Zhu Chengliang, Song Hui, Shen Bingzheng, Wu Long, Liu Fang, Liu Xinghui
Department of Clinical Laboratory, Renmin Hospital of Wuhan University, Wuhan, Hubei, 430060, People's Republic of China.
Department of Clinical Laboratory, Shanghai Gongli Hospital, the Second Military Medical University, Pudong New Area, Shanghai, 200135, People's Republic of China.
Lipids Health Dis. 2017 Jan 11;16(1):5. doi: 10.1186/s12944-016-0400-7.
Hepatitis B virus (HBV) infection causes acute and chronic liver disease, ultimately leading to the development of liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Phospholipase A2 group IIA (PLA2G2A) plays important roles in the development and progression of many tumors. Thus far, there have been no reports on the association between HBV and PLA2G2A. The present study investigated the effect of HBV infection on PLA2G2A expression and its application in the diagnosis of HBV-related diseases.
Serum levels of PLA2G2A in 308 HBV-infected patients and 185 healthy controls were measured using an enzyme-linked immunosorbent assay (ELISA). The difference in serum levels of PLA2G2A was analyzed among chronic hepatitis B (CHB), LC, and HCC patients. PLA2G2A mRNA and protein expression in HepG2 and HepG2.2.15 cells carrying the integrated HBV genome were measured using reverse transcription polymerase chain reaction (RT-PCR) and western blot assays. The HBV infectious clone pHBV1.3, the control plasmid pBlue-ks and PLA2G2A gene promoter were transfected into HepG2 and HepG2.2.15 cells. After transfection, the luciferase activity was measured in the cells. PLA2G2A mRNA and protein expression levels were examined using RT-PCR and western blot assays.
The serum levels of PLA2G2A were 258.3 ± 20.3ng/dl in the healthy controls and 329.0 ± 22.5ng/dl, 385.4 ± 29.3ng/dl and 459.2 ± 38.6ng/dl in the CHB, LC, and HCC patients, respectively. Statistical analyses revealed significantly higher serum levels of PLA2G2A in CHB, LC, and HCC patients than in the healthy controls (P < 0.05), and PLA2G2A levels were elevated in the order of HCC > LC > CHB group. High serum PLA2G2A levels in HCC patients were associated with a lower prevalence of lymph node metastasis and a lower TNM stage. HepG2.2.15 cells carrying the HBV genome expressed higher levels of PLA2G2A mRNA and protein than the HepG2 cells. In addition, HBV triggered PLA2G2A promoter activity and enhanced PLA2G2A mRNA and protein expression compared to the empty vector pBlue-ks.
HBV can upregulate the expression of PLA2G2A, and serum levels of PLA2G2A are associated with the progression of HBV-related diseases.
乙型肝炎病毒（HBV）感染可导致急慢性肝病，最终发展为肝硬化（LC）和肝细胞癌（HCC）。磷脂酶A2第IIA组（PLA2G2A）在许多肿瘤的发生和发展中起重要作用。迄今为止，尚无关于HBV与PLA2G2A之间关联的报道。本研究调查了HBV感染对PLA2G2A表达的影响及其在HBV相关疾病诊断中的应用。
采用酶联免疫吸附测定（ELISA）检测308例HBV感染患者和185例健康对照者血清中PLA2G2A水平。分析慢性乙型肝炎（CHB）、LC和HCC患者血清PLA2G2A水平的差异。采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测携带整合型HBV基因组的HepG2和HepG2.2.15细胞中PLA2G2A mRNA和蛋白表达。将HBV感染性克隆pHBV1.3、对照质粒pBlue-ks和PLA2G2A基因启动子转染至HepG2和HepG2.2.15细胞。转染后，检测细胞中的荧光素酶活性。采用RT-PCR和蛋白质免疫印迹法检测PLA2G2A mRNA和蛋白表达水平。
健康对照者血清PLA2G2A水平为258.3±20.3ng/dl，CHB、LC和HCC患者血清PLA2G2A水平分别为329.0±22.5ng/dl、385.4±29.3ng/dl和459.2±38.6ng/dl。统计学分析显示，CHB、LC和HCC患者血清PLA2G2A水平显著高于健康对照者（P<0.05），且PLA2G2A水平按HCC>LC>CHB组顺序升高。HCC患者血清PLA2G2A高水平与较低的淋巴结转移率和较低的TNM分期相关。携带HBV基因组的HepG2.2.15细胞比HepG2细胞表达更高水平的PLA2G2A mRNA和蛋白。此外，与空载体pBlue-ks相比，HBV激活PLA2G2A启动子活性并增强PLA2G2A mRNA和蛋白表达。
HBV可上调PLA2G2A表达，血清PLA2G2A水平与HBV相关疾病的进展有关。
