摘要
背景与目的:磷脂酰肌醇(phosphatidylinositol 3 kinase,PI-3K)可抑制细胞凋亡,对PI-3K抑制剂的研究可以更好地了解PI-3K的促癌机制,为多种癌症如卵巢癌、乳腺癌等的基因治疗提供线索。本研究目的在于探讨PI-3K/Akt信号通路抑制剂LY294002对卵巢癌耐紫杉醇细胞株A2780/Taxol多药耐药的逆转作用。方法:用LY294002处理A2780/Taxol细胞,流式细胞术检测细胞凋亡,MTT法检测细胞对紫杉醇的药物敏感性,RT-PCR检测MDR1mRNA的表达,Western blot方法分析LY294002作用前后磷酸化Akt及P-gp蛋白的表达。结果:10和50μmol/L LY294002干预A2780/Taxol细胞24h后,A2780/Taxol细胞凋亡率分别为(8.84±1.65)%和(20.78±2.47)%,显著高于未干预细胞的凋亡率(1.25±0.78)%(P<0.05);A2780/Taxol细胞对紫杉醇的半数抑制浓度(IC50)显著降低(P<0.01),相对逆转效率最高可达(78.08±0.37)%;MDR-1mRNA、磷酸化Akt及P-gp蛋白均明显降低。结论:PI-3K/Akt信号通路的激活与卵巢癌细胞多药耐药的产生有关,PI-3K/Akt抑制剂LY294002可逆转卵巢癌细胞A2780/Taxol的多药耐药。
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 MTI- 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 pmol/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.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2008年第4期343-347,共5页
Chinese Journal of Cancer
