title: TNF-α gene knockout in triple negative breast cancer cell line induces apoptosis.
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.
authors.affiliation: Faculty of Pharmacy, "Iuliu Hatieganu" University of Medicine and Pharmacy, 4 Pasteur Street, Cluj-Napoca 400349, Romania. valentinapilecki@gmail.com
authors.forename: Valentina
authors.initials: V
authors.lastname: Pileczki
authors.forename: Cornelia
authors.initials: C
authors.lastname: Braicu
authors.forename: Claudia D
authors.initials: CD
authors.lastname: Gherman
authors.forename: Ioana
authors.initials: I
authors.lastname: Berindan-Neagoe
mesh_terms: D017209:Apoptosis / Q000235:genetics*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D055786:Gene Knockout Techniques; D053263:Gene Regulatory Networks; D006801:Humans; D009336:Necrosis; D034741:RNA, Small Interfering / Q000378:metabolism; D013997:Time Factors; D064726:Triple Negative Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*
chemical_list: D034741:RNA, Small Interfering; D014409:Tumor Necrosis Factor-alpha