自噬基因Beclin1在卵巢癌SKOV3细胞中对PI3K/AKT信号通路的影响及其与紫杉醇耐药关系实验研究  

Effect of autophagy gene Beclin1 on PI3K/AKT signaling pathway in ovarian cancer SKOV3 cells and its correlation with paclitaxel resistance

作　　者：苏游 刘新月 刘巧梅 SU You;LIU Xinyue;LIU Qiaomei(Department of Gynecology,Guangxi International Zhuang Medicine Hospital,Nanning 530201,China)

机构地区：[1]广西国际壮医医院妇科,广西南宁530201

出　　处：《陕西医学杂志》2021年第2期143-147,共5页Shaanxi Medical Journal

基　　金：广西壮族自治区科技攻关项目(桂科攻12300016)。

摘　　要：目的:探究自噬基因Beclin1在卵巢癌SKOV3细胞中对PI3K/AKT信号通路的影响,并分析其与紫杉醇耐药的关系。方法:以人卵巢癌紫杉醇耐药细胞株SKOV3/TXA作为研究对象,构建自噬基因Beclin1真核表达载体pcDNA3.1/Beclin1,并通过试剂盒将其转染至SKOV3细胞中。采用MTT实验测定各组细胞增殖能力;流式细胞仪检测细胞凋亡及自噬情况,同时RT-PCR检测细胞中Beclin1 mRNA表达量,Western blot检测Beclin1、p-PI3K、PI3K、p-AKT和AKT蛋白水平。给予不同浓度紫杉醇处理,评估各组SKOV3/TXA细胞对紫杉醇耐药性的变化。结果:转染pcDNA3.1-Beclin1后,Beclin1-SKOV3/TXA组细胞Beclin1 mRNA和蛋白表达量明显升高(均P<0.05);细胞增殖能力较pcDNA3.1-SKOV3/TXA组和SKOV3/TXA组均显著降低(均P<0.05),凋亡率和自噬囊泡也明显增加(均P<0.05)。Western blot检测发现,Beclin1过表达后,SKOV3/TXA细胞p-PI3K/PI3K及p-AKT/AKT蛋白表达量比值显著低于pcDNA3.1-SKOV3/TXA组和SKOV3/TXA组(均P<0.05),磷酸化水平降低。2000 ng/ml和2500 ng/ml紫杉醇处理后,Beclin1-SKOV3/TXA组细胞凋亡率均高于其他组,差异均有统计学意义(均P<0.05)。结论:自噬基因Beclin1过表达可逆转SKOV3/TXA细胞的紫杉醇耐药性,其机制可能与抑制PI3K/Akt信号通路,诱导细胞自噬、凋亡有关。Objective:To investigate the effect of autophagy gene Beclin1 on the PI3K/AKT signaling pathway in ovarian cancer SKOV3 cells,and to analyze its relationship with paclitaxel resistance.Methods:SKOV3/TXA,a paclitaxel-resistant cell line of human ovarian cancer,was studied.The eukaryotic expression vector pcDNA3.1/Beclin1 of autophagy gene Beclin1 was constructed and transfected into SKOV3 cells by kit.The proliferation ability of cells in groups was determined by MTT assay.Apoptosis and autophagy were detected by flow cytometry.Beclin1 mRNA expression was detected by RT-PCR.Meanwhile,the protein levels of Beclin1,p-PI3K,PI3K,p-AKT and AKT were detected by Western blot.The paclitaxel resistance of each group was evaluated after intervention with different concentrations of paclitaxel.Results:After transfection with pcDNA3.1/Beclin1,the mRNA and protein levels of Beclin1 in Beclin1-SKOV3/TXA group were significantly increased(all P<0.05).In comparison with pcDNA3.1-SKOV3/TXA group and SKOV3/TXA group,the cell proliferation ability was significantly decreased,and the cell apoptosis and autophagy were significantly higher(all P<0.05).Western-blot results showed that after Beclin1 overexpression,the protein levels of p-PI3K/PI3K and p-AKT/AKT in Beclin1-SKOV3/TXA group were significantly lower than those in pcDNA3.1-SKOV3/TXA group and SKOV3/TXA group(all P<0.05),and the phosphorylation level was reduced.The cell apoptosis rate of Beclin1-SKOV3/TXA group were higher than that of other two groups after the intervention with 2000 ng/ml and 2500 ng/ml of paclitaxel(all P<0.05).Conclusion:Overexpression of the autophagy gene Beclin1 can reverse the paclitaxel resistance of SKOV3/TXA cells.The mechanism may be related to the inhibition of PI3K/AKT signaling pathway and the induction of cell autophagy and apoptosis.

关 键 词：自噬基因BECLIN1 SKOV3/TXA细胞 PI3K/AKT信号通路 紫杉醇耐药 卵巢癌 

分 类 号：R737.31[医药卫生—肿瘤]