| Objectives To Provide the preliminary theory basis that HBx gene induces hepatic oval cells malignant transformation into hepatoma carcinoma cells, we investigated the effects of Hepatitis B virus X gene on the transdifferentiation of oval cells.Methods The changes of morphology, cell cycle, differentiated markers, oncogenes such as c-myc, TGF-É‘in pEGFP-HBx oval cells,which stably expressed HBx gene, were studied by Transmission Electron Microscopy, Flow cytometry, Periodic acid-Schiff (PAS) staining, Soft agar growth assay, Realtime-PCR, Immunocytochemistry. pEGFP-oval cells and LE/6 oval cells were used as control.Results (1) The pEGFP-HBx oval cells showed biger in size with malformed nucleus as compared with control. (2) Flow cytometry showed that ,in contrast with the control , the propotion of pEGFP-HBx oval cells arrested in G0/G1 phase decreased but in S or G2/M phase rose. Moreover, the population of aneuploid cells increased obviously. (3) Periodic acid-Schiff (PAS) staining showed many glycogen granules in the cytoplasm of pEGFP-HBx oval cells ; (4) The pEGFP-HBx oval cells formed colonies in the soft agar; (5) Compared with the control, the expression of HNF-4É‘, AFP, c-myc and TGF-É‘rose obviously, whereas the expression of CK-7 and CK-19 decreased.Conclusions The HBx gene can provoke abnormal differentiation of oval cells and induce oval cells malignant transformation. |
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