Folate Deficiency Induces a Cell Cycle-Specific Apoptosis in HepG2 Cells

Manuscript received 8 July 1998. Initial reviews completed 23 July 1998. Revision accepted 11 September 1998.

Rwei-Fen S. Huang^*, Yun-Hsiu Ho^*, Huei-Li Lin^*, Jeng-Shu Wei, and Tsan-Zon Liu^**,

^* Department of Nutrition and Food Sciences, Fu-Jen University, Hsin-Chuang, Taiwan,  Graduate Institute of Basic Medicine, School of Medical Technology, Chang Gung University, Taoyuan, Taiwan and ^** Department of Medical Research, Yuan's General Hospital, Kaohsiung, Taiwan, R.O.C.

The human hepatoma HepG2 cell line was chosen as a representative of solid tissue-derived cell systems in which folate metabolism and apoptosis induction have not been thoroughly investigated. HepG2 cells were cultivated in the control or folate-deficient media (control media lacking of folate, glycine, thymidine and hypoxanthine) for 4 wk. This resulted in a decrease in intracellular folate levels to 32% of the control within 1 wk, which was followed by growth arrest and greater cell death rates. These disturbances of folate deficiency coincided with apoptotic induction, as characteristically shown by nucleosomal DNA fragmentation of 180-200 base pair multimers, nuclear chromatin condensation and positive terminal transferase-mediated dUTP nick end labeling assay. Apoptosis coincided with an accumulation of cells in S-phase, a subsequent G2/M phase block and a significant increase in mean protein content as evaluated by flow cytometric analyses employing a double-staining method. The growth and cell cycle arrest under folate-deficient conditions was independent of a change of p53 expression as measured by an enzyme-linked immunosorbent assay. Supplementation of 2 µmol/L folate normalized cell cycles and diminished DNA fragmentation. Taken together, these data indicate that HepG2 cells cultivated in folate-deficient medium have a low folate concentration, decreased growth and viability, and increased apoptotic propensity. This occurrence of apoptosis was associated with a cell cycle-specific mechanism and independent of p53-mediated pathway.

The Journal of Nutrition Vol. 129 No. 1 January 1999, pp. 25-31
Copyright ©1999 by the American Society for Nutritional Sciences

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