hTERT启动子调控rVBMDMP基因表达载体的构建与功能研究
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湖南省自然科学基金(07JJ6164)和湖南省重点学科建设基金(2006-180)资助项目


Construction and Function Study of Expression Vector of rVBMDMP Gene Regulated by hTERT Promotor
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This work was supported by grants from The Hunan Provincial Natural Science Foundation of China (07JJ6164) and The Construct Program of The Key Discipline in Hunan Province (2006-180)

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    摘要:

    为了探讨端粒酶催化亚单位(hTERT)启动子调控重组血管基膜衍生多功能肽(rVBMDMP)基因表达抑制肺癌细胞生长的作用机制,采用PCR方法,克隆hTERT启动子核心区片段,并检测其功能活性.然后,构建pLNSX/hTERT/rVBMDMP逆病毒载体,获取逆病毒,感染肺癌A549细胞,检测hTERT启动子调控rVBMDMP基因表达对细胞形态、细胞生长、细胞凋亡以及Caspase-3表达的影响.另外,观察hTERT启动子调控rVBMDMP基因表达对裸鼠成瘤的抑制作用、瘤组织细胞的凋亡及Caspase-3表达情况.结果发现:a.hTERT启动子核心区能调控rVBMDMP基因的表达(n=3,P < 0.05);b.hTERT启动子调控rVBMDMP基因的表达具有抑制肺癌A549细胞的生长和裸鼠成瘤作用(n=6,P < 0.05);c.rVBMDMP基因表达能促进肺癌A549细胞凋亡和Caspase-3蛋白表达.以上结果说明,hTERT启动子调控rVBMDMP基因表达后,通过提高Caspase-3的表达水平,促进细胞凋亡而抑制肺癌A549细胞的生长.

    Abstract:

    To investigate action mechanism of the expression of rVBMDMP gene regulated by hTERT promotor inhibited cell growth of lung cancer, the Core domain of hTERT promotor was obtained by PCR and its function activity was detected. Then, retroviral vector of pLNSX/hTERT/rVBMDMP was recombined, and lung cancer A549 cells were infected by the collected vetrovirus. Further, it was observed that expression of rVBMDMP gene regulated by hTERT promotor influenced cellular morphous, cell growth, cell apoptosis and expression of Caspase-3 protein. Moreover, it was observated that expression of rVBMDMP gene regulated by hTERT promotor inhibited the growth of transplanted tumor in the nude mice, the cellular apoptosis of tumor tissue and the expression of Caspase-3 protein. The results showed: a. The Core domain of hTERT promotor can regulate the expression of rVBMDMP gene (n=3, P < 0.05); b. Expression of rVBMDMP gene regulated by hTERT promotor was abled to inhibit the growth of lung cancer A549 cells and transplanted tumor in nude mice (n=6, P < 0.05); c. The expression of rVBMDMP gene promoted the apoptosis of lung cancer A549 cells and the expression of Caspase-3 protein. These results suggested that expression of rVBMDMP gene regulated by hTERT promotor inhibited the growth of lung cancer A549 cells because it enhanced expression level of Caspase-3 protein to increase cell apoptosis.

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张志伟,贺修胜,罗招阳,李艳兰,曹建国. hTERT启动子调控rVBMDMP基因表达载体的构建与功能研究[J].生物化学与生物物理进展,2010,37(4):416-425

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  • 收稿日期:2009-08-24
  • 最后修改日期:2010-01-24
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  • 在线发布日期: 2010-02-04
  • 出版日期: 2010-04-20