山东医药2016，Vol.56Issue(9)：4-6,3.DOI:10.3969/j.issn.1002-266X.2016.09.002
线粒体靶向小清蛋白表达载体构建及其对肝癌细胞线粒体钙稳态的影响
Construction of mitochondria targeted parvalbumin expressing vector and its effect on mitochondrial calcium homeostasis of hepatocellular carcinoma cells
摘要
Abstract
Objective To construct mitochondria targeted parvalbumin ( PVALB) expression vector and to investigate the mitochondrial targeting PVALB expression in human hepatocellular carcinoma cells MHCC 97H and its effect on mito-chondrial calcium homeostasis .Methods Using skeletal muscle cells cDNA as the template , the PVALB was amplified by PCR, and then the digested PVALB was connected with MTS and pcDNA 3.1 (+) vector.After identifying by enzyme di-gestion analysis and sequencing , the recombinant plasmid was transfected into MHCC 97H by liposome.The expression and localization of PVALB, calcium concentration in MHCC97H were detected by laser scanning confocal microscopy after transfection.Results pcDNA3.1 (+)-MTS-PVALB was successfully constructed and was verified by restriction enzyme digestion and DNA sequencing .The expression of MTS-PVALB was located in the mitochondria , and MTS-PVALB could down-regulate the concentration of calcium .Conclusion pcDNA3.1 ( +)-MTS-PVALB was successfully constructed . The expressed MTS-PVALB was located in MHCC97H mitochondria and regulated mitochondrial calcium homeostasis .关键词
肝肿瘤/小清蛋白/线粒体/钙稳态/载体构建Key words
liver neoplasms/parvalbumin/mitochondria/calcium homeostasis/vector construction分类
医药卫生引用本文复制引用
张卉,王姣姣,任婷婷,金明朋,朱剑军,张洪新,吴有盛..线粒体靶向小清蛋白表达载体构建及其对肝癌细胞线粒体钙稳态的影响[J].山东医药,2016,56(9):4-6,3.基金项目
国家自然科学基金资助项目(81572727 （）
81572304). （）
