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Names: Sarah Waliany and Shelina Kurwa

Names: Sarah Waliany and Shelina Kurwa High Schools: Flintridge Preparatory School, Westridge School for Girls Mentor: Dr. Susan Kane

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Names: Sarah Waliany and Shelina Kurwa

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  1. Names: Sarah Waliany and Shelina Kurwa • High Schools: Flintridge Preparatory School, Westridge School for Girls • Mentor: Dr. Susan Kane • Project Title: Transformation of Herceptin (Trastuzumab) Sensitive SKBR3 Breast Tumor Cells into Herceptin Resistant cells by Transfection with t-Darpp DNA Purpose: To show that t-Darpp DNA can induce Herceptin-resistance in previously sensitive Her-2 overexpressing SKBR3 breast cancer cells. Methodology: Two t-Darpp DNA fragments (3’ end and 5’ end) were combined to create a complete t-Darpp DNA strand by digestion, gel purification, ligation, and transformation. The complete t-Darpp DNA strand was transfected into Herceptin-sensitive Her-2 positive SKBR3 cells (from a company that sells established cell lines) without t-Darpp. These were then cultured with 0, 0.2 and 1.0 μM of Herceptin for 7, 14 and 21 days. An SRB Assay measured the protein biomass to determine cell growth. Western Blotting measured pAkt and Akt expression in the cells. Results: Exposure to Herceptin led to reduction in cell growth in cells without t-Darpp, whereas growth was much greater in the cells tranfected with t-Darpp. Unlike the control cells, the experimental cells expressed pAkt after Herceptin treatment. Conclusion: This is the first study that shows that Herceptin-sensitive breast cancer cells become resistant through transfection with t-Darpp. The clinically important result was that t-Darpp positive cells may proliferate more in the presence of Herceptin.

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