Establishment of Human Hepatocellular Cancer Cell Line Lacking Mitochondrial DNA

Abstract: To establish a human hepatocellular cancer cell line lacking mitchondrial DNA (mtDNA). Human hepatocellular cancer cell line SK-Hep1 was treated by ethidium bromide (EB)， and to verify the cell line lacking mtDNA， the cells were cultured in media without uridine and pyruvate， and PCR and southern hybridization were used to detect cytochrome oxidase subunit I (COXI) and II (COXII). After exposure to EB for 1 day， some SK-Hep1 cells began swelling and suspending， almost all cells died for about 10?2 days when cultured in the media without uridine and pyruvate. When SK-Hep1 cells were cultured in the media with EB and uridine and pyruvate for 21 generations， COXI and COXII cannot be amplified， and Southern blot suggested there were not COXI and COXII hybridization bands. Treated By EB， we have successfully established a human hepatocellular cancer cell line lacking mtDNA (ρ⁰SK-Hep1)

CSTR: 32200.14.cjcb.2007.01.0025