<i>In-vitro</i>Characterization and Evaluation of Apoptotic Potential of Bicistronic Plasmid Encoding HN Gene of Newcastle Disease Virus and Human TNF-α

Rajmani, R. S.; Singh, Prafull; Kumar, Girish; Saxena, Shikha; Singh, L. P.; Kumar, Rajiv; Sahoo, Aditya Prasad; Gupta, Shishir Kumar; Chaturvedi, Uttara; Tiwari, Ashok Kumar

doi:10.1080/10495398.2014.933741

Cited by 12 publications

(10 citation statements)

References 21 publications

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“…TNF-α has been recognized as an important host defense cytokine that affects tumor cells. One of the most prominent characteristics of TNF-α is that it is capable of functioning together with endothelial cells to cause apoptosis of tumor, resulting in tumor necrosis (Rajmani et al, 2015). The present results showed that the serum levels of IL-2, IL-6, TNF-α and IFN-γ were slightly restored and enhanced after PD treatment in a dose-dependent manner compared with those in the vehicle control group in H22 tumor-bearing mice.…”

Section: Discussionmentioning

confidence: 44%

Platycodin D exerts anti-tumor efficacy in H22 tumor-bearing mice via improving immune function and inducing apoptosis

Tian

Liu

et al. 2016

J. Toxicol. Sci.

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-Platycodin D (PD), a major saponin derived and isolated from the roots of Platycodon grandiflorum, exerts potent growth inhibition and strong cytotoxicity against various cancer cell lines. However, the anti-tumor efficacy of PD on H22 hepatocellular carcinoma remains unknown. In the present study, we aimed to explore the anti-hepatoma activity in vivo and the underlying mechanism of PD in H22 tumor-bearing mice. The results revealed that PD could considerably suppress tumor growth with no significant side effects on immune organs and body weight. Further investigations showed that the levels of serum cytokines, including interferon gamma (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-2 (IL-2), were enhanced by PD administration. On the other hand, PD inhibited the production of vascular endothelial growth factor (VEGF) in serum of H22 tumor mice. Additionally, the observations from H&E and Hoechst 33258 staining results demonstrated that PD noticeably induced apoptosis in H22 hepatocellular carcinoma cells. Importantly, immunohistochemical analysis showed that PD treatment increased Bax expression and decreased Bcl-2 and VEGF expression of H22 tumor tissues in a dose-dependent manner. Taken together, the findings in the present investigation clearly demonstrated that the PD markedly suppressed the tumor growth of H22 transplanted tumor in vivo at least partly via improving the immune functions, inducing apoptosis, and inhibiting angiogenesis.

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Section: Discussionmentioning

confidence: 44%

Platycodin D exerts anti-tumor efficacy in H22 tumor-bearing mice via improving immune function and inducing apoptosis

Tian

Liu

et al. 2016

J. Toxicol. Sci.

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“…In our previous studies, we showed that the HN protein can cause apoptosis in chicken embryo fibroblasts and in HeLa cells [14,21]. We have demonstrated the activation of the SAPK/JNK pathway by HN [13].…”

Section: Discussionmentioning

confidence: 83%

“…HeLa cells are naturally resistant to TNF-a-induced cytotoxicity but may become sensitized when NF-jB transactivation is blocked or cells are treated with compounds that inhibit protein synthesis [20]. We proposed in the previous study that HN may also sensitize otherwise resistant HeLa cells to TNF-a by exerting an inhibitory effect on NF-jB activation through the accumulation of IFN-a [21]. With this perspective in mind, we assessed if HN protein could sensitize HeLa cells to TNF-a-induced cell death and if a combination of HN and TNF-a could exert a synergistic apoptotic effect.…”

Section: Discussionmentioning

confidence: 99%

“…Cloning and expression of the HN gene of NDV (pVIVO.nd.hn), human TNF-a (pVIVO.hu.tnf) and a bicistronic clone of HN ? TNF-a (pVIVO.nd.hn.hu.tnf) in pVIVO2-mcs were done previously in our laboratory [21]. Recombinants were used to transfect 70-80 % confluent HeLa cells in a 24-well plate using Lipofectamine TM transfection reagent as per the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

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HN protein of Newcastle disease virus sensitizes HeLa cells to TNF-α-induced apoptosis by downregulating NF-κB expression

et al. 2016

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Newcastle disease virus (NDV) is an oncolytic virus which selectively replicates in tumor cells and exerts anti-tumor cytotoxic activity by promoting cell death. In this study, we focus on characterization of the underlying mechanisms of NDV-induced cell death in HeLa cells. We find that NDV Herts/33 strain triggers both extrinsic and intrinsic apoptosis at late infection times. The activation of NF-кB pathway and subsequent up-regulation of TNF-α/TRAIL initiates extrinsic apoptosis, leading to activation of caspase 8 and cleavage of Bid into tBid. tBid transmits the extrinsic apoptotic signals to mitochondria and mediates intrinsic apoptosis, which is hallmarked by cleavage of caspase 9. Moreover, RIP1 is cleaved into RIP1-N and RIP1-C at D324 by caspase 8, and this cleavage promotes apoptosis. Surprisingly, over expression of RIP1 reduces apoptosis and depletion of RIP1 promotes apoptosis, suggesting full length RIP1 is anti-apoptotic. Moreover, necroptosis hallmark protein MLKL is activated by phosphorylation at 12-24 h.p.i., and RIP1 regulates the level of phosphor-MLKL. Immunostaining shows that RIP1 aggregates to stress granules (SGs) at 8-24 h.p.i., and phosphor-MLKL is also recruited to SGs, instead of migrating to plasma membrane to exert its necrotic function. Immunoprecipitation study demonstrates that RIP1 bind to phosphor-MLKL, and depletion of RIP1 reduces the aggregation of MLKL to SGs, suggesting that RIP1 recruits MLKL to SGs. Altogether, NDV infection initiates extrinsic apoptosis via activation of NF-кB and secretion of TNF-α/TRAIL. Activation of caspase 8 by TNF-α/TRAIL and subsequent cleavage of Bid and RIP1 transmit the death signals to mitochondria. Meanwhile, virus subverts the host defensive necroptosis via recruiting phosphor-MLKL by RIP1 to SGs. Thus, RIP1 is a central signaling protein in regulation of apoptosis and necroptosis during NDV infection. www.impactjournals.com/oncotarget/

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“…It was previously shown that the HN protein plays a critical role in the anti-tumor effects of NDV (10). HN hydrolyzes the surface sialic acid of the host cell, exposes biological recognition sites, and induces tumor necrosis factor-associated apoptosis-inducing ligand (TRAIL) expression at the surface of mononuclear cells in the host peripheral blood, and yet does so independently of viral replication (11,12). In addition, HN positioning in the tumor cell membrane also led to formation of identical recognition sites, which in turn improved the cytotoxic effects of the host immune system against tumor cells (13).…”

Section: Introductionmentioning

confidence: 99%

Therapeutic targeting of liver cancer with a recombinant DNA vaccine containing the hemagglutinin-neuraminidase gene of Newcastle disease virus via apoptotic-dependent pathways

et al. 2016

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A total of ~38.6 million mortalities occur due to liver cancer annually, worldwide. Although a variety of therapeutic methods are available, the efficacy of treatment at present is extremely limited due to an increased risk of malignancy and inherently poor prognosis of liver cancer. Gene therapy is considered a promising option, and has shown notable potential for the comprehensive therapy of liver cancer, in keeping with advances that have been made in the development of cancer molecular biology. The present study aimed to investigate the synergistic effects of the abilities of the hemagglutinin neuraminidase protein of Newcastle disease virus (NDV), the pro-apoptotic factor apoptin from chicken anaemia virus, and the interferon-γ inducer interleukin-18 (IL-18) in antagonizing liver cancer. Therefore, a recombinant DNA plasmid expressing the three exogenous genes, VP3, IL-18 and hemagglutinin neuraminidase (HN), was constructed. Flow cytometry, acridine orange/ethidium bromide staining and analysis of caspase-3 activity were performed in H22 cell lines transfected with the recombinant DNA plasmid. In addition, 6-week-old C57BL/6 mice were used to establish a H22 hepatoma-bearing mouse model. Mice tumor tissue was analyzed by immunohistochemistry and scanning electron microscopy. The results of the present study revealed that the recombinant DNA vaccine containing the VP3, IL-18 and HN genes inhibited cell proliferation and induced autophagy via the mitochondrial pathway in vivo and in vitro.

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In-vitroCharacterization and Evaluation of Apoptotic Potential of Bicistronic Plasmid Encoding HN Gene of Newcastle Disease Virus and Human TNF-α

Cited by 12 publications

References 21 publications

Platycodin D exerts anti-tumor efficacy in H22 tumor-bearing mice via improving immune function and inducing apoptosis

Platycodin D exerts anti-tumor efficacy in H22 tumor-bearing mice via improving immune function and inducing apoptosis

HN protein of Newcastle disease virus sensitizes HeLa cells to TNF-α-induced apoptosis by downregulating NF-κB expression

Therapeutic targeting of liver cancer with a recombinant DNA vaccine containing the hemagglutinin-neuraminidase gene of Newcastle disease virus via apoptotic-dependent pathways

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