[应用组织芯片技术检测8种p53相关基因在肝癌中的表达]

[Determination of expression of eight p53-related genes in hepatocellular carcinoma with tissue microarrays].

作者信息

Zhu Ming-Hua, Ni Can-Rong, Zhu Zhi, Li Fang-Mei, Zhang Shun-Min

机构信息

Department of Pathology, Changhai Hospital, Second Military Medical University, Shanghai, 200433, PR China.

出版信息

Ai Zheng. 2003 Jul;22(7):680-5.

PMID:12866955

Abstract

BACKGROUND & OBJECTIVE: Hepatocarcinogenesis was a multistage process involving a number of genes. Molecular biological studies indicated that abnormal expression of p53 gene family is correlated with the development of hepatocellular carcinoma (HCC). This study was designed to investigate the expression difference and its clinicopathologic significance of eight kinds of p53-related oncogenes and tumor-suppressor genes in HCC and adjacent-tumor liver tissues using tissue microarray technique.

METHODS

The HCC tissue microarrays comprised 273 cases of HCC tissues, 144 adjacent-tumor liver tissues, and 10 normal liver tissues obtained from autopsy. The diameter of each specimen on tissue microarrays was 2.0 mm. Immunohistochemistry was employed to detect the expression of p16(INK4a), p21(WAF1), p33(ING1b), p53, p57(KIP2), p73, mdm2, and ATM genes in tumor and adjacent tumor liver tissue, respectively. The relationship between HBV infection rate and the expression of these genes was also analyzed.

RESULTS

Three paraffin-embedded HCC tissue microarrays were successfully constructed, composed of 136, 143, and 148 tissue dots, respectively. The positive staining rates of these eight genes in HCC and surrounding-tumor tissues were: (p16(INK4a)) 35.9% and 9.0%, (p21(WAF1)) 41.8% and 11.1%, (p33(ING1b)) 43.65% and 14.6%, (p53) 46.2% and 12.5%, (p57(KIP2)) 39.2% and 16.7%, (p73) 9.5% and 2.8%, (mdm2) 41.4% and 9.0%, (ATM) 6.6% and 1.4%, respectively. The expression of all these genes in HCC were stronger than that in the surrounding-tumor liver tissues (P< 0.05). The expression differences of these genes in HCC tissues with varied differentiated grades were not significant(P >0.05). Except p16(INK4a), p21(WAF1), and p73 genes(P >0.05), the expression of p33(ING1b), p53, mdm2, ATM, and p57(KIP2) genes in HCC with portal vein invasion were higher than those in HCC without portal vein invasion (p33(ING1b), p53, mdm2, ATM, P< 0.01; p57(KIP2), P< 0.05). The infection rate of HBV did not have a significant correlation to the expression of these eight genes (P >0.05).

CONCLUSION

Tissue microarray technique had the advantage of high-throughput in the detection of HCC-related oncogenes and tumor-suppressor genes, which can analyze larger amounts of specimens, more target genes, and cost less working time.

摘要

背景与目的

肝癌发生是一个涉及多个基因的多阶段过程。分子生物学研究表明，p53基因家族的异常表达与肝细胞癌（HCC）的发生发展相关。本研究旨在利用组织芯片技术探讨8种p53相关癌基因和抑癌基因在HCC及癌旁肝组织中的表达差异及其临床病理意义。

方法

HCC组织芯片包含273例HCC组织、144例癌旁肝组织及10例尸检获得的正常肝组织。组织芯片上每个标本直径为2.0 mm。采用免疫组织化学方法分别检测肿瘤及癌旁肝组织中p16(INK4a)、p21(WAF1)、p33(ING1b)、p53、p57(KIP2)、p73、mdm2和ATM基因的表达。并分析HBV感染率与这些基因表达的关系。

结果

成功构建了3张石蜡包埋的HCC组织芯片，分别由136、143和148个组织点组成。这8种基因在HCC及癌旁组织中的阳性染色率分别为：(p16(INK4a)) 35.9%和9.0%，(p21(WAF1)) 41.8%和11.1%，(p33(ING1b)) 43.65%和14.6%，(p53) 46.2%和12.5%，(p57(KIP2)) 39.2%和16.7%，(p73) 9.5%和2.8%，(mdm2) 41.4%和9.0%，(ATM) 6.6%和1.4%。这些基因在HCC中的表达均强于癌旁肝组织（P<0.05）。这些基因在不同分化程度的HCC组织中的表达差异无统计学意义（P>0.05）。除p16(INK4a)、p21(WAF1)和p73基因外（P>0.05），有门静脉侵犯的HCC中p33(ING1b)、p53、mdm2、ATM和p57(KIP2)基因的表达高于无门静脉侵犯的HCC（p33(ING1b)、p53、mdm2、ATM，P<0.01；p57(KIP2)，P<0.05）。HBV感染率与这8种基因的表达无显著相关性（P>0.05）。