Department of Biochemistry and Molecular Biology, College ofMedicine, Yeungnam University, Daegu, Korea. jhykim@med.yu.ac.kr
Published online: October 31, 2001.
ABSTRACT
PURPOSE: It has been demonstrated that PLC-gamma1 is overexpressed in many tumor cells, and that overexpression of Phospholipase C (PLC)-gamma1 is associated with tumor progression. In order to understand the effect of the PLC-gamma1 overexpression on the regulation of cell cycle regulators following DNA damage, we analyzed the expression level of PCNA, cyclin B1, and p21 Waf1 after ultraviolet C (UVC) irradiation in PLC-gamma1-transfected PC12 cells.
MATERIALS AND METHODS: PC12 and 3Y1 cells, transfected with empty vector or rat PLC-gamma1 cDNA, were used for this study. Following UVC irradiation, cell cycle progression was analyzed by flow cytometry and protein expression was detected by Western blotting.
RESULTS: Waf1 protein was markedly down-regulated, whereas PCNA and cyclin B1 was up-regulated in PLC-gamma1 overexpressed-cells as compared to the vector transfected-cells. When the cells were irradiated with UVC, PCNA was slightly increased within 3-hours of the UV irradiation and then was markedly decreased in Vector/ PC12 cells, while it remained high until 37 hour after UVC in PLC-gamma1/PC12 cells. In contrast, cyclin B1 was gradually decreased following UVC irradiation in both cells.
CONCLUSION: The overexpression of PLC-gamma1 affects the expression level of PCNA after UVC irradiation. We proposed that the overexpression of PLC-gamma1 may contribute to the UV-induced genomic instability by up-regulating the expression of PCNA.