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机构地区:[1]北京医科大学生化与分子生物学系,100083
出 处:《中华病理学杂志》2000年第3期188-191,共4页Chinese Journal of Pathology
基 金:国家自然科学基金资助课题!(39930170);国家重点基础研究发展规划!(G1999053906)资助课题
摘 要:目的 研究端粒酶RNA的反义对人乳腺癌细胞系MCF 7细胞端料酶活性的影响。方法 用重组腺病毒转移并表达端粒酶RNA的反义cDNA ,采用基因重组、脂质体共转染法获得端粒酶反义重组病毒 ,用Southern杂交鉴定病毒的整合功能 ,用TRAP 银染法检测端粒酶活性。结果 MCF 7细胞是恶性乳腺癌的典型细胞系。与对照组MCF 7、MCF 7 vAd AAV细胞相比 ,反义病毒感染后的MCF 7 vAdT AAV细胞的端粒酶活性明显降低 ,只扩增出小片段的寡聚核苷酸。结论 端粒酶RNA的反义cDNA的导入 。Objective To inhibit telomerase activity in breast cancer cells MCF 7 by antisense ribonucleotide technique. Methods Recombinant adenovirus was chosen as gene delivery vector to transfer and express the antisense RNA component of telomerase in tumor cells. The recombinant adenovirus of anti telomerase RNA was obtained through gene recombination and co transfection with lipofectin. The integrated potential of antisense virus was certified by Southern blot and the telomerase activity of MCF 7 was analysed by TRAP silver staining. Results The MCF 7 cell line is a typical model for malignant breast cancer. In this study, the telomerase activity of MCF 7 was significantly inhibited after antisense virus infection when compared with the control cell lines. Conclusion The introduction of antisense cDNA of telomerase RNA can significantly inhibit the activity of telomerase in MCF 7 cells.
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