Jiangxi Provincial Key Laboratory of Molecular Medicine

Nanchang, China

Jiangxi Provincial Key Laboratory of Molecular Medicine

Nanchang, China

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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.


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.


PubMed | Jiangxi Provincial Key Laboratory of Molecular Medicine
Type: Journal Article | Journal: European journal of clinical investigation | Year: 2010

Fulminant hepatic failure (FHF) has a high mortality resulted from massive hepatic apoptosis and haemorrhage necrosis; it is required to develop a valid therapy directed towards hepatocyte protection and regeneration. Pim-3, a hepatic growth stimulator, belongs to the serine/threonine kinase Pim-family that has been implicated in gp130-mediated induction of cell proliferation, protection from apoptosis downstream of Signal transducer and activator of transcription 3 (STAT3) and vascular endothelial growth factor-A-dependent vasculogenesis and angiogenesis, thus is suggested to possibly play a role in the tissue repair of FHF.Male Wistar rats received simultaneous intraperitoneal injections of lipopolysaccharide (LPS) (100 microg kg(-1)) and D-galactosamine (D-GalN) (600 mg kg(-1)). One day prior to LPS/D-GalN administration, naked plasmid or Ringers solution was injected via tail vein by hydrodynamics-based procedure.Exogenous Pim-3 gene protected against LPS/D-GalN-induced lethality with survival rate of more than 80% and improved the hepatic pathomorphism. The fractions of hepatic apoptotic-positive cells and the levels of caspase-3 activity were markedly lower in Pim-3-pretreated rats. Furthermore, exogenous Pim-3 significantly inhibited expression of tumour necrosis factor-alpha and interleukin-1beta in the liver, declined p53 and inducible nitric oxide synthase mRNAs levels, but elevated levels of Bcl-2 protein, an anti-apoptosis member of Bcl-2 family, in the liver. Exogenous Pim-3, however, showed little effect on expression of Bax, a pro-apoptosis member of Bcl-2 family.Pim-3 gene could protect rats from FHF by inhibiting liver apoptosis and improving inflammatory response of liver tissues, which could be associated with inhibiting expression of inflammatory mediators and promoting expression of anti-apoptosis protein Bcl-2.

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