Guiqin Bai1, Fengping Fu, Yao Tang, Yueling Wang. 1. Department of Gynecology and Obstetrics, The First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an, China. baigq@126.com
Abstract
AIM: To investigate the effect and mechanism of hepatitis B virus (HBV) infection on the human choriocarcinoma cell line, JEG-3, in relation to apoptosis and intrauterine infection. MATERIAL AND METHODS: HBV-DNA serum was used to infect the choriocarcinoma cell line, JEG-3, in vitro. Real-time fluorescence quantitative PCR (RT-PCR) was then employed to detect intracellular replication of HBV DNA. Cells were also stained with Annexin-V and propidium iodide (PI) to identify the stages of apoptosis following infection. In addition, reverse transcription PCR was used to detect intracellular HBx mRNA levels, and Western blotting and immunohistochemistry were used to detect changes in the intracellular expression of HBxAg and phosphatidylinositol kinase 3 (PI3K). Flow cytometry was also used to detect the intracellular levels of phosphorylated AKT (pAKT). RESULTS: After JEG-3 cells were infected with HBV in vitro, HBV DNA was detected. The percentage of cells in early and late stage apoptosis also decreased significantly. Expression of HBx mRNA and HBxAg were detected, and intracellular levels of PI3K and pAKT were observed to significantly increase. CONCLUSION: HBV infected JEG-3 cells in vitro, resulting in an inhibition of early and late stage apoptosis. In addition, the HBxAg/PI3K/pAKT pathway is a possible mechanism mediating this inhibition of apoptosis, and the infection of the placenta by HBV.
AIM: To investigate the effect and mechanism of hepatitis B virus (HBV) infection on the humanchoriocarcinoma cell line, JEG-3, in relation to apoptosis and intrauterine infection. MATERIAL AND METHODS:HBV-DNA serum was used to infect the choriocarcinoma cell line, JEG-3, in vitro. Real-time fluorescence quantitative PCR (RT-PCR) was then employed to detect intracellular replication of HBV DNA. Cells were also stained with Annexin-V and propidium iodide (PI) to identify the stages of apoptosis following infection. In addition, reverse transcription PCR was used to detect intracellular HBx mRNA levels, and Western blotting and immunohistochemistry were used to detect changes in the intracellular expression of HBxAg and phosphatidylinositol kinase 3 (PI3K). Flow cytometry was also used to detect the intracellular levels of phosphorylated AKT (pAKT). RESULTS: After JEG-3 cells were infected with HBV in vitro, HBV DNA was detected. The percentage of cells in early and late stage apoptosis also decreased significantly. Expression of HBx mRNA and HBxAg were detected, and intracellular levels of PI3K and pAKT were observed to significantly increase. CONCLUSION:HBV infected JEG-3 cells in vitro, resulting in an inhibition of early and late stage apoptosis. In addition, the HBxAg/PI3K/pAKT pathway is a possible mechanism mediating this inhibition of apoptosis, and the infection of the placenta by HBV.