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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(t_1′=8.83、t_2′=6.59、t_3′=9.84,P<0.05);肝癌患者IFN-γ、IL-4水平明显低于健康者(t_1′= 6.67、t_2′=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。
Objective To investigate the variation of HBV gene, abnormal immune function and inactivation of tumor suppressor gene P16 in patients with hepatocellular carcinoma. Methods Using the gene chip and nucleotide sequence analysis technique, the serum HBV DNA (pre-C region 1814, 1896, BCP region 1762, 1764) in our hospital were analyzed. Flow cytometry was used to detect the peripheral blood P16 protein in WBC, T lymphocyte subsets by flow cytometry and IFN-γ, IL-4 levels by ELISA. Results The total HBV mutation rates in serum of 41 patients with chronic hepatitis B, 37 patients with hepatitis B cirrhosis and 36 patients with hepatocellular carcinoma were 58.5%, 75.6% and 88.8%, respectively. Liver cirrhosis , Respectively. The positive rates of P16 protein in peripheral blood of patients with chronic hepatitis B, hepatitis B cirrhosis and hepatocellular carcinoma were (43.09 ± 8.08)%, (30.54 ± 10.61)% and (16.29 ± 6.86)% and normal controls (76.57 ± 6.07)% respectively. The absolute numbers of CD3 +, CD4 +, CD8 + T lymphocytes in HCC patients were significantly lower than those in healthy controls (t 1 ’= 8.83, t 2’ = 6.59, t_3 ’= 9.84, P <0.05). The levels of IFN-γ and IL-4 in patients with liver cancer were significantly lower than those in healthy controls (t 1’ = 6.67, t 2 ’= 3.82, P <0.05). Conclusion HBV gene mutation is common in patients with liver cancer, with poor immune function and low expression of P16 protein in peripheral WBC.