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The Effect of Alpha3 Integrin on Neuroblastoma Cell Survival

CSEF · 2007 Biochemistry/ Molecular Biology

Jeffrey K. Lio · Torrey Pines High School, Encinitas, San Diego County

Overview

Objectives/Goals The objective is to determine whether the A isoform of the alpha3 integrin (alpha3 A) has an effect on short-term survival of neuroblastoma. Methods/Materials Alpha3 A was shuttled from the bacterial expression vector bluescript to the mammalian expression vector pcDNA3.1 (alpha3 A pcDNA3.1). NB5 was transfected with alpha3 A pcDNA3.1 and immunostained with anti-alpha3 antibody. NB& ov23 was transfected with alpha3 A pcNA3.1 and GFP (NB7 alpha3 A). Using a fluorescent microscope, the number of surviving NB7 alpha3 A in each window were recorded and images were taken daily. Results The NB5 alpha3 A pcDNA3.1 fluoresced after they were immunostained, whereas the immunostained NB5 transfected with empty vector pcDNA3.1 did not, indicating that neuroblastoma transfected with alpha3 A pcDNA3.1 did not express alpha3 integrin. NB7 ov23 cells transfected with alpha3 A pcDNA3.1 showed a decreased survival compared to that of NB7 ov23 transfected with empty vector pcDNA3.1 and the entire plate died after about one week. Conclusions/Discussion NB7 alpha3 A pcDNA3.1, which was confirmed to express alpha3 integrin, showed decreased survival compared to that of NB7 ov23 transfected with empty vector pcDNA3.1. Thus, the reinsertion of alpha3 A integrin into NB7 ov23, which is a cell line of NB7 that had lost its expression of alpha3 integrin, has a negative impact on the short-term survival. This suggests that the reinsertion of alpha3 integrin may inhibit tumorgenesis and metastasis in neuroblastoma.

Summary statement

Reinsertion of alpha3 integrin isoform A into neuroblastoma cells had a negative impact on short-term cell survival.

Help received

Lab equipment and materials at the UCSD Moores Cancer Center were provided by Dr. Dwayne Stupack, Associate Professor of Pathology. I worked in the lab under the supervision of Dr. Stupack and Dave Mikolon, the lab manager.

Competition history

  • CSEF 2007 Biochemistry/ Molecular Biology · Entry S0411

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