Study On The Correlation Between PI4KA And Hepatitis B Virus Replication

Objective:To investigate the relationship between serum Phosphatidylinositol4-kinase Ⅲ alpha (PI4KA) and HBV DNA loads in patients with chronic hepatitis B; To study the expression of PI4KA) in hepatoma cell lines HepG2and HepG2.2.15,and explore the effect of hepatitis B virus (HBV) on the expression of PI4KA. To investigated the relationship between PI4KA and HBV replication both at the levels of clinical and cells.Methods:Serum samples of180patients with chronic hepatitis B were collected and the HBV DNA loads were tested by fluorescence quantitative PCR. According to the serum HBV DNA loads, patients were divided into three groups:A high-load group with DNA loads higher than107copies/ml, a medium-load group with DNA loads between105copies/ml and107copies/ml, and a low-load group with DNA loads lower than105copies/ml. Serum samples of60healthy people were included as control. The serum levels of PI4KA in each group were quantified by ELISA. The hepatoma cell lines HepG2and HepG2.2.15were well cultured, when the two hepatocytes grew to90%, the RNA of the two hepatocytes were extracted and then were reversely transcribed to cDNA;the expressions of PI4KA mRNA in two hepatocytes were determined by quantitative real-time RT-PCR; the levels of PI4KA in cell culture medium were quantified by ELISA method.Results:The level of serum PI4KA was2.29±0.75ng/ml in the60healthy control,2.73±0.71ng/ml in the60patients with a low-load group,3.52±0.78ng/ml in the60patients with a medium-load group and4.72±0.77ng/ml in the60patients with a high-load group, respectively. The levels of serum PI4KA showed statistically significant differences among the groups with different viral loads and healthy control (F=119.958, P<0.01), and PI4KA expression in groups with different viral loads was higher than that of the healthy control. Serum PI4KA expression was higher with the increasing of the HBV DNA loads, and there was a positive correlation between the levels of serum PI4KA and the serum HBVDNA loads (r=0.758, P＜0.01). The expression of PI4KA mRNA in HepG2.2.15cells were high than that in HepG2cells and the difference was statistically significant (P＜0.05); the levels of PI4KA in HepG2.2.15cell culture medium were significantly higher than the HepG2cells (1.63±0.43vs1.07±0.54;t=2.29, P＜0.05).Conclusion:The levels of Serum PI4KA display a closely relationship with the HBV DNA loads; the expression of PI4KA in HepG2.2.15cell are up-regulated on the levels of both mRNA and proteins. We demonstrate that PI4KA display a closely relationship with HBV replication both at clinical and cell levels. HBV may up-regulate the expression of PI4KA to benefit its survival and replication.