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TNF-α Gene Knockout in Triple Negative Breast Cancer Cell Line Induces Apoptosis
Faculty of Pharmacy, "Iuliu Hatieganu" University of Medicine and Pharmacy, 4 Pasteur Street, Cluj-Napoca 400349, Romania
Department of Functional Genomics and Experimental Pathology, Cancer Institute "Ioan Chiricută", 34–36 Republici Street, Cluj-Napoca 400015, Romania
Research Center for Functional Genomics, Biomedicine and Translational Medicine, "Iuliu Hatieganu" University of Medicine and Pharmacy, 8 Victor Babes Street, Cluj-Napoca 400012, Romania
Surgical Clinic II, 4–6 Clinicilor Street, Cluj-Napoca 400006, Romania
Department of Surgery, "Iuliu Haţieganu" University of Medicine and Pharmacy, 8 Victor Babes Street, Cluj-Napoca 400012, Romania
* Author to whom correspondence should be addressed.
Received: 9 October 2012; in revised form: 30 November 2012 / Accepted: 6 December 2012 / Published: 24 December 2012
Abstract: Tumor necrosis factor alpha (TNF-α) is a pro-inflammatory cytokine involved in the promotion and progression of cancer, including triple negative breast cancer cells. Thus, there is significant interest in understanding the molecular signaling pathways that connect TNF-α with the survival of tumor cells. In our experiments, we used as an in vitro model for triple negative breast cancer the cell line Hs578T. The purpose of this study is to determine the gene expression profiling of apoptotic signaling networks after blocking TNF-α formation by using specially designed siRNA molecules to target TNF-α messenger RNA. Knockdown of TNF-α gene was associated with cell proliferation inhibition and apoptosis, as observed by monitoring the cell index using the xCELLigence RTCA System and flow cytometry. PCR array technology was used to examine the transcript levels of 84 genes involved in apoptosis. 15 genes were found to be relevant after comparing the treated group with the untreated one of which 3 were down-regulated and 12 up-regulated. The down-regulated genes are all involved in cell survival, whereas the up-regulated ones are involved in and interact with pro-apoptotic pathways. The results described here indicate that the direct target of TNF-α in the Hs578T breast cancer cell line increases the level of certain pro-apoptotic factors that modulate different cellular networks that direct the cells towards death.
Keywords: RNA interference; apoptosis; cell signaling pathways; gene therapy
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Pileczki, V.; Braicu, C.; Gherman, C.D.; Berindan-Neagoe, I. TNF-α Gene Knockout in Triple Negative Breast Cancer Cell Line Induces Apoptosis. Int. J. Mol. Sci. 2013, 14, 411-420.
Pileczki V, Braicu C, Gherman CD, Berindan-Neagoe I. TNF-α Gene Knockout in Triple Negative Breast Cancer Cell Line Induces Apoptosis. International Journal of Molecular Sciences. 2013; 14(1):411-420.
Pileczki, Valentina; Braicu, Cornelia; Gherman, Claudia D.; Berindan-Neagoe, Ioana. 2013. "TNF-α Gene Knockout in Triple Negative Breast Cancer Cell Line Induces Apoptosis." Int. J. Mol. Sci. 14, no. 1: 411-420.