Nucleophosmin／B23在肝细胞癌组织中的高表达

High expression of nucleophosmin/B23 in hepatocellular carcinoma

目的探讨Nucleophosmirt／B23(B23)在肝细胞癌(HCC)组织中的表达及其临床病理意义。方法采用重组蛋白表达和杂交瘤细胞技术制备B23重组蛋白和鼠抗B23单克隆抗体。收集103例HCC组织、12例肝局灶性结节性增生和17例肝血管瘤旁肝组织的临床病理档案资料,10例HCC及癌旁肝新鲜组织,采用免疫组织化学(ABC)方法、逆转录聚合酶链反应(RT-PCR)及免疫印迹等技术检测B23在这些组织中的表达,并与增殖细胞核抗原(PCNA)的表达相比较。应用统计学方法对结果进行分析。结果RT—PCR及免疫印迹结果显示:在mRNA和蛋白质水平,B23在HCC组织中的表达明显高于对应的癌旁肝组织。免疫组织化学结果显示:B23在4组(HCC组、癌旁肝组织组、肝局灶性结节性增生组、血管瘤旁肝组织组)中的表达差异有统计学意义(P<0．001),其中B23在HCC组织中的表达显著高于其他3组(P<0．01),PCNA在HCC组织中的表达也显著高于其他3组(P<0．01),相关性分析表明B23与PCNA在4组中表达强度及其差异具有相关性(r=0．4767,P <0．01)。B23在HCC组织中的表达强度与患者血清AFP水平、肿瘤病理分级及是否伴有肝硬化之间的关系有统计学意义(P<0．05)。结论B23在HCC中呈高表达,且显著高于非癌肝组织;B23可作为HCC细胞增生程度的潜在标记,并在临床病理上具有潜在的应用意义。

Objective To study the expression of nucleophosmin/B23 （B23） in tumor cells of hepatocellular carcinoma （HCC） and its clinicopathologic significance. Methods Mouse monoclonal antibodies against B23 were raised by recombinant protein and hybridoma technology. Immunohistochemical study for B23 was performed on 103 cases of HCC, 12 cases of focal nodular hyperplasia and 17 cases of native liver tissue adjacent to hepatic hemangioma. Fresh specimens from 10 cases of HCC and the adjacent liver tissue were also collected for reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blot analysis. The expression of B23 was analyzed and compared with that of proliferative cell nuclear antigen （PCNA） in these specimens. Results RT-PCR and Western blot analysis showed that B23 expression in HCC was higher than that in adjacent liver tissue. Statistically significant difference in expressions of B23 and PCNA were also noted in the four groups studied （P 〈 0.01）. B23 and PCNA expressions in HCC were higher than those in the other three groups （P 〈 0.01）. There was also a statistically significant correlation between B23 and PCNA expressions amongst the four groups （r = 0.4769, P 〈 0.01）. Besides, B23 expression in HCC correlated with pathologic tumor grading, serum alpha-fetal protein levels and cirrhotic status （P 〈 0.05）. Conclusions B23 expression in HCC was significantly higher than that in liver tissues with non-malignant diseases. B23 may be used as a marker for neoplastic changes in liver cells and thus has potential clinicopathologic application.