PRACTICAL ONCOLOGY JOURNAL ›› 2010, Vol. 24 ›› Issue (4): 368-372.doi: 10.3969/j.issn.1002-3070.2010.04.020

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Influence of survivin gene expression inhibition with siRNA on the cell cycle of HeLa cell line

XU Xiaoying, LI Xiurong, LI Na, LEI Hongwei, SU Xu, LI Guoquan   

  1. Department of Radiotherapy,The Second Affiliated Hospital of Da Lian Medical University,Da lian 116027
  • Received:2010-04-27 Online:2010-08-28 Published:2015-01-24

Abstract: Objective To design siRNA targeting to survivin and construct an eukaryocytic expression vector pSUPER/survivin,then transfected it into cervical cancer cell line HeLa and evaluated the effects of siRNA against survivin gene on cell cycle of cervical cancer cell line HeLa irradiated with γ-rays.Methods The primer was synthesized,siRNA targeting to survivin was obtained,linked with the vector pSUPER and sequenced.The correct pSUPER/survivin was transfected into HeLa cells and the expression of survivin gene in Hela cells was detected by RT-PCR and Flow Cytometry(FCM).In the present study,the effects of survivin gene transfection combined with irradiation on cell cycle distribution were analysed by FCM.Result Recombinant eukaryocytic expression vector pSUPER/survivin was successfully constructed.FCM results indicated that the expression of survivin gene is significantly suppressed in pSUPER/survivin transfected cells compared with non-transfected cells or empty vector pSUPER transfected cells.After γ-ray irradiation,HeLa cells transfected with pSUPER/survivin induced less proportion of S phase and G2/M arrest.Conclusion Recombinant eukaryocytic expression vector pSUPER/survivin was successfully constructed.siRNA against survivin enhanced radiosensitivity through execution of apoptosis,induction less proportion of S phase and G2/M arrest,it may be a potential target for improving radiosensitivity

Key words: siRNA, survivin gene, γ-ray, HeLa cell, Cell cycle

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