【摘要】 目的了解肝癌患者HBV基因变异、机体免疫功能异常及肿瘤抑制基因P16蛋白失活的情况。方法运用基因芯片和核苷酸序列分析技术,对114份我院住院患者血清中HBV DNA （前C区1814、1896、BCP区1762、1764位）进行分析,流式细胞荧光染色检测患者外周血WBC中P16蛋白,流式细胞仪检测T淋巴细胞亚群,ELISA法检测IFN-γ、IL-4水平。结果慢性乙型肝炎组（41份）、乙型肝炎肝硬化组（37份）、肝癌组（36份）血清中的HBV总突变率为58.5%、75.6%和88.8%,肝组织中肝硬化、肝癌分别为75.0%和100%;慢性乙型肝炎、乙型肝炎肝硬化、肝癌组患者外周血WBC中P16蛋白阳性率分别为（43.09±8.08）%、（30.54±10.61）%和（16.29±6.86）%,正常人为（76.57±6.07）%;肝癌患者CD3⁺、CD4⁺、CD8⁺T淋巴细胞绝对数均明显低于健康对照组（t₁′=8.83、t₂′=6.59、t₃′=9.84,P<0.05）;肝癌患者IFN-γ、IL-4水平明显低于健康者（t₁′= 6.67、t₂′=3.82,P<0.05）。结论肝癌患者中HBV基因变异普遍存在,机体免疫功能低下,外周血WBC中P16蛋白表达明显低。更多还原

【Abstract】 Objective To detect hepatitis B virus（HBV）genomic mutations,immune dysfunc- tion and inactivation of tumor suppressor gene P16 and their correlation with hepatocellular carcino- ma.Methods HBV gene mutation was detected by microarray and sequencing technology in serum of 114 cases.P16 proteins in peripheral blood leukocytes with fluorescence dyeing and T lymphocyte subsets were detected with flow cytometry.Serum interleukin-4（IL-4）and interferon gamma（IFN-γ） were detected by enzyme-linked immunosorbent assay（ELISA）.Results The gene mutation rate of HBV in serum was 58.5%,75.6% and 88.8% in 41 chronic hepatitis（CHB）and 37 liver cirrhosis （LC）and 36 hepatocellular carcinoma（HCC）cases,respectively,while the mutation rate of HBV in liver tissues was separately 75.0% and 100% in LC and HCC cases.The positive rate of P16 protein expressed in peripheral blood leukocytes of blood donors and patients with CHB,LC and HCC was （76.57±6.07）%,（43.09±8.08）%,（30.54±10.61）% and （16.29±6.86）%,respectively.The absolute numbers of CD3⁺ ,CD4⁺ and CD8⁺ T lymphocytes in patients with HCC were lower than those of normal adults（t₁′= 8.83,t₂′=6.59,t₃′=9.84,P<0.05）.The serum levels of IFN-γand IL-4 in patients with HCC were lower than those of normal aduhs（t₁′=6.67,t₂′= 3.82,P<0.05）. Conclusion HBV gene mutations are common,immune function and the expression of P16 are signif- icantly inhibited in the patients with hepatocellular carcinoma.更多还原