【作者】 丁敏；

【导师】 左国庆； 何松；

【作者基本信息】 重庆医科大学 ， 临床医学（专业学位）， 2019， 硕士

【摘要】 目的:探讨乙肝病毒X区基因(Hepatitis B virus X gene,HBX)突变情况与慢性乙肝病毒(hepatitis B virus,HBV)感染患者血清中血管内皮生长因子-C(VEGF-C)及血管内皮生长因子-D(VEGF-D)表达的相关性。方法:收集2018年4月至2018年10月于重庆医科大学附属第二医院就诊的HBV-DNA定量≥10^3拷贝/ml的慢性乙型病毒性肝炎患者血清55例,肝硬化50例,肝癌50例。提取患者血清中HBV-DNA,采用巢氏PCR方法扩增HBX基因,对产物进行基因测序,测序结果通过seqman软件与HBX基因野生型对比分析突变位点。酶联免疫吸附测定法检测患者血清中VEGF-C及VEGF-D浓度。通过卡方检验、单因素方差分析、秩和检验等方法对数据加以处理。结果:VEGF-C及VEGF-D血清浓度在肝癌组表达明显高于非肝癌组,差异具有统计学意义(P<0.05)。肝病患者血清中HBX常见的突变位点依次为:G1613A、A1652G、C1653T、T1753G、A1762T/G1764A、C1673T、G1676A,其中C1653T、A1762T/G1764A、T1753G突变率在肝癌组中明显高于非肝癌组,差异具有统计学意义(P<0.05)。采用秩和检验对二者进行分析发现,具有C1653T、A1762T/G1764A或T1753G三个位点突变的肝癌患者中VEGF-C的血清浓度显著高于不突变组,而VEGF-D的差异不具有统计学意义。结论:HBX中的C1653T、A1762T/G1764A和T1753G位点突变在肝癌患者中显著升高,且与VEGF-C的血清浓度相关,提示这三个突变位点可能在肝癌发生发展中发挥作用。更多还原

【Abstract】 Objective:To explore the relationship between Hepatitis B virus X gene(HBX)mutation and the serum expression of vascular endothelial growth factor-C(VEGF-C)and vascular endothelial growth factor-D(VEGF-D)in the patients with chronic hepatits B virus(HBV)infection.Methods:The serum samples of patients with HBV infection were collected in The Second Affiliated Hospital of Chongqing Medical University from April 2018 to October 2018,including 55 chronic hepatitis B patients,50 cirrhosis patients,and 50 HCC patients.The HBX gene sequences were amplificated from the serum samples by polymerase chain reaction(PCR).The amplified products were subsequently sequenced,and compared with those reported in GenBank to find the variable sites.The level of VEGF-C and VEGF-D from serum samples were detected by enzyme-linked immunosorbent assay.Results:The level of VEGF-C and VEGF-D were significantly higher in primary liver cancer group than other groups.The common HBX gene mutation sites include:G1613A,A1652 G,C1653T,T1753 G,A1762T/G1764 A,C1673T and G1676 A.The mutation rate of C1653 T,A1762T/G1764 A,T1753G were significantly higher in primary liver cancer group than other two groups.Furthermore,the three mutations are associated with higher level of VEGF-C in HCC patients by rank sum test.Conclusion : The C1653 T,A1762T/G1764 A and T1753 G site mutations in HBX are more frequent in HCC patients and associated with the expression of VEGF-C,implying that these mutations may be involved in the development and progression of hepatocellular carcinoma.更多还原