Construction of mitochondria targeted parvalbumin expressing vector and its effect on mitochondrial calcium homeostasis of hepatocellular carcinoma cells
ZHANG Hui
WANG Jiaojiao
REN Tingting
JIN Mingpeng
ZHU Jianjun
ZHANG Hongxin
WU Yousheng
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 .
Keywords：liver neoplasmsparvalbuminmitochondriacalcium homeostasisvector construction
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 4-6 )