Publication Date

Spring 2016

Degree Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Advisor

Julio G. Soto

Keywords

downregulation of c-Myc, integrin subunit, recombinant mojastin, Sk-Mel-28, upregulation of p53 and TNF-B

Subject Areas

Molecular biology; Cellular biology

Abstract

Cell death by apoptosis is the result of one or multiple signaling pathways that can be initiated inside or outside the cell for a variety of reasons. Integrins, expressing themselves on the cell surface, play a central role in some apoptotic pathways. Cancer cells have been known to bypass this function and upregulate survival factor genes in order to elude apoptosis and thrive in the body, thus creating an environment in which it is possible for them to grow and metastasize. Previously in Dr. Soto’s lab, a recombinant disintegrin mutant, r-Moj-DM, was shown to induce apoptosis in melanoma cell line SK-Mel-28. To test our hypothesis that r-Moj-DM binds to the β1 integrin subunit, SK-Mel-28 cells were treated with a FITC-conjugated antibody anti-β1 and analyzed using flow cytometry. In order to ascertain which apoptotic pathways are activated by r-Moj-DM, ten genes, some known pro-apoptotic and some pro-survival, were investigated using RT-qPCR. It was confirmed that r-Moj-DM does bind to the β1 integrin subunit, and may induce apoptosis through that interaction. It was also found that two known pro-apoptotic genes, p53 and TNF-β, were upregulated in response to treatment with r-Moj-DM while c-Myc, a known survival factor, was downregulated. This shows the possibility that the intrinsic pathway may be activated due to a crossover from other pathways.

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