原发性肝细胞癌中WAF_1/CIP_1基因的表达及其与p53基因突变的关系

WAF1/CIP1 gene expression in human hepatocellular carcinoma and its relationship with p53 mutation

目的研究原发性肝细胞癌（ＨＣＣ）组织中ＷＡＦ１基因的表达及其与ｐ５３基因突变和临床病理学指标的关系和意义。方法应用半定量ＲＴ－ＰＣＲ、免疫组织化学及定量ＤＮＡ图像分析的方法，检测ＷＡＦ１基因在３２例ＨＣＣ及其癌旁肝组织和５例正常肝组织中的表达，研究其与ｐ５３基因突变及临床病理学指标的关系。结果肝癌组织中ＷＡＦ１ｍＲＮＡ表达相对值（１０６１０．３７）Ｕ低于其相应癌旁肝组织（１３０±０．３７）Ｕ，Ｐ＜０．０１，Ｐ２１ｗＡＦ１表达阳性率（５３．１％）明显低于癌旁肝组织的门００％，Ｐ＜０．０１）。有卫星灶形成的肝癌ＷＡＦｌｍＲＮＡ的表达（０．９２±０．３３）Ｕ明显低于无卫星灶形成的肝癌（１．２４±０．３５）Ｕ，Ｐ＜０．０１，且ｐ２１ｗＡＦ嚷达阳性率（３３．３％）低于无卫星灶形成的肝癌（７８．６％，Ｐ＜０．０５）。存在Ｐ５３基因突变的肝癌，其ＷＡＦ１ｍＲＮＡ的表达水平（０．９２±０．２８）Ｕ低于无ｐ５３基因突变的肝癌（１．１６±０４０）Ｕ，Ｐ＜０．０５，且ｐ２１ｗＡＦ１表达阳性率（１５．４％）显著低于无ｐ５３基因突变的肝癌（７．９％，Ｐ＜０．０１）。在Ｐ２１ｗＡＦ１表达阳性的肝癌中处于Ｓ期的细胞数（４４．５７±７．５?

Objevtive To study the WAF1 gene expression in human hepatocellular carcinoma(HCC)and its relationship with p53 mutation. Methods Using semiQuantitative reverse tra use riptase - polymerasechain reaction(RT-PCR) and immunohistochemistry, the expression of WAFl and p53 in 32 HCC samplesand their surrounding liver tissues and 5 normal liver tissues was detected respectively. Results Theexpression level of WAFl mRNA in HCC samples was lower than that in paratumor liver tissues(l.06 Ut 0.37 U vs l.30 U 0.37 U, P< 0.01) and its positive rate of p21'wAF was also lower than that inparatumor liver tissues(53.l% vs 100%, p<0.01). The expression level of WAFl mRNA in HCC samples withintra-hepatic metastatic lesions was lower than that without intra-hepatic metastatic lesions(0.92 U 0.33 Uvs l.24 U 0.35 U, p<0.01) and its positive rate of p21wAF1 was also lower than that without intra-hepaticmetastatic lesions(33.3% vs 78,6%, P<0.05). The expression level of WAF1 mRNA in HCC samples withp53 mutation was lower than that without p53 mutation (0.92 U 0.28 U vs l.16 U 0.40 U, P< 0.05) andits positive rate of p21wAF was also lower than that without p53 mutation(15.4% vs 78.9%, p<0.01). Thepercentagr of cells in S-phase in p21'wAF1-positive HCC samples was lower than that in p21wAFl-negstiveHCC samples(44.57% 7.56% vs 51.83% 8.51%, P< 0.01). However, no significant difference in expressionof WAFl mRNA and p21wAF1 in HCC samples was found in other clinical pathological parameters such asthe degree of differentiation, the histological pattern, the size of tumors, the status of capsule in tumorsand the formation of tumor thrombus of portal vein. The expression level of WAFl mRNA in normalimmunohistochemical staining of p21'AP1 in allnormal liver tissues was positive(100%). Conclusion The expression of WAFl gene in HCC is predominantly regulated by dependence on p53 and the reduced WAFl expression may Darticipate in thecareing6nesis and progression of HCC and the mechanisms of infiltration and metastasis of HCC.