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KMID : 0360319950270040559
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Journal of Korean Cancer Research Association
1995 Volume.27 No. 4 p.559 ~ p.569
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Effects of Retinoic Acid on p53 Protein, Ki-67 and PCNA/Cyclin Eepression in PLC/PRF/5 Cells
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김대곤
권철/류완희/김이엽/안득수
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Abstract
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Retinoic acid(RA), known to have antiproliferative and differentiation-inducing effects on cancer cells, was examined to evaluated its potential as a chemotherapeutic agent for hepatocellular carcinoma.
The treatment of PLC/PRF/5 cells with RA(10μM for 8 days) resulted in inhibited growth by 23.8% as compared to that of control. The decreased cell growth rate was started 2 days after RA addition.
We examined the level of expressions of the mutated p53 protein, the Ki-67 antigen, and the proliferation cell nuclear antigen(PCNA)in cultured PLC/PRF/5 hepatocelluar carcinoma cells before and after RA treatment. The mutated p53 is though to be
involved ir oncogenesis of human cancers, and the expression of the Ki-67 and the PCNA are used to evaluate tumor cell kinetics.
The expression of p53 protein was ot inhibited significantly in PLC/PRF/5 cells by treatment with 10-5 M RA in G0/G1, S, or G2/M phase fraction, as detected by immunocytochemical staining using the monoclonal antibody Pab 180 which recognizes
both
the
wild and the mutated p53 protein. Also no significant increase of expression was observed using the monoclonal antibody Pab 240 which binds only the mutated p53 protein.
RA treatment increased G0 nustained fraction from 0.36±0.06% to 0.7±0.19% and decreased Ki-67 antigen expression significantly from 51.2±0.8% to 46.9±0.4%(p=0.03) in G1phase, 14.8±0.5% to 11.3±0.5%(p=0.03) in S phase respectively, but
increased
the fraction of G2/M phase from 33.4±1.1% to 36.8±1.4%(p=0.02).
PCNA expression was also dropped by RA treatment from 50.4±1.5% to 42.8±0.3%(p=0.01) in G0/G1 fraction, 14.7±0.5% to 11.9±0.4%(p=0.02) in S phase, increased G2/M phase fraction from 31.3±1.2% to 42.2±0.6%(p=0.008).
These results suggested that RA may have anticancer effect through the inhibition of Ki-67 and PCNA expression in similar cell cycle phase without affecting the expression of mutant p53 protein in PLC/PRF/5 cells was observed. In addition, cell
cycle
analysis suggested that RA induced an arrest of G0 progression to G1 and S phase.
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