SV40 T/t-Common Polypeptide Specifically Induces Apoptosis in Human Cancer Cells that Overexpress HER2/neu

Wen, Chun-Chiang; Cheng, Shih-An; Hsuen, Shu-Ping; Huang, Yaling; Kuo, Zong-Keng; Lee, Hsin-Fang; Kuo, Chou-Hua; Du, Jia-Ling; Wang, Won‐Bo

doi:10.1158/0008-5472.can-05-2109

Cited by 8 publications

(7 citation statements)

References 39 publications

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“…The lysates were incubated with the indicated antibody at 4°C for 18 h, and the protein G-agarose beads were then added and rotated at 4°C for 2 h. The beads were washed three times with lysis buffer, and the bound proteins were separated by SDS-PAGE followed by Western blotting with the indicated antibody. Western blot analyses were performed as described previously (56). Sub-G 1 apoptosis assays were performed as described previously (57).…”

Section: Methodsmentioning

confidence: 99%

Cellular Protein HAX1 Interacts with the Influenza A Virus PA Polymerase Subunit and Impedes Its Nuclear Translocation

Hsu

Shih

et al. 2013

J Virol

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Transcription and replication of the influenza A virus RNA genome occur in the nucleus through the viral RNA-dependent RNA polymerase consisting of PB1, PB2, and PA. Cellular factors that associate with the viral polymerase complex play important roles in these processes. To look for cellular factors that could associate with influenza A virus PA protein, we have carried out a yeast two-hybrid screen using a HeLa cell cDNA library. We identified six cellular proteins that may interact with PA. We focused our study on one of the new PA-interacting proteins, HAX1, a protein with antiapoptotic function. By using glutathione S-transferase pulldown and coimmunoprecipitation assays, we demonstrate that HAX1 specifically interacts with PA in vitro and in vivo and that HAX1 interacts with the nuclear localization signal domain of PA. Nuclear accumulation of PA was increased in HAX1-knockdown cells, and this phenotype could be reversed by reexpression of HAX1, indicating that HAX1 can impede nuclear transport of PA. As a consequence, knockdown of HAX1 resulted in a significant increase in virus yield and polymerase activity in a minigenome assay, and this phenotype could be reversed by reexpression of HAX1, indicating that HAX1 can inhibit influenza A virus propagation. Together, these results not only provide insight into the mechanism underlying nuclear transport of PA but also identify an intrinsic host factor that restricts influenza A virus infection.

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Section: Methodsmentioning

confidence: 99%

Cellular Protein HAX1 Interacts with the Influenza A Virus PA Polymerase Subunit and Impedes Its Nuclear Translocation

Hsu

Shih

et al. 2013

J Virol

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“…RHEK-1 cells (from Dr. Johng S. Rhim, Laboratory of Cellular and Molecular Biology, National Cancer Institute, USA), a nonmalignant cell line established from normal human foreskin keratinocytes infected with a hybrid virus adenovirus-12-simian virus-40 [39], RHEK-1 derivative cells (constructed in our lab; including RHEK/Tet-LMP1 [40], RHEK/Tet-On [40], LMP135 [41], RHEK-Vec, RHEK-endocan, RHEK-endocan-S21A, RHEK-endocan-F115,116A, and RHEK-endocan-S137A), NPC-TW04 cells, a human NPC cell line from Dr. Chin-Tarng Lin, Department of Pathology, College of Medicine, National Taiwan University [42], and H1299 cells, a human large cell lung carcinoma cell line from American Type Culture Collection (ATCC; Manassas, VA, USA), were cultured in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine and 1% penicillin/streptomycin. Primary human umbilical vein endothelial cells (HUVEC) were isolated from umbilical cord as described [43] and maintained in medium 199 (Invitrogen, Carlsbad, CA, USA) supplemented with 20% FBS, 30 μg/mL endothelial cell growth supplement (Upstate Biotechnology, Lake Placid, NY, USA), 15 µg/mL heparin (Leo Pharmaceutical Product, Ballerup, Denmark), and 1 mM pyruvate.…”

Section: Methodsmentioning

confidence: 99%

Upregulation of Endocan by Epstein-Barr Virus Latent Membrane Protein 1 and Its Clinical Significance in Nasopharyngeal Carcinoma

Chou

Chen³

et al. 2013

PLoS ONE

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Endocan (or called Esm-1) has been shown to have tumorigenic activities and its expression is associated with poor prognosis in various cancers. Latent membrane protein 1 (LMP1) is an Epstein-Barr virus (EBV)-encoded oncoprotein and has been shown to play an important role in the pathogenesis of EBV-associated nasopharyngeal carcinoma (NPC). To further understand the role of LMP1 in the pathogenesis of NPC, microarray analysis of LMP1-regulated genes in epithelial cells was performed. We found that endocan was one of the major cellular genes upregulated by LMP1. This induction of endocan by LMP1 was confirmed in several epithelial cell lines including an NPC cell line. Upregulation of endocan by LMP1 was found to be mediated through the CTAR1 and CTAR2 domains of LMP1 and through the LMP1-activated NF-κB, MEK-ERK and JNK signaling pathways. To study whether endocan was expressed in NPC and whether endocan expression was associated with LMP1 expression in NPC, the expression of endocan and LMP1 in tumor tissues from 42 NPC patients was evaluated by immunohistochemistry. Expression of endocan was found in 52% of NPC specimens. Significant correlation between LMP1 and endocan expression was observed (p<0.0001). Moreover, NPC patients with endocan expression were found to have a shorter survival than NPC patients without endocan expression (p=0.0104, log-rank test). Univariate and Multivariate analyses revealed that endocan was a potential prognostic factor for NPC. Finally, we demonstrated that endocan could stimulate the migration and invasion ability of endothelial cells and this activity of endocan was dependent on the glycan moiety and the phenylalanine-rich region of endocan. Together, these studies not only identify a new molecular marker that may predict the survival of NPC patients but also provide a new insight to the pathogenesis of NPC.

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“…rAd-T/t infection of SK-OV-3 cells could produce a significant amount of T/t-common protein, whereas rAd-V could not. 31 Moreover, rAd-T/t infection could inhibit HER2 expression and HER2 phosphorylation (that is, HER2 activity) in SK-OV-3 cells (Figure 1a). The infected cells were cultured in serum-free medium for 48 h and the CM was collected.…”

Section: Resultsmentioning

confidence: 94%

“…Previously, T/t-common has been shown to be able to induce apoptosis of HER2-overexpressing cancer cells under stressed conditions, such as low-serum condition. 31 It is possible that T/t-common, through inhibition of angiogenesis, creates a low-serumlike condition that may facilitate T/t-common to induce apoptosis in HER2-overexpressing tumors. Indeed, we found that rAd-T/t-treated HER2-overexpressing SK-OV-3 tumors contained fewer microvessels (Figure 6b) and underwent more severe apoptosis (Figure 6c) than rAd-V-treated SK-OV-3 tumors.…”

Section: Discussionmentioning

confidence: 99%

“…31 Preparation of concentrated conditioned medium In total, 3 Â 10 6 cells (SK-OV-3 or OVCAR-3) were infected with either rAd-V or rAd-T/t at appropriate multiplicity of infection, such that 490% of the cells were infected. The infected cells were incubated in medium containing 10% FBS for 24 h. The infected cells were then washed at least 10 times with phosphate-buffered saline (PBS) to remove uninfected viruses and incubated in serum-free medium.…”

Section: Adenoviral Constructsmentioning

confidence: 99%

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SV40 T/t-common polypeptide inhibits angiogenesis and growth of HER2-overexpressing human ovarian cancer

Cui

et al. 2011

Cancer Gene Ther

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Human epidermal growth factor receptor 2 (HER2) is frequently overexpressed in human ovarian cancers and its overexpression is associated with increased angiogenesis, increased metastasis and reduced survival. Inhibition of HER2 in HER2-overexpressing cancers can lead to reduced angiogenesis and improved survival. Previously, we reported that SV40 T/t-common polypeptide has transcriptional repression activity and can inhibit HER2 expression. In this study, we investigated the effect of T/t-common on the angiogenesis-inducing activity of HER2-overexpressing human SK-OV-3 ovarian cancer cells. We found that compared to conditioned medium from control SK-OV-3 cancer cells, conditioned medium from T/t-common-expressing SK-OV-3 cells had a reduced ability to induce endothelial cell migration and tube formation in vitro and microvessel formation in vivo. These data indicate that T/t-common can inhibit the ability of SK-OV-3 cancer cells to induce angiogenesis. T/t-common was found to be able to downregulate the expression of several proangiogenic factors, including vascular endothelial growth factor-A, interleukin-8, basic fibroblast growth factor, matrix metalloproteinase-2 and urokinase-type plasminogen activator, and upregulate antiangiogenic factors, including thrombospondin-1 and tissue inhibitor of metalloproteinases-1 in SK-OV-3 cancer cells. Finally, we demonstrated that T/t-common could inhibit the angiogenesis and growth of HER2-overexpressing human ovarian tumor in NOD/SCID mice. Taken together, the data suggest that T/t-common had the potential to be developed as a new antiangiogenic agent specific for treating HER2-overexpressing ovarian cancers.

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SV40 T/t-Common Polypeptide Specifically Induces Apoptosis in Human Cancer Cells that Overexpress HER2/neu

Cited by 8 publications

References 39 publications

Cellular Protein HAX1 Interacts with the Influenza A Virus PA Polymerase Subunit and Impedes Its Nuclear Translocation

Cellular Protein HAX1 Interacts with the Influenza A Virus PA Polymerase Subunit and Impedes Its Nuclear Translocation

Upregulation of Endocan by Epstein-Barr Virus Latent Membrane Protein 1 and Its Clinical Significance in Nasopharyngeal Carcinoma

SV40 T/t-common polypeptide inhibits angiogenesis and growth of HER2-overexpressing human ovarian cancer

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