Generation of a novel apoptosis-resistant hepatoma cell line.
The expansionable human hepatoma cell lines have potential for use in a bio-artificial liver (BAL) system for liver disease due to the shortage of donation. However, at present, bioartificial livers are incomplete and the functions need to be improved or at least maintained for a longer period. In the present study, the authors aimed to establish a novel hepatoma cell line for a longer-term or permanent artificial liver. For this purpose, bcl-2, an anti-apoptosis gene, was introduced into hepatoma HepG2 cells. Over-expression of Bcl-2 significantly inhibited apoptosis. After 15 d of serum-deprived culture, the viability of HepG2-Bcl2 was 51% while that of mock transfectant (HepG2-mock) was decreased to 14%. In the presence of hygromycin B, HepG2-mock were dead by day 6, while the HepG2-Bcl2 viability at day 9 was 65%. Over-expression of Bcl-2 prolonged the period of the stationary phase in the growth curve and did not affect the growth rate during the exponential phase. To test the liver function, albumin production was measured. After 10 d of culture, the albumin concentration in the culture supernatant of HepG2-Bcl2 was 30 ng ml(-1), while that of HepG2-mock was 23 ng ml(-1). The cytochrome P-450 activity per culture of 3-methyl-cholanthrene-treated HepG2-Bcl2 was double that of treated HepG2-mock. Introduction of Bcl-2 was effective for the generation of a novel hepatoma cell line for artificial livers.
Terada
Satoshi
S
Department of Applied Chemistry and Biotechnology, Faculty of Engineering, Fukui University, 3-9-1 Bunkyo, Fukui 910-8507, Japan. terada@acbio2.fukui-u.ac.jp
Kumagai
Tomoaki
T
Yamamoto
Naohiro
N
Ogawa
Akiko
A
Ishimura
Junichi
J
Fujita
Tetsuo
T
Suzuki
Eiji
E
Miki
Masao
M