A novel real-time quantitative PCR method for analyzing HLA-C allele expression level

VernacularTitle:定量PCR检测HLA－C等位基因表达方法的建立和初步应用
Author: Wenjun WANG ; Sen LU ; Shichao WU ; Fengqin MIAO ; Ning PAN ; Jianqiong ZHANG
Publication Type:Journal Article
Keywords: Real time qPCR; HLA-C; Allelic specific; Liver cancer
From: Chinese Journal of Immunology 2016;32(8):1165-1170
CountryChina
Language:Chinese
Abstract: Objective: To establish real-time qPCR method to analyze each HLA-C allele expression level of individual.Methods:Database including exon 2&3 sequences of HLA class Ⅰalleles was built ,HLA-C allelic specific primers were designed and the real-time quantitative PCR method for analyzing HLA-C allele expression level was built .The allelic specificity of these primers were confirmed in database and 835 normal peripheral blood samples of Han population .The mRNA level of each HLA-C allele from 20 pairs of liver tumor tissues and non-tumor tissues was analyzed by the qPCR method we built .Results:20 pairs of allelic specific primers were designed to distinguish the two HLA-C alleles of each individual with frequency over 0.96%in 835 cases of Han population.Among 55%of the liver cancer tissues ,the expression levels of the two HLA-C alleles from the same tumor tissue changes differently compared to that of the relevant non-tumor tissue.Conclusion:This study provides method for HLA-C allele expression level analysis of Han population and each HLA-C allele expression level is inconsistent of liver cancer tissue .