[The killing effect and bystander effect of linamarase/linamarin suicide gene system on human hepatocellular carcinoma cell line HepG2 in vitro].

Abstract

AIM:

To investigate the killing effect of linamarase/linamarin (lis/lin) system on hepatocellular carcinoma cell line HepG2 in vitro.

METHODS:

A cDNA library was built from RNA of cassava by RT-PCR, then the linamarase gene was amplified from it by PCR and cloned into the eukaryotic expression vector plasmid pEGFP-N1, which made up the recombinant plasmid pEGFP-N1-lis. The human HCC cells HepG2 were transfected with the recombinant plasmid mediated by electroporation and screened by G418 to yield the positive clone which was termed HepG2/lis. The expression of lis was confirmed by fluorescent staining, RT-PCR and Western blot. The killing effect and bystander effect of linamarin with different concentrations on HepG2 was detected by MTT.

RESULTS:

RT-PCR confirmed the expression of lis gene in HepG2 and Western blot analysis confirmed existence of lis-EGFP fusion protein in HepG2. Linamarin in low concentration had shown notable cytotoxic effect on HepG2/lis. When HepG2/lis cells were mixed with parental HepG2 cells at a ratio of 10:90 and cultivated in 500 mg/L lin medium, significant bystander effect was observed in vitro.

CONCLUSION:

The linamarase/linamarin suicide gene system has strong killing effect and bystander effect on HCCs with the concentration of 500 mg/L lin.

• Authors: Ming-hao Chen, Jun Ma, Hai-min Li, Jun Li, Ren Li, Fu-qin Zhang, Ke-feng Dou
• Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology (Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi) Vol. 24 Issue 3 Pg. 274-7 (Mar 2008) ISSN: 1007-8738 [Print] China
• PMID: 18328192 (Publication Type: English Abstract, Journal Article)

Chemical References

• Nitriles
• cyanogenic beta-glucosidase
• beta-Glucosidase
• linamarin

Topics

• Blotting, Western
• Bystander Effect (physiology)
• Carcinoma, Hepatocellular (therapy)
• Genes, Transgenic, Suicide (genetics, physiology)
• Genetic Therapy (methods)
• Hep G2 Cells
• Humans
• Liver Neoplasms (therapy)
• Nitriles (metabolism)
• Reverse Transcriptase Polymerase Chain Reaction
• beta-Glucosidase (genetics, physiology)