中国肿瘤生物治疗杂志2011，Vol.18Issue(4)：383-388,6.DOI:10.3872/j.issn.1007-385X.2011.04.008
乳腺癌特异性多肽介导的HSV-TK/GCV系统的构建及其靶向杀伤效应
Construction and targeted effects of breast cancer specific peptide-mediated HSV-TK/GCV anti-tumor system
摘要
Abstract
Objective: To investigate the targeted killing effect of breast cancer specific peptide PI-mediated HSV-TK/ GCV system against human breast cancer MDA-MB-231 cells in vitro. Methods: PI-TK gene was amplified from pORFHSV-TK plasmid by PCR, and was re-inserted into the prokaryotic expression plasmid pET-28a ( + ) ; the pET-28a ( + ) -PI-TK plasmid was constructed and transfected into the host bacteria. After induction with IPTG, PI-TK fusion protein was purified by His-Tag and further identified by SDS-PAGE and Western blotting analysis. MDA-MB-231 cells were cultured with different dosages of PI-TK fusion protein; after further treatment with ganciclovier (GCV) , the morphology changes of MDA-MB-231 cells were observed under inverted microscope, and the cell proliferation was evaluated by CCK8 assay. Results:Recombinant prokaryotic expression plasmid pET-28a ( + )-PI-TK was successfully constructed. The purified PI-TK fusion protein was obtained and confirmed by the SDS-PAGE and Western blotting analysis. PI-TK fusion protein alone failed to affect the morphology and proliferation of MDA-MB-231 cells, but when combined with GCV, PITK fusion protein specifically inhibited the proliferation of MDA-MB-231 cells in a dose-dependent manner, with inhibitory rate of 200 |xg/ml PI-TK + 10 mg/L GCV being (68.9 ±7.57)% , and IC50 being 152.64 μg/ml, but it had no effects on MDA-MB435 cells (P＜0.05). Conclusion; The breast cancer specific peptides Pi-mediated HSV-TK antitumor system can specifically kill MDA-MB-231 cells.关键词
乳腺癌特异性多肽/PI/乳腺癌/MDA-MB-231细胞/HSV-TK/GCV抗瘤系统/靶向性Key words
breast cancer specific peptide/ breast cancer/ MDA-MB-231 cell/ HSV-TK/GCV suicide gene system/ targeted分类
医药卫生引用本文复制引用
耿计伟,董坚,刘为青,高嫦娥,熊秋霞,缪延栋,周华华,李秋恬,李臣..乳腺癌特异性多肽介导的HSV-TK/GCV系统的构建及其靶向杀伤效应[J].中国肿瘤生物治疗杂志,2011,18(4):383-388,6.基金项目
国家自然科学基金资助项目(No.3086033) （No.3086033）
云南省应用基础研究重点项目资助(No.2009CC023):云南省科技基础条件平台建设项目资助(No.2007DA006) （No.2009CC023）
