Inhibition of human endothelial cell proliferation by Gami--Jeonggi--San (Jiawei--Zhenqi--San) is accompanied by transcriptional up-regulation of p53 and Waf1 tumor suppressor genes.
The effects of Gami-Jeonggi-San (GJS) on proliferation of human endothelial cell (HUV-EC-C) were investigated using a flow cytometry and a quantitative RT-PCR analysis of gene expression. An accumulation of cells at G(1) phase of the cell cycle was found at 72 h after treatment (10 microl/ml) while no detectable reduction of PCNA expression was recognized. To elucidate that the cell cycle inhibitory effect of GJS stems from its capability of transcriptional regulation of the cell cycle-controlling genes, we investigated mRNA expression of p53, Waf1, PCNA, Cyclin D1, Cdc2, Histone H3, c-Myc, and c-Fos. Significantly elevated mRNA levels of the p53 tumor suppressor gene and its down-stream mediator gene, Waf1, whose increased expressions were known to trigger G(1) cell cycle arrest, were observed. In contrast, a marked reduction of two early G(1)-specific, cell cycle stimulating genes, c-Myc and c-Fos, were found at 24h after treatment, while there were no detectable changes in expressions of G(1)-S or G(2)-M transition-related genes, indicating the G(1) specificity of GJS effect on the cell cycle. These results suggest that the pharmacological effects of GJS might be derived in part from inhibition of cellular proliferation of human endothelial cells, and that GJS inhibition of the cell cycle might stem from its regulatory capability on the transcription of the cell cycle-controlling genes, including p53 and Waf1 tumor suppressor genes.
Moon
Sang-Kwan
SK
Department of Circulatory Internal Medicine, College of Oriental Medicine, Kyung Hee University, Seoul 130-702, Republic of Korea.
Jin
Hyung-Seok
HS
Ko
Chang-Nam
CN
Kim
Young-Suk
YS
Bae
Hyung-Sup
HS
Lee
Kyung-Sup
KS
Cho
Ki-Ho
KH
D002453:Cell Cycle / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000235:genetics; D002460:Cell Line; D050759:Cyclin-Dependent Kinase Inhibitor p21; D042783:Endothelial Cells / Q000187:drug effects*; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects*; D016147:Genes, Tumor Suppressor / Q000187:drug effects*; D016158:Genes, p53 / Q000187:drug effects; D006801:Humans; D008516:Medicine, Chinese Traditional; D008518:Medicine, East Asian Traditional; D012333:RNA, Messenger / Q000378:metabolism; D015854:Up-Regulation
C493956:CDKN1A protein, human; D018797:Cell Cycle Proteins; D050759:Cyclin-Dependent Kinase Inhibitor p21; D012333:RNA, Messenger