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Ding H.,Nanchang University | Ding H.,Jiangxi Provincial Key Laboratory of Molecular Medicine | Xu J.-J.,Jiangxi Provincial Key Laboratory of Molecular Medicine | Huang Y.,Nanchang University | And 3 more authors.
Biochemical and Biophysical Research Communications | Year: 2012

Objectives: We investigated the effects of xeroderma pigmentosum D (XPD) on the growth of hepatoma cells and the expressions of P21, Bax, Bcl-2 and Hepatitis B virus X protein (HBx). In addition, we examined whether XPD affected the aforementioned genes via the P53 pathway. Methods: Human hepatoma cells (HepG2.2.15) were transfected with XPD expression vector, followed by incubation with Pifithrin-α (P53 inhibitor). By using RT-PCR and Western blotting, the expression levels of XPD, P53, phospho-P53 (ser-15), P21, Bax, Bcl-2 and HBx were detected. The cell cycle and the apoptosis rate were examined with flow cytometry, and the cell viability was detected by MTT. Results: Over-expression of XPD up-regulated the expressions of P53, phospho-P53 (ser-15), P21 and Bax but down-regulated the expressions of Bcl-2 and HBx. XPD inhibited the viability of HepG2.2.15 and exacerbated the apoptosis. However, the inhibition of P53 by Pifithrin-α abolished the above-mentioned effects of XPD. Conclusion: XPD could suppress growth of hepatoma cells, up-regulate the expressions of P21 and Bax, and down-regulate the expressions of Bcl-2 and HBx through the P53 pathway. There may be mutual influences among XPD, P53 and HBx that co-regulate hepatocarcinogenesis. © 2012 Elsevier Inc.

Yan C.,Nanchang University | Yan C.,Jiangxi Provincial Key Laboratory of Molecular Medicine | Liu X.-X.,Jiangxi Provincial Key Laboratory of Molecular Medicine | Ge J.,Nanchang University | And 5 more authors.
Tumor | Year: 2014

Objective: To investigate the effects of RNA interference targeting eukaryotic translation elongation factor 1A1(eEF1A1) expression on the proliferation of hepatocellular carcinoma Hep3B cells, and to explore its possible mechanism. Methods: A recombinant vector pGPU6/GFP/Neo-eEF1A1-shRNA containing short hairpin RNA (shRNA) targeting eEF1A1 was constructed, and then it was transfected into the Hep3B cells. After tansfection with pGPU6/GFP/Neo-eEF1A1-shRNA, the expressions of eEF1A1mRNA and protein were examined by real time fluorescence quantitative PCR and Western blotting, respectively, the cellular growth ability was examined by cell counting kit 8 (CCK-8) assay, the colony formation ability was detected by colony formation assay, and the cell cycle distribution of Hep3B cells was detected by flow cytometry (FCM). The expression levels of cyclin D1 and cyclin-dependent kinase 4 (CDK4) proteins in Hep3B cells were examined by Western blotting. Results: The recombinant vector pGPU6/GFP/Neo-eEF1A1-shRNA was successfully constructed, and the Hep3B cells with stable expession of eEF1A1 were established. The expression levels of eEF1A1mRNA and protein in Hep3B cells after transfection with pGPU6/GFP/NeoeEF1A1-shRNA were lower than those in the negative control cells (Hep3B cells transfected with a negative control vector pGPU6/GFP/Neo-NC) and the blank control cells (Hep3B cells without any transfection) (P < 0.01, P < 0.05). As compared with the negative control, the cellular growth ability and the colony formation ability of Hep3B cells after tranfection with pGPU6/GFP/Neo-eEF1A1-shRNA were obviously decreased (P < 0.05, P < 0.01); the percentage of the cells in G1 phase was increased (P < 0.01), and which in S-phase was decreased (P < 0.05); the expression levels of cyclin D1and CDK4 proteins were down-regulated (both P < 0.05). Conclusion: Down-regulation of eEF1A1 expression can inhibit the proliferation of Hep3B cells. Copyright © 2014 by TUMOR.

Wang H.-Y.,Nanchang University | Wang H.-Y.,Jiangxi Provincial Key Laboratory of Molecular Medicine | Xiong G.-F.,Nanchang University | Xiong G.-F.,Jiangxi Provincial Key Laboratory of Molecular Medicine | And 7 more authors.
Medical Oncology | Year: 2012

We investigated the role of XPD in cell apoptosis of hepatoma and its relationship with p53 during the regulation of hepatoma bio-behavior. RT-PCR and Western blot were used to detect the expression levels of XPD, p53, c-myc, and cdk2. The cell apoptosis and cell cycle were analyzed with flow cytometry. Compared with the control cells, XPD-transfected cells displayed a lower viability and higher apoptosis rate. A decreased expression of p53 gene was detected in XPD-transfected cells. In contrast, both c-myc and cdk2 showed increased expressions of mRNAs and proteins in the transfected cells. Our results indicate that XPD may play an important role in cell apoptosis of hepatoma by inducing an over-expression of p53, but suppressing expressions of c-myc and cdk2. © 2011 Springer Science+Business Media, LLC.

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