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survivin反义寡核苷酸转染对人乳腺癌细胞系
MCF7生长及对长春瑞滨的增敏作用

扈煜,黎莉,李丽珍
  

  1. (山东大学齐鲁医院肿瘤中心, 济南 250012)
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-01-16 发布日期:2009-01-16
  • 通讯作者: 黎莉

Effect of survivin antisense oligonucleotide antisense transfection on growth and sensitivity to vinorelbine in the human breast cancer cell MCF7

HU Yu, LI Li, LI Lizhen
  

  1. (Cancer Center, Qilu Hospital of Shandong University, Jinan 250012, China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-01-16 Published:2009-01-16
  • Contact: LI Li

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。

关键词: 乳腺肿瘤, survivin 寡核苷酸类,反义, 增殖, 凋亡, 长春瑞滨

Abstract:

To explore the effects of survivin antisense oligonucleotide(ASODN) on apoptosis, growth and sensitivity to vinorelbine in the human breast cancer cell MCF7. MethodsSurvivin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 vinorelbine in MCF7 were determined by the MTT assay. The effect of survivin antisense oligonucleotide(ASODN) in inducing apoptosis of MCF7 cells was observed under an electron microscope. ResultsSurvivin ASODNLip compound efficiently downregulated the mRNA and protein expressions of survivin in MCF7 cell lines in a concentrationdependent manner. The cell cycle was arrested at the G2/M phase, the cell apoptosis rate was increased obviously(P<0.05), and growth inhibition was evidently increased in antisense oligonucleotide(ASODN) transfected cells(P<0.05). Terminal apoptosis changes were observed under an electron microscope after antisense oligonucleotide(ASODN) transfection. The 50% inhibitory concentration(IC50) of vinorelbine was significantly lower for 600?nmol/L antisense oligonucleotide(ASODN) transfected MCF7 cells than for control cells [(3.7±0.42)μg/mL ConclusionSurvivin

Key words: Breast neoplasms, Survivin oligoribonucleotides,antisense, Cell proliferation, Apoptosis, Vinorelbine

中图分类号: 

  • R737.9
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