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J Korean Cancer Assoc > Volume 27(4); 1995 > Article
Journal of the Korean Cancer Association 1995;27(4): 559-570.
PLC / PRF / 5 간암세포주에서 Retinoic Acid 가 p53 변이 단백 , Ki - 67 및 PCNA / Cyclin 의 발현에 미치는 효과
김대곤, 권철, 류완희, 김이엽, 안득수
Effects of Retinoic Acid on p53 Protein , Ki - 67 and PCNA / Cyclin Expression in PLC / PRF / 5 Cells
Dae Gon Kim, Cheol Kwon, Wan Hee Yoo, Yee Yup Kim, Deuk Su Ahn
ABSTRACT
Retinoic acid(RA), known to have antiproliferative and differentiation-inducing effecte on cancer cells, was examined to evaluate its potential as a chemotherapeutic agent for hepatocellular carcinoma. The treatment of PLC/PRF/5 cells with RA(l0 pM 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 pM is thought to be involved in oncogenesis of human cancers, and the expressions of the Ki-67 and the PCNA are used to evlauate tumor cell kinetics. The e#xpression of p53 protein was not inhibited significantly in PLC/PRF/5 cells by treatment with 10 M RA in Go/Gi, S, or G/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 increaaed Go unstained fraction from 0.36+-0.06% to 0.7+-0.19% and decreased Ki-67 antigen expresaion significantly from 51.2+0.8% to 46.9+0.4%(p=0.03) in G phase, 14.8 0.5% to 1l.30.5%(p=0.03) in S phase respectively, but increased the fraction of GgM 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 Gw G, fraction, 14.7+-0.5% to 11.9+-0.4%(p=0.02) in S phase, increased G/M phase fraction from 31.3+-l.2% to 42.2+-0.6%(p=0.008). These results suggested that RA may have anticancer effect through the inhibitiion 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 anaysis suggested that RA induced en arrest of G, progression to G, and S phase.
Key words: Retinoic acid(RA), PLC/PRF/5 cells, p53, Ki-67, PCNA
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