应用组织芯片检测肝细胞癌组织中8种p53相关基因的表达

背景及目的:肝细胞癌(hepatocellularcarcinoma,HCC)的发生是一个多步骤、多基因参与的过程。目前已知与HCC相关的癌基因和抑癌基因有十余种,其中p53基因家族的功能变化是HCC发生发展过程中的重要分子事件。本研究利用组织芯片高通量的优点,探讨肝细胞癌及癌旁肝组织中p53基因家族及相关基因中8种癌基因及抑癌基因的表达差异。方法:应用Instrumedics公司生产的组织芯片制作仪,取样针直径2mm,制作成3个肝细胞癌组织芯片蜡块,包括273例肝细胞癌组织、144例癌旁肝组织及10例非肝病死亡的尸检正常肝组织。应用免疫组织化学方法检测了p16INK4a、p21WAF1、p33ING1b、p53、p57KIP2、p73、mdm2和ATM8种基因在HCC及癌旁肝组织中的表达率,同时观察乙肝病毒(hapatitisBvirus,HBV)感染与这8种基因表达间的关系。结果:3个肝癌组织芯片分别含有136、143和148个位点。8种基因在HCC和癌旁肝组织中的阳性表达率分别为p16INK4a35.9%和9.0%、p21WAF141.8%和11.1%、p33ING1b43.65%和14.6%、p5346.2%和12.5%、p57KIP239.2%和16.7%、p739.5%和2.8%、mdm241.4%、9.0%和ATM6.6%和1.4%。在癌组织中上述基因的表达强于癌旁肝组织(P<0.05);不同分化程度的HCC组织中8种基因的阳性表达率差异均不显著(P>0.05);除了p16INK4a、p21WAF1、

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

BACKGROUND &OBJECTIVE:Hepatocarcinog enesis was a multistag e process involving a numb er of g enes.Molecular biolog ical st udies indicated that abnormal expression of p53g ene family is correlated with the development of hepatocellular carc inoma (HCC).This study was desig ned to investig ate the expression differe nce and its clinicopatholog ic sig ni ficance of eig ht kinds of p53-related oncog enes and tumor-suppressor g enes in HCC a nd adjacent-tumor liver tissues using tissue microarray technique.METHODS :The HCC tissue microarrays comprised 273cases of HCC tissues,144adjacent-tumor liver tissues ,and 10normal liver tissues obtained from autopsy.The diameter of each specimen on tiss ue microarrays was 2.0mm.Immunohistochemistry was employed to detect the expression of p16 INK4a ,p21 WAF1 ,p33 ING1b ,p53,p57 KIP2 ,p73,mdm2,and ATM g enes in tumor and adjacent tumor liver tissue,respectively.The relationship between HBV infection rate and the expression of these g enes was also analyzed.RESULTS :Three paraffin-embedded HCC tissue micro arrays were successfully construct ed,composed of 136,143,and 148tissue d ots,respectively.The positive sta ining rates of these eig ht g enes in HCC and s urrounding -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 g enes in HCC were strong er than that in the surrounding -tumor liver tissues(P<0.05).The expression differences of these g enes in HCC tissues with varie d differentiated g rades were not sig nificant (P>0.05).Except p16 INK4a ,p21 WAF1 ,and p73g enes (P>0.05),the expression of p33 ING1b ,p53,mdm2,ATM,and p57 KIP2 g enes in HCC with portal vein invasion were hig her 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 sig nificant correlation to the expression of the se eig ht g enes (P>0.05).CONCLUSION:Tissue microarray technique had the advantag e of hig h-throug hput in the detection of HCC-related oncog enes and tumor-suppressor g enes,which can a nalyze larg er amounts of specimens,more targ et g enes ,and cost less working t ime.