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逆转录病毒介导重组CXCR4对前列腺癌细胞侵袭力的影响 被引量:1

Effects of retroviral-mediated recombinant CXCR4 gene on invasion of prostate cancer cell line in vitro
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摘要 目的:构建CXCR4基因重组逆转录病毒表达载体pLEGFP-CXCR4,并转染PC-3细胞,探讨趋化因子受体CXCR4对人前列腺癌细胞体外侵袭能力的影响。方法:分离健康人外周血单个核细胞,提取总RNA,以其为模板,RT-PCR法扩增CXCR4,获取CXCR4基因全长序列,装入带有绿色荧光蛋白(GFP)的逆转录病毒质粒pLEGFP-N1,用脂质体法转染PC-3细胞,采用实时定量PCR和Western blotting观察CXCR4表达情况,并通过体外细胞-基质粘附试验和Transwells小室检测肿瘤细胞体外侵袭能力的变化。结果:成功构建pLEGFP-CXCR4载体,转染人前列腺癌细胞72h后,CXCR4 mRNA及蛋白质水平明显上调,细胞的体外侵袭力明显增强,增殖活性增强。结论:CXCR4转染能够明显提高CXCR4基因mRNA及蛋白水平,提高人前列腺癌细胞体外侵袭能力。 Objective:To constructing CXCR4 gene recombinant retrovirus vector PLEGFP - N1, then transfect PC -3 cells, and study the effect of chemokine receptor CXCR4 - targeted on invasion capability of human prostate Cancer cell line in vitro. Methods:Total RNA was extracted from monocyte separated from peripheral blood of health human. And CXCR4 gene was amplified by RT- PCR,then inserted it into retroviral vector which contain Green Flu Protein, named pLEGFP- N1. Then transfected with lipofectamine mediation to PC -3 cells. The mRNA level of CX- CR4 was detected by real - time PCR, expression of CXCR4 protein was detected by Western blotting at 72h after transfection. The abilities of invasion were detected by Matrigel invasion assay. Results :The retrovirus vectors pLEG- FP - CXCR4 were constructed successfully, the expression of CXCR4 mRNA and protein was up - regulated significantly after transfection of CXCR4. The invasion and proliferation capabilities of PC - 3 cells were enhanced. Conclusion: CXCR4 effectively up - regulates the expression of CXCR4 gene and improves invasion capability of PC - 3 cells in vitro.
出处 《现代肿瘤医学》 CAS 2010年第2期213-216,共4页 Journal of Modern Oncology
基金 国家自然科学基金面上项目(编号:30500504) 第三军医大学校管课题(编号:XG200533)
关键词 前列腺癌 CXCR4 逆转录病毒载体 肿瘤侵袭 prostate cancer CXCR4 retrovirus vector neoplasm invasiveness
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  • 1Jun Hayakawa,Makoto Migita,Takahiro Ueda,Takashi Shimada,Yoshitaka Fukunaga.Generation of a Chimeric Mouse Reconstituted with Green Fluorescent Protein-Positive Bone Marrow Cells: A Useful Model for Studying the Behavior of Bone Marrow Cells in Regeneration In Vivo[J].International Journal of Hematology.2003(5)

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