• 동의생리병리학회지
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• 제22권4호
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• pp.821-828
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• 2008

Onchungeum, a herbal formula, which has been used for treatment of anemia due to bleeding, discharging blood and skin disease. In the present study, it was examined the effects of extract of Onchungeum (OCE) on the growth of human hepatocarcinoma cell lines Hep3B (p53 null type) and HepG2 (p53 wild type) in order to investigate the anti-proliferative mechanism by OCE. Treatment of Hep3B and HepG2 cells to OCE resulted in the growth inhibition in a dose-dependent manner, however Hep3B cell line exhibited a relatively strong anti-proliferative activity to OEC. Flow cytometric analysis revealed that OCE treatment in Hep3B cells caused G1 phase arrest of the cell cycle, which was associated with various morphological changes in a dose-dependent fashion. RT-PCR and immunoblotting data revealed that treatment of OCE caused the down-regulation of cyclin D1 expression, however the levels of cyclin E expression were not changed by OCE. The G1 arrest of the cell cycle was also associated with the induction of Cdk inhibitor p27 by OCE. Because the p53 gene is null in Hep3B cells, it is most likely that the induction of p21 is mediated through a p53-independent pathway. Moreover, p27 detected in anti-Cdk4 and anti-Cdk2 immunoprecipitates from the OCE-treated cells, suggesting that OCE-induced p27 protein blocks Cdk kinase activities by directing binding to the cyclin/Cdk complexes. Furthermore, OCE treatment potently suppresses the phosphorylation of retinoblastoma proteins and the levels of the transcription factor E2F-1 expression. Taken together, these results indicated that the growth inhibitory effect of OCE in Hep3B hepatoma cells was associated with the induction of G1 arrest of the cell cycle through regulation of several major growth regulatory gene products.

• 동의생리병리학회지
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• 제22권5호
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• pp.1250-1255
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• 2008

Onchungeum(OCE), a herbal formula, has been used for treatment of anemia, discharging blood and skin diseases. In the previous study, we investigated the anti-cancer effect of OCE by G1 arrest of the cell cycle in human hepatocarcinoma cells, Hep3B cells. In this study, it was examined that the difference of anti-proliferative mechanisms by change in the ratio of OCE composition (OCE I) in Hep3B cells. Treatment of OCE I exhibited a relatively strong anti-proliferative activity and caused various morphological changes such as membrane shrinkage and cell floating. In addition, OCE-I arrests the cell cycle at G1 phase, which was associated with the down-regulation of cyclin D1 and Cdk6 expressions. The G1 arrest was also associated with the induction of Cdk inhibitors p27 and p21. Moreover, both p21 and p27 were detected by immunoprecipitation with anti-Cdk4 and anti-Cdk2 antibodies in OCE I-treated cells but in case of OCE, p21 did not make any complexes with Cdk4 and Cdk2. These results suggest that the change in the ratio of OCE composition might induce different mechanisms in anti-cancer efficacy of OCE, which may confer characteristic principles in oriental medical formula.