDR1 gene, and the phosphorylation of Akt and P-gp in A2780/Taxol cells were decreased.

CONCLUSIONS

The activation of PI-3K/Akt pathway plays an important role in paclitaxel-resistance of ovarian carcinoma cells. PI-3K/Akt inhibitor, LY294002 has a reversal effect on the paclitaxel-resistance of A2780/Taxol cells.

摘要

背景与目的

既往研究表明磷脂酰肌醇3激酶（PI-3K）可抑制细胞凋亡。PI-3K抑制剂已被用于研究PI-3K诱导肿瘤发生的机制。本研究旨在探讨PI-3K/Akt抑制剂LY294002对卵巢癌细胞系A2780/Taxol紫杉醇耐药的逆转作用。

方法

用LY294002处理A2780/Taxol细胞。采用流式细胞术（FCM）分析细胞凋亡情况。通过MTT法测定紫杉醇对A2780/Taxol细胞的50%抑制浓度（IC50）。采用逆转录-聚合酶链反应（RT-PCR）检测多药耐药1（MDR1）mRNA的表达。通过蛋白质免疫印迹法检测磷酸化Akt和P-糖蛋白（P-gp）的表达。

结果

处理24小时后，10和50μmol/L LY294002组A2780/Taxol细胞的凋亡率显著高于对照组[（8.84±1.65）%和（20.78±2.47）% vs.（1.25±0.78）%，P<0.05]，A2780/Taxol细胞对紫杉醇的IC50显著降低（P<0.01），最高逆转效率为（78.08±0.37）%。此外，A2780/Taxol细胞中MDR1基因的表达以及Akt和P-gp的磷酸化水平均降低。