title: The effect of bovine rotavirus and its nonstructural protein 4 on ER stress-mediated apoptosis in HeLa and HT-29 cells.
abstract: Endoplasmic reticulum (ER) plays important roles in multiple cellular processes as well as cell survival and apoptosis. Perturbation of ER functions leads to ER stress and unfolded protein response (UPR). The primary goal of this response is cell survival, but severe ER stress can trigger apoptosis signaling. In tumor cells, chronically activated UPR response provides tumor growth. So, apoptosis induced by the ER stress has been the target for anti-cancer therapy. In this in vitro study, we examined the apoptotic effect associated with ER stress of bovine rotavirus and its nonstructural protein 4 (NSP4) alone in two cancer cell lines. The plasmid pcDNA3.1 encoding NSP4 protein of bovine rotavirus transfected with lipofectamine 2000 into the HeLa and HT-29 cells for protein production. MTT, flow cytometry, and Western blot were used to evaluate the cell viability, apoptosis, and expression level of C/EBP-homologous protein (CHOP) and activated caspase-4. In parallel, the apoptotic effect of the bovine rotavirus associated with ER stress in the infected cells was examined too. The cytotoxic and apoptotic effect of NSP4 protein on the cells were statistically significant compared to the control groups. However, Western blot showed that the expression of the NSP4 protein by recombinant plasmid did not lead to high expression of CHOP and activation of caspase-4. Interestingly, rotavirus not only induced significant apoptosis but also caused an increase in CHOP expression and caspase-4 activation in the infected cells compared to control. As a result, NSP4 protein and bovine rotavirus can be considered a potential novel bio-therapeutic strategy for cancer treatment.
authors.affiliation: Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Goodarzi
authors.affiliation: Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. soleim_h@modares.ac.ir.
authors.forename: Hoorieh
authors.initials: H
authors.lastname: Soleimanjahi
authors.affiliation: Department of Microbiology, School of Biology, College of Science, University of Tehran, Tehran, Iran.
authors.forename: Ehsan
authors.initials: E
authors.lastname: Arefian
authors.affiliation: Research and Development Department, Bayerpaul Group, Pardis Technology Park, Tehran, Iran.
authors.forename: Esmaeil
authors.initials: E
authors.lastname: Saberfar
mesh_terms: D000818:Animals; D017209:Apoptosis; D053471:Caspases, Initiator / Q000378:metabolism; D002417:Cattle; D059865:Endoplasmic Reticulum Stress / Q000502:physiology*; D006023:Glycoproteins / Q000235:genetics / Q000502:physiology*; D019073:HT29 Cells; D006367:HeLa Cells; D006801:Humans; D012401:Rotavirus / Q000502:physiology*; D014118:Toxins, Biological / Q000235:genetics / Q000502:physiology*; D051742:Transcription Factor CHOP / Q000032:analysis; D017361:Viral Nonstructural Proteins / Q000235:genetics / Q000502:physiology*
chemical_list: C497249:DDIT3 protein, human; D006023:Glycoproteins; C081369:NS28 protein, rotavirus; D014118:Toxins, Biological; D017361:Viral Nonstructural Proteins; D051742:Transcription Factor CHOP; C093516:CASP4 protein, human; D053471:Caspases, Initiator
keywords: Apoptosis; CHOP; Caspase-4; ER stress; Endoplasmic reticulum; Rotavirus