15-Deoxy-Δ12,14-prostaglandin J2(15d-PGJ2) Promotes Apoptosis Of HBx-positive Liver Cells

Objective: This study aims to investigate the inflammatory response characteristics ofliver cells caused by HBV x protein (HBx) including the changes of proliferation,apoptosis, and cycle of HL7702cell, and then study the unique impact of15d-PGJ2onHBx-positive liver cells.Methods: Liposome transfection and G418were used to to screen the transgenic cellwho stably express HBx. RT-PCR and Western blot were performed to detect theHBx mRNA and protein expression. Tetrazolium blue colorimetric method and flowcytometry were taken respectively to detect the proliferation, cycle, and apoptosis ofdifferent group of cells. Western blot were used to detect the apoptosis pathway-related proteins expression in each group of cells, such as Akt, NF-kB, Caspase-3,Bax and Bcl2.Results: HBx gene fragment and the HBx protein band can be amplified fromHL7702-HBx cells; The growth rate of HL7702after HBx gene transfection isobviously faster than that of HL7702cells(P<0.05); The flow cytometry test resultshows that the apoptosis rate of HL7702-HBx is significantly lower than that of theHL7702-NC cell. The cell proportion of HL7702-HBx in the G0/G1stage reducesgreatly, and the cell proportion in the S stage shows an increase. After the treatmentof15d-PGJ2, both the two cell lines proliferation are inhibited, and the inhibition rateswere increased with the inhibitors concentration; The apoptosis rates of HL7702-HBxand HL7702-NC increases after15d-PGJ2treatment, and the cell proportions in theG0/G1stage of the two groups of cells increased, whereas the cell proportions in the Sstage decrease. These changes are subject to the changes of HL7702-HBx cells. Thegreat changes also happen in the cell apoptosis pathway-related protein Akt, NF-kB,Caspase-3, Bax and Bcl2.Conclusion: HBx gene can promote the expression of Akt, NF-kB, the proteinconnecting with the apoptosis of cell receptor, which can be used to inhibit cell apoptosis. In the apoptosis pathway of the membrane receptor, HBx gene inbites theexpression of Bcl2but push the expression of Bax, which increases the cell apoptosis.It shows the two-way function of HBx on cell apoptosis. Under the function of drug15d-PGJ2,both HL7702-HBx cell and HL7702cell perform an increase in theexpression of promoting apoptosis protein, and a reduce in the expression ofinhibiting apoptosis protein. This change turns out to be more obvious in the changesof HL7702-HBx cells. Thus, a conclusion can be made that drug15d-PGJ2canpromote the cell apoptosis, whereas HBx gene might lead to a sensitive reaction ofliver cells to the apoptosis-promoting function of15d-PGJ2.