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抗凋亡基因survivin促进细胞转化的作用机制 预览 被引量:61

Anti-apoptosis gene survivin promotes cell growth and transformation
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摘要 目的:探讨抗凋亡基因survivin在细胞转化中的作用机制。方法:利用RT-PCR获得survivin编码区序列,双酶切后,将其定向克隆至原核和真核表达载体,生长曲线观察高达到survivin对细胞增殖的影响;软琼脂集落形成实验观察细胞的接触抑制生长能力;WesternBlot检测survivin高表达引起的蛋白质分子改变。结果:通过分子克隆,构建survivin原核和真核表达载体,并可在大肠杆菌和真核细胞中表达。生长曲线表明,转染survivin的293细胞的增殖速度略快于对照细胞,尤其在细胞生长至高密度时;survivin可以明显提高293细胞的软琼脂集落形成能力;survivin的高表达可以引起癌基因cylinD1的蛋白表达水平增高。结论:抗凋亡基因survivin具有促进细胞转化的作用,这种作用可能依赖于cyclinD1和c-myc的功能。 Objective To investigate the role and molecular mechanism of suvivin, an anti apoptosis gene, in cell growth and transformation. Methods Coding sequence of survuvin was amplified from Daudi cell mRNA by RT PCR and then cloned into prokaryotic and eukaryotic vectors. The vectors with survivin were transfected into BL21 cells of Escherichia coli and human embryonic kidney 293 cells. The cells were cultured. Protein was extracted from the cells and examined by gel electrophoresis. Suspension of 293 cells was cultured and the number of cells was counted every other day, thus a growth curve was drawn. Another suspension of 293 cells was cultured in soft agar to observe the number of colonies. Cells transfected with plasmids void of survivin were used as controls. Results The anti apoptosis gene survivin was well expressed in BL21 cells and 293 cells. The growth curve showed that the proliferation rate of 293 cells was slightly faster than that of control cells, hoever, without significant difference. Soft agar assay showed that the colonies formed by survivin transfected 293 cells were of greater size and with greater number. Western blotting showed overexpreaaion of cyclin D1 and c myc, two important cancer proteins, in cells transfected with survivin. Conclusion The anti apoptosis gene survivin promotes cell transformation. These effects may depend on the functions of cyclin D1 and c myc.
作者 朱红霞 刘爽 ZHU Hongxia, LIU Shuang, ZHOU Cuiqi, ZHOU Xiaobo, ZHANG Fan, JIN Lanping, XU Ningzhi. Laboratory of Cell and Molecular Biology, Cancer Institute, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China
出处 《中华医学杂志》 CAS CSCD 北大核心 2002年第5期 338-340,共3页 National Medical Journal of China
基金 国家杰出青年基金资助项目( 3992 50 2 0 ) 北京市自然 科学基金资助项目( 7992 0 35)
关键词 基因表达 细胞转化 肿瘤 抗凋亡基因 SURVIVIN Gene expression Cell transformation, neoplastic
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参考文献5

  • 1Ambrosini G,Adida C,Altieri DC.A novel anti-apoptosis gene,survivin, expressed in cancer and lymphoma[].Nature Medicine.1997 被引量:1
  • 2Tamm I,Wang Y,Sausville E,et al.IAP-family protein inhibits caspase activity and apoptosis induced by Fas(CD95), Bax, Caspase,and anticancer drugs[].Cancer Research.1998 被引量:1
  • 3Li FZ,Ambrosini G,Chu EY,et al.Control of apoptosis and mitotic spindle checkpoint by survivin[].Nature.1998 被引量:1
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