基础医学与临床 ›› 2009, Vol. 29 ›› Issue (5): 453-458.

• 研究论文 • 上一篇    下一篇

NSPc1是维持HeLa细胞正常增殖的必要因子

胡光宇 吴旭东 彭小忠 龚燕华   

  1. 中国医学科学院基础医学研究所 北京协和医学院基础学院 医学分子生物学国家重点实验室 中国医学科学院基础医学研究所 北京协和医学院 医学分子生物学国家重点实验室 中国医学科学院 基础医学研究所 北京协和医学院 基础学院 医学分子生物学国家重点实验室 中国医学科学院基础医学研究所 北京协和医学院基础学院 医学分子生物学国家重点实验室
  • 收稿日期:2009-02-26 修回日期:2009-03-02 出版日期:2009-05-25 发布日期:2009-05-25
  • 通讯作者: 龚燕华

NSPc1 is a Necessary Factor for HeLa Cells' Proliferation

Guang-yu HU, Xu-dong WU, Xiao-zhong PENG, Yan-hua GONG   

  1. National Laboratory of Medical Molecular Biology, IBMS, CAMS & PUMC National Laboratory of Medical Molecular Biology, IBMS, CAMS&PUMC National Lab of Medical Molecular Biology, Institute of Basic Medical Science& School of Basic Medicine, CAMS & PUMC National Laboratory of Medicine Molecular Biology, IBMS, CAMS,School of Basic Medicine PUMC
  • Received:2009-02-26 Revised:2009-03-02 Online:2009-05-25 Published:2009-05-25
  • Contact: Yan-hua GONG

摘要: 目的 研究多梳蛋白家族成员NSPc1对HeLa细胞增殖的影响。方法 运用生物信息学设计并合成敲低NSPc1表达的siRNA序列,用半定量RT-PCR、Real-time PCR及Western blot等检测siRNA序列在HeLa细胞中敲低NSPc1的有效性;将相应有效序列构建到pSUPER载体中,观察其瞬时转染对HeLa细胞BrdU掺入的影响;用G418筛选并建立稳定敲低NSPc1的HeLa细胞系,观察敲低NSPc1对HeLa细胞体外增殖能力的影响。结果 (1)设计的siRNA序列可显著敲低NSPc1的表达;(2)瞬时敲低NSPc1阻碍了HeLa细胞BrdU的掺入;(3)成功构建稳定敲低NSPc1的HeLa细胞系,其增殖速度较对照组明显变慢;结论 NSPc1的正常表达是维持HeLa细胞正常增殖能力所必需的,稳定敲低NSPc1的HeLa细胞群可作为进一步研究NSPc1相关信号通路的有效模型。

关键词: Polycomb, NSPc1, HeLa细胞, 稳定细胞系, 增殖

Abstract: Objective To study the effect of PcG member NSPc1 on the proliferation of HeLa cells. Methods Using bioinfomatic analysis to design the siRNA sequence for knockdown NSPc1. Detecting the expression level of NSPc1 in HeLa cell lines using semi-quantitative RT-PCR, Real-time PCR and Western blot after transfection of the designed siRNA. Transient transfecting pSUPER-NSPc1 into Hela cells and performing BrdU incorporation assay. Establishing NSPc1 stably knockdown cell lines, comparing proliferation abilities with the control cells. Results 1) The designed siRNA could efficiently knockdown the expression of NSPc1; 2) Transient knockdown NSPc1 could repress BrdU incorporation; 3) The established NSPc1-knockdown cell lines had a significantly low proliferation rate than control cells. Conclusion The expression of NSPc1 is necessary for the normal proliferation of HeLa cells.The NSPc1 stably knockdown cell pools is a useful model for further study the pathway related to NSPc1.

Key words: Polycomb, NSPc1, HeLa cell, Stable cell line, Proliferation

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