Establishment of Transmitochondrial Cell Model for SK-Hep1 mtDNA Depleted Cells and Its Biological Characteristics Analysis

Abstract: To establish ρ°SK-Hep1 cells transmitochondrial model (Cybrids) and analyze mitochondrial DNA (mtDNA) depletion on hepatoma cell's malignant phenotypes. Normal human blood platelets were used as mitochondrial donors， and polyethylene glycol was used as fusion promoting reagent to establish transmitochondrial cell model. These cybrids were confirmed by PCR， Southern hybridization and Western blot. Cells growth status and their invasion capability were detected by MTT assays and Transwell chamber respectively. Western blot were carried out to analyze the expression of apoptosis related protein Bcl-2 and Bax. PCR， Southern hybridization and Western blot confirmed that cybrids cells had objective fragments of mtDNA and positive COX II activity. Cybrids cells showed lower growth rates and less invasive capability than SK-Hep1 and ρ°SK-Hep1 cells. Otherwise， ρ°SK-Hep1 cells had higher growth rates and stronger invasive capability than its parental cells. The ability of apoptotic resistance was highest in ρ°SK-Hep1 cell but reversely in SK-Hep1Cyb cell. ρ°SK-Hep1 cells transmitochondrial model was successfully established. The cybrid cells showed lower growth rates， less invasive capability and decreased anti-apoptotic ability. Damage to mtDNA might have an effect on malignant phenotypes of human hepatoma cells， and those might be reversed by transferring normal mitochondria to the mtDNA depleted cells.
    

CSTR: 32200.14.cjcb.2009.01.0019