Article

Combined Antitumor Effect of Ad-bFGF-siRNA and Ad-Vpr on the Growth of Xenograft Glioma in Nude Mouse Model

Clinical Lab, Tianjin Huan Hu Hospital, Tianjin 300060, China.
Pathology & Oncology Research (Impact Factor: 1.56). 06/2011; 17(2):237-42. DOI: 10.1007/s12253-010-9303-5
Source: PubMed

ABSTRACT Basic fibroblast growth factor (bFGF) has been demonstrated to correlate with glioma grade and clinical outcome and has established its possible usefulness as a target for glioma therapy. Vpr has been described as an antitumor agent and displays a potent antitumor nature. Here, we try to investigate whether a combined treatment with bFGF-siRNA and Vpr gene would have a enhanced effectiveness on glioma in vitro and in vivo.After treatments with only Ad-bFGF-siRNA, only Ad-Vpr, and a combination of both, we assessed the changes in cell proliferation, cell cycle, and apoptosis in vitro by the methods of MTT, PI and FITC-AnnexinV double staining, respectively. In addition, we also evaluated the combined effect of bFGF-siRNA and Vpr gene therapy on glioma in vivo using xenograft glioma models in nude mice. Combined Ad-bFGF-siRNA and Ad-Vpr treatment was more better successful in inhibiting cell proliferation in comparison with treatments of either Ad-bFGF-siRNA or Ad-Vpr alone. Treatment of Ad-Vpr alone or a treatment of a combination of Ad-bFGF-siRNA and Ad-Vpr induced the G2/M cell cycle arrest and apoptosis; however, combined treatment was more effective than the Ad-Vpr treatment alone. Although each single treatment can slow the growth of xenograft glioma, the combined treatment with Ad-bFGF-siRNA and Ad-Vpr was better than either the Ad-bFGF-siRNA or Ad-Vpr treatment alone. Our results suggest that the combination therapy with bFGF-siRNA and Vpr gene can achieve a enhanced activity of anti-glioma, supporting the idea that the combination of these two antitumor agents could open new perspectives in glioma therapy.

0 Bookmarks
 · 
85 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: HIV-1 viral protein R (Vpr) inhibits cell growth and induces apoptosis in a wide range of cancers. However, the mechanism by which Vpr induces cell cycle arrest and apoptosis in GBM cell lines is unclear. The present work was taken to detect the proteins interacted with Vpr in U87MG cells. We analyzed the differential expression of proteins between glioblastoma cell U87MG treated with Ad-Vpr and untreated by 2-DE. We used antibody array analysis to analyze the common molecules in the apoptosis of U87MG induced by Vpr. We analyzed the differential expression of proteins between U87MG cell treated with Ad-Vpr and untreated, and found that proteins related to DNA damage repair or different apoptosis pathways were involved in the G2 arrest and apoptosis mediated by Vpr. In addition, proliferation-associated protein 2G4 (PA2G4), also known as Ebp1, was down-regulated and p53 was up-regulated in U87MG cells treated with Ad-Vpr. Our data suggest that Vpr may inhibit Ebp1 to stabilize p53, which in turn leads to G2 arrest and apoptosis in U87MG cells.
    Clinical and Translational Oncology 07/2013; DOI:10.1007/s12094-013-1072-7 · 1.28 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: OBJECTIVE: Studies have demonstrated that the transcription factor forkhead box P3 (FOXP3) is expressed not only in regulatory T cells, but also in some cancer cells. This study aims to clarify whether or not FOXP3 expression occurs in human gliomas and investigate the clinical significance of this expression in gliomas. METHODS: We detected FOXP3 protein expression in 40 glioma samples, 3 normal brain tissue samples, and 4 normal tonsil tissue samples using immunohistochemical staining and western blot. The expression of FOXP3 protein was also detected in five glioma cell lines by western blot. We also evaluated the association of FOXP3 expression with clinical pathological grades, prognosis, and recurrence. RESULTS: Western blot analysis showed that the expression of FOXP3 protein was upregulated in high-grade glioma (HGGS) samples compared with low-grade samples. The cell line U87 showed the highest FOXP3 expression, while U373 had the lowest expression. Immunohistochemical analysis detected FOXP3 protein in 35 out of the 40 (87.5 %) glioma samples and high levels of FOXP3 were observed in 26 out of the 27 (96.3 %) high-grade gliomas samples. Statistical analysis suggested that the upregulation of FOXP3 is significantly correlated with the histologic grade of gliomas (P < 0.05) and that patients with high expression of FOXP3 protein exhibit a poorer prognosis than those with low FOXP3 expression. CONCLUSIONS: Our findings suggest that FOXP3 expression in glioma cells has a crucial function in the development of HGGS and is associated with the malignant biological behavior of HGGS.
    Clinical and Translational Oncology 04/2013; 16(1). DOI:10.1007/s12094-013-1037-x · 1.28 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Colorectal cancer is a significant health problem, and the advanced stages of the disease have a low response rate to chemotherapy and easily acquire chemoresistance. HIV-1 viral protein R (Vpr) has been shown to possess inhibitory effects on various malignant cells in vivo and in vitro. In this study, an Ad-Vpr construct was used to infect the multidrug-resistant human colorectal cancer HCT-8/5-FU(MDR) cell line in vitro for cell viability, apoptosis, gene expression and gene activity using the MTT, flow cytometry, immunoblotting and gel shift assays, respectively. The data showed that Ad-Vpr significantly reduced HCT-8/5-FU(MDR) cell viability in a dose- and time-dependent manner. Ad-Vpr infection promoted HCT-8/5-FU(MDR) cells to undergo apoptosis and to arrest at the G2 phase of the cell cycle. The G2 cell cycle protein Cyclin B1 accumulated in the cells after Ad-Vpr infection. Furthermore, Ad-Vpr induced activation of caspase-3 and -9, but not caspase-8, in HCT-8/5-FU(MDR) cells. Ad-Vpr suppressed expression of the Bcl-xl protein, but upregulated Bax expression and cytochrome c release from the mitochondria in HCT-8/5-FU(MDR) cells. Ad-Vpr infection also resulted in a time-dependent decrease in nuclear translocation of NF-κB/p65 protein and p65 DNA-binding activity in HCT-8/5-FU(MDR) cells. The data from the current study provide mechanistic insights into understanding the molecular basis and utility of Ad-Vpr as a novel anticancer agent for multidrug resistance in human colorectal cancer.
    Oncology Reports 04/2012; 28(1):358-64. DOI:10.3892/or.2012.1782 · 2.19 Impact Factor