Date of Award

5-22-2019

Document Type

Master's Thesis - Restricted (NYMC/Touro only) Access

Degree Name

Master of Science

Department

Microbiology and Immunology

First Advisor

Dr. Raj K. Tiwari

Second Advisor

Dr. Jan Geliebter

Third Advisor

Dr. Xiumin Li

Abstract

Over the last three decades, melanoma has been diagnosed with an increased incidence. Melanoma is a cancer that arises from a wealth of mutations, including a BRAFV600E genetic lesion present in 50-60% of melanoma cases. The accumulation of mutations in melanoma makes it difficult to treat by conventional therapies and allows it to easily confer resistance to therapeutics. Despite the rise of novel immunotherapies that target melanoma, including small molecule inhibitors and checkpoint inhibitors, high side effects, low response rates, and acquired resistance have been observed. This bolsters the necessity of identifying alternative, supportive therapeutic strategies for potential combination therapies. Traditional Chinese Medicine (TCM) and Ayurveda, ancient systems of holistic medicine, have used formulations of compounds derived from plants and natural sources as treatment agents for thousands of years. Amongst these compounds are those that possess anti-inflammatory and anti-cancer properties. One such compound, Berberine (BBR), is a plant alkaloid isolated from Hydrastis canadensis (goldenseal), Coptis chinensis (goldenthread), Berberis aquifolium (Oregon grape), Berberis vulgaris (barberry), and Berberis aristate. Traditionally, it has been widely used for its antimicrobial and antiprotozoal properties. Current pre-clinical and clinical research on the impact of BBR in heart disease, diabetes, neurological disorders, aging, allergies, and many types of cancer is underway. We studied the effect of BBR on the proliferative abilities of three melanoma cells lines with different BRAF status: SKMEL28 (BRAFV600E+), SKMEL37 (BRAFV600E+/-), and SKMEL103 (BRAF wildtype). IC50 for cells was determined by XTT assay, which showed SKMEL103 has the lowest IC50 of 2.5 μL, making it the most susceptible to low concentration BBR treatment. Following this, cells were treated with their individual IC50 concentration (20-25 μL for BRAFV600E+ and 2.5 μL for BRAF wildtype) and effects on key cellular signaling pathways, including MAPK and PI3K/AKT/mTOR were studied by western blots. BBR treatment differentially downregulated phospho-MEK and phospho-ERK 1/2 dependent on presence of the BRAF genetic lesion. BBR treatment also downregulated expression of mTOR, which could consequentially result in regaining control of apoptosis and survival in cancer cells. BBR also arrested cell cycle progression in the G1 phase of the cell cycle after 72 hours of treatment in SKMEL37, making it a promising supportive therapeutic modality that may be used to supplement immunotherapies and small molecule inhibitors.

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