榄香烯对人肺癌A549细胞系耐顺铂细胞株多药耐药的改善作用及机制研究

Study on the Improvement Effect and Mechanism of Elemene on Multidrug Resistance of Eisplatin-resistant Human Lung Cancer A549 Cell Line

[目的]探究榄香烯(elemene,ELE)对顺铂(cisplatin,DDP)耐药人肺癌A549/DDP细胞系多药耐药的改善作用及其作用机制。[方法]将A549/DDP细胞分为A549/DDP组和ELE组,以四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)检测24、48、72h时不同浓度ELE对A549/DDP细胞增殖能力的影响。采用DDP及20、40μg·mL^-1 ELE处理A549/DDP细胞,MTT法检测ELE对细胞化疗敏感性的影响,Transwell侵袭实验检测细胞侵袭能力变化,逆转录聚合酶链式反应(reverse transcription-polymerase chain reaction,RT-PCR)检测B细胞淋巴瘤-2(B-cell lymphoma-2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2-associated X,Bax)mRNA表达水平,Western blot检测多药耐药基因1(multidrug resistance gene1,MDR1)、肺耐药相关蛋白(lung resistancerelated protein,LRP)及磷脂酰肌醇-3-激酶(phosphatidy linositol-3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)信号通路相关蛋白表达水平。[结果]MTT结果显示,ELE呈时间和剂量依赖性抑制A549/DDP细胞的增殖能力(均P<0.05)。与单独使用DDP比较,20和40μg·mL^-1 ELE提高A549/DDP细胞对DDP的敏感性,降低DDP对A549/DDP细胞的半数抑制浓度(50%inlibiting concentration,IC50),抑制细胞的增殖能力(均P<0.05)和侵袭能力(P<0.05,P<0.01),增加细胞Bax mRNA表达水平,减少Bcl-2 mRNA表达水平(均P<0.05),使A549/DDP细胞MDR1、LRP、磷酸化磷脂酰肌醇-3-激酶(phosphorylated phosphatidylinositol-3-kinase,p-PI3K)和磷酸化蛋白激酶B(phosphorylated protein kinase B,p-AKT)蛋白表达水平显著降低(均P<0.05)。[结论]ELE可增加A549/DDP细胞对DDP的敏感性,改善多药耐药现象,其作用机制可能与抑制PI3K/AKT信号通路的活化有关。

[Objective]To investigate the improvement effect and mechanism of elemene(ELE)on the multidrug resistance of cisplatin(DDP)-resistant human lung cancer A549/DDP cell line.[Methods]A549/DDP cells were divided into A549/DDP group and ELE group.Methyl thiazolyl tetrazolium(MTT)method was used to detect the effect of ELE on proliferation of A549/DDP cells at 24,48 and 72 h.MTT method was used to detect the effects of ELE on the sensitivity of A549/DDP cells to chemotherapy drugs.Transwell invasion test was used to detect the changes in cell invasion ability.Reverse transcription-polymerase chain reaction(RT-PCR)methods were used to detect B-cell lymphoma-2(Bcl-2)and Bcl-2-associated X(Bax)mRNA expression levels in cells.Western blot was used to detect multidrug resistance gene1(MDR1),lung resistance-related protein(LRP)and phosphatidylinositol-3-kinase(PI3K)/protein kinase B(AKT)signaling pathway-related protein expression levels.[Results]MTT test results showed that ELE inhibited the proliferation of A549/DDP cells in a time and dose-dependent manner(all P<0.05).Compared with DDP alone,20 and 40μg·mL^-1 ELE increased the sensitivity of A549/DDP cells to DDP,decreased the 50%inhibiting concentration(IC50)of DDP to A549/DDP cells,and inhibited cell proliferation(all P<0.05)and invasion ability(P<0.05,P<0.01),increased the expression level of Bax mRNA,decreased the expression level of Bcl-2 mRNA(all P<0.05),and significantly reduced the expression levels of MDR1,LRP,phosphorylated phosphatidylinositol-3-kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT)protein in A549/DDP cells(all P<0.05).[Conclusion]ELE can increase the sensitivity of A549/DDP cells to DDP and improve multidrug resistance.Its mechanism of action may be related to inhibiting the activation of PI3K/AKT signaling pathway.