5thANNUALNorthDakotaTribalCollegeResearchSymposium

Disseminating the Future of Tribal College Research

Friday,April 7, 2017

Sponsored by: NDIDeANetwork for Biomedical Research Excellence

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5thAnnual North Dakota Tribal College Research Symposium

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4thAnnual ND Tribal College Research Symposium

AbstractTemplate

An Investigation into the Differential Expressions of SCN1A and PFKFB3 Based onAbsence or Presence of the N- or C-Terminal Domains of MT-3

Derrick Fournier*, Brent J Voels, Scott H Garrett, Don A Sens, Seema Somji

Department of Pathology, School of Medicine and Health Sciences, 501 N. Columbia Road Stop 9037,University of North Dakota, Grand Forks, ND 58202-9037.

Studies have shown that the MT-3 protein contains 7 additional amino acids that are not present in any other member of the MT gene family, a 6 amino acid C-terminal sequence and a Thr in the N-terminal region. The goal of this study was to characterize the function of the N- and C-terminal domains of MT-3 in the breast cancer cell line, MCF-7. For this purpose six different constructs of MTs were prepared which were as follows: wild type (WT) MT-3, MT-3 N-terminal mutation, MT-3 C-terminal deletion, WT MT-1E, and MT-1E mutated to contain the N-terminal of MT-3 or the C-terminal or both the N- and the C-terminal of MT-3. Each of these constructs was stably transfected into MCF-7 cells and subsequent microarray analysis was performed. Microarray analysis demonstrated that Sodium Channel, Voltage Gated, Type 1 (SCN1A) and 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) expression were up regulated 2.35 and 2.1 fold respectively. It has been shown that voltage gated sodium channels (VGSCs) are up regulated in cancer, and favor an invasive/metastatic phenotype. Array validation demonstrated that SCN1A expression was not significantly altered. PFKFB3 is highly expressed and activated in cancer cells specifically colon adenocarcinoma. Real time PCR analysis indicated that PFKFB3 is significantly up regulated only when the N-terminal of MT-3 is present, but down regulated when only C-terminal is present. Differential regulation of the N- or C-terminal domains of MT-3 may affect tumor progression due to the alteration of PFKFB3 expression.