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原发性肝癌患者PD-1基因拷贝数及其转录和蛋白表达水平的研究

ISSN号：1003-9279
期刊名称：《中华实验和临床病毒学杂志》
时间：0
分类：R512.62[医药卫生—临床医学;医药卫生—内科学]
作者机构：[1]广州市第八人民医院肝病科,510060
相关基金：广东省产业技术研究与开发资金计划项目（20078060401072）;广州市医药卫生科技重点项目（2007-ZDi-04）

关键词：肝肿瘤, 基因表达调控, 病毒, 细胞凋亡, Liver neoplasms, Gene expression regulation, viral, Apoposis

中文摘要：

目的检测原发性肝癌患者PD-1基因拷贝数及其转录和蛋白表达水平，研究宿主免疫调节因子PD-1基因拷贝数差异与肝癌的相关性。方法采用实时荧光定量PCR法检测24例原发性肝癌患者PD-1基因拷贝数和外周血单个核细胞PD-1 mRNA表达水平，采用流式细胞术检测外周血CD8＋T细胞PD-1蛋白表达水平。对照组为26例血清抗-HBs阳性健康者。结果肝癌组和对照组PD-1基因单拷贝率分别为34．62％和4．17％，多拷贝率分别为61．54％和95．83％，两组间分布差异有统计学意义（X^2=7．639，P=0．006），肝癌组人群PD-1基因多倍体率显著高于对照组。PD-1mRNA表达平均值对照组为2．35E-03，肝癌组为1．23E-03，Mann．whitey检验比较两组间表达差异有统计学意义，肝癌组明显低于健康对照组（U=153，P=0．009）；CD8＋T细胞PD-1蛋白表达频率对照组（3．72±0．32），肝癌组PD-1表达频率（16．13±1．68），肝癌组PD-1表达比健康对照组高，差异有统计学意义（t=-7．073，P=0．000）。结论PD-1基因多拷贝可能是HBV感染者发生肝癌的高危因素。PD-1基因拷贝数差异与原发性肝癌的关系值得进-步研究。

英文摘要：

Objective To study the gene copy number, mRNA transcription and protien expression of programmed cell death 1 （PD-1） gene in primary hepatocellular carcinoma （PHC） patients and normal control individuals （NC） who are anti-HBs positive, and to investigate the variations in PD-1 gene copy numbers and its relationship with PHC. Methods Real-time PCR was adopted to detect the PD-I gene copy numbers and their mRNA expressions in peripheral blood mononuclear cells （PBMCs） from 24 samples of PHC patients and 26 of NC. Protein expression level of PD-1 on CD8＋ T was analyzed by flow cytometry. Results In terms of number of PD-1 gene copy numbers, the percentage of cases of haploid （single） was 34. 62% and 4.17% in PHC group and control group respectively while the percentage of cases of diploid （double） was 61.54% and 95.83% respectively. The difference between the two was statistically significant （X2 = 7. 639 ,P = 0.006）. The rate of cases with double PD-I gene copy numbers was found to be higher in patients with PHC than in control group. It was also found that the average expression of PD-1 mRNA was 2. 35E-03 in control group and 1.23E-03 in PHC group. The expression level was significant lower in PHC group than that in control group when compared by using Mann-whitey technic （ U = 153, P = 0. 009 ）. Furthermore, the frequency of PD-1 protein expression on CD8＋ T cells was 3.72 ±0.32 in control group and 16.13 ± 1.68 in PHC group. The level of PD-1 mRNA expression was higher in PHC and significant differences was shown between two groups（ t = - 7. 073, P = 0. 000）. Conclusions Our study suggests that the variation in PD-1 gene copy number may trigger primary hepatocellular carcinoma to HBV carriers. The relationship between the variation of PD-1 gene copy numbers and its association with primary hepatocellular carcinoma is worth further focus.

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