衰老标记蛋白-30的表达与肝疾病的关系

RELATIONSHIP BETWEEN EXPRESSION OF SENESCENCE MARKER PROTEIN-30 AND LIVER DISEASES

目的通过检测人正常肝组织、病毒性肝炎组织、肝硬化组织和肝癌(HCC)组织及相应癌旁组织中衰老标记蛋白-30(SMP-30)的表达,探讨SMP-30与肝疾病的关系。方法将本课题组构建的含SMP-30羧基端165个氨基酸的cDNA序列表达质粒,经表达和纯化获得MBP-SMP-30,免疫家兔制备出抗MBP-SMP-30多克隆抗体。采用免疫组织化学SABC法,检测30例人正常肝组织、10例病毒性肝炎组织、49例肝硬化组织和48例人HCC及相应癌旁组织中SMP-30的表达。结果SMP-30定位于肝细胞的细胞质和细胞核,其在HCC癌旁组织中的表达高于另外4种组织,具有显著性差异(P<0.05),且HCC中SMP-30的表达与肿瘤大小和病理分级无关。结论用MBP-SMP-30融合蛋白制备的多克隆抗体可成功检测出人肝细胞中的SMP-30。SMP-30与肝疾病的发生发展有关,其高表达可能是HCC的早期性、保护性事件。

Objective To detect the expression of SMP-30 protein in normal liver, virus hepatitis, hepatic cirrhosis, HCC and their corresponding paracancerous tissues, so as to study the relationship between SMP-30 and liver diseases. Methods The MBP-SMP-30 fusion protein was expressed and purified from expression plasmid that contained 165 amino acids of SMP-30 C- end. The fusion protein was used to immunize rabbit to prepare the anti-MBP-SMP-30 polyclonal antibody. Thirty normal liver, 10 virus hepatitis and 49 hepatic cirrhosis samples, 48 cases of HCC and their corresponding paracancerous tissues were used for detection. Immunohistochemical staining for SMP-30 was performed with SABC method on paraffin sections. Results SMP-30 was found in the cytoplasm and nucleus of hepatocyte. The protein expression level in HCC paracancerous tissues was higher than that in others （ P 〈 0.05）. The expression of SMP-30 protein in HCC was no correlation with tumor size and pathological grade. Conclusion SMP-30 in hepatocytes can be detected with polyclonal antibody of MBP-SMP-30 fusion protein. SMP-30 is associated with progression of liver diseases. The higher expression of SMP-30 protein may be an early and protective event in HCC.