Medical Student Research Fellowship for Summer 2006

Mentor: Dr. Maria-Ana Ghetie and Dr. Ellen Vitetta
Department: Cancer Immunobiology Center
Room number: NB9.116
Mail Code: 8576
Phone number: 214-648-1221
E-mail: maria-ana.ghetie@utsouthwestern.edu,
ellen.vitetta@utsouthwestern.edu

Objective: to study the phenotype of a new multidrug resistant (MDR)
Human B-lymphoma cell line generated by drug selection

Project Type basic (translational) research

Brief Description of Project:
We have recently generated a new MDR human B-lymphoma cell line by drug selection in vitro (Raji resistant to Vincristine called Raji-VCR). We will now determine whether these cells express P-gp, CD20 and CD19. The aim of this project is to study the following characteristics of Raji-VCR in comparison with their parental counterpart (Raji) and with the well characterized Namalwa/MDR1 cells:
1. The expression of P-gp by direct or indirect FACS analysis.
2. The activity of the P-gp pump using the Rhodamine efflux assay.
3. The cytotoxicity of Vincristine using two methods: the inhibition of 3H-thymidine incorporation and CellTiter cell proliferation assay (a colorimetric method).
4. The distribution of P-gp between lipid rafts and soluble membrane fractions isolated in sucrose gradient from cells lysate using western blot analysis.
5. The quantitative distribution of proteins between the above fractions using a colorimetric method (BCA protein assay).
6. If time allows, the effect of at least one MAb (anti-CD20, Rituximab) on these new MDR cells could be investigated by the above methods.

Previous Research Activities or Publications with Medical Students: Only the students who worked with both Drs. Ghetie/Vitetta are included. There are 15 others not listed.

Medical Students who worked under Maria-Ana Ghetie
1) Mark Lowe - 1993
2) Vaught Elisabeth - 1995
3) Sami Arslanlar - 1996
4) Blake Sanders - 1997
5) Michelle Crank - 2002, 2003

Vitetta/Ghetie Publication with medical student:
Maria-Ana Ghetie, Michelle Crank, Stephanie Kufert, Iliodora Pop, and Ellen S. Vitetta: Rituximab but not other anti-CD20 antibodies reverses multidrug resistance in two B lymphoma cell line, blocks the activity of P-glycoprotein (P-gp), and induces P-gp to translocate out of lipid rafts (in press 2006, Journal of Immunotherapy).

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