重组腺病毒Ad—GFP—C197对3种肿瘤细胞的抑制作用研究
邬贤 黄莹 李舒婕 段萍萍 米鹏程 陈文瑛中图分类号 R966;R979.1 文献标志码 A 文章编号 1001-0408(2018)20-2800-05
DOI 10.6039/j.issn.1001-0408.2018.20.14
摘 要 目的:研究以腺病毒载体系统装载人端粒酶逆转录酶C端片段(C197)制成的重组腺病毒Ad-GFP-C197对3种肿瘤细胞增殖的抑制作用。方法:采用HEK293细胞扩增并纯化Ad-GFP-C197。采用Ad-GFP-C197分别感染人胃癌细胞SGC7901、人乳腺癌细胞MCF7和人结直肠癌细胞CaCO2。以空白腺病毒载体(Ad-GFP)为参比,采用Western blot法检测Ad-GFP-C197感染后3种肿瘤细胞中C197的蛋白表达水平;采用MTT法检测Ad-GFP-C197感染后对3种肿瘤细胞的抑制作用,绘制细胞增殖曲线并计算增殖抑制率。结果:Ad-GFP感染的3种肿瘤细胞中均未检测到C197蛋白表达,而Ad-GFP-C197感染的上述细胞中C197蛋白均有明显表达;Ad-GFP-C197感染后3种肿瘤细胞的增殖曲线随时间延长而呈明显抑制现象,其增殖抑制率最高可达37.31%~41.42%。结论:Ad-GFP-C197对SGC7901细胞、MCF7细胞、CaCO2细胞增殖均有明显的抑制作用,且起效迅速,能够克服其他端粒酶抑制剂起效缓慢的缺点。
关键词 人端粒酶逆转录酶;端粒酶抑制剂;重组腺病毒;Ad-GFP-C197;肿瘤细胞;细胞增殖
ABSTRACT OBJECTIVE: To study the inhibitory effects of recombinant adenovirus Ad-GFP-C197, which prepared by adenovirus vector system-loading human telomerase reverse transcriptase (hTERT) C fragment (C197), on the proliferation of 3 kinds of tumor cells in vitro. METHODS: Ad-GFP-C197 was amplified and purified with HEK293 cells. Human gastric cancer cells SGC7901, human breast cancer cells MCF7 and human colorectal cancer cells CaCO2 were infected by Ad-GFP-C197 respectively. Using blank adenovirus carrier (Ad-GFP) as reference ......
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