摘要
目的探讨survivin反义寡核苷酸(ASODN)对乳腺癌细胞系MCF-7的诱导凋亡、抑制增殖和对长春瑞滨的增敏作用。方法采用脂质体介导survivin ASODN转染乳腺癌细胞系MCF-7,半定量RT-PCR方法检测survivin基因mRNA表达,Western blot方法检测survivin基因蛋白表达,AnnexinⅤ/PI双染法通过流式细胞仪检测细胞凋亡率变化,PI染色通过流式细胞仪检测细胞周期时相变化,MTT比色法检测转染细胞的生长抑制情况及对长春瑞滨的敏感性,电镜观察反义转染对乳腺癌细胞系MCF-7的凋亡诱导作用。结果survivin反义复合物下调MCF-7细胞的survivin mRNA和蛋白表达,并呈剂量依赖性。细胞周期被阻滞于G2/M期,细胞凋亡率明显升高(P<0.05),反义组细胞生长抑制率明显上升(P<0.05)。透射电镜下转染组细胞呈凋亡晚期变化。600 nmol/L反义转染组长春瑞滨的IC50值为(3.7±0.42)μg/mL,正常组为(7.1±0.68)μg/mL,差异有统计学意义(P<0.05)。结论survivin反义寡核酸可有效下调MCF-7的survivin mRNA及蛋白表达水平,抑制增殖,诱导凋亡,并增强其对化疗药物长春瑞滨的敏感性。
Objective To explore the effects of survivin antisense oligonucleotide(ASODN) on apoptosis,growth and sensitivity to vinorelbine in the human breast cancer cell MCF-7.Methods Survivin-ASODN was transfected into the MCF-7 cells mediated by the Lip reagent.Expression of survivin mRNA was determined by RT-PCR and of survivin protein by Western blot.Apoptosis rate and cell cycle changes in antisense oligonucleotide(ASODN) transfected cells were detected by FCM.Cell proliferation and chemo-sensitivity to vinorel...
出处
《山东大学学报（医学版）》
CAS
北大核心
2009年第1期45-49,共5页
Journal of Shandong University:Health Sciences
