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The use of antisense oligonucleotides in evaluating survivin as a therapeutic target for radiation sensitization in lung cancer

Abstract

Elucidating the mechanism of over and under expression of proteins is critical in developing a better understanding of cancer. Multiple techniques are used to examine differential expression of proteins in cells and assess changes in protein expression in response to therapies such as radiation. Reduced expression can be caused by protein inactivation, mRNA instability, or reduced transcription. The following protocol was used to determine the mechanism for the reduced expression of an antiapoptotic factor, survivin, in normal tissues in response to radiation and the defect in cancer cells that prevents this reduction. We also examined ways to overcome survivin over expression in cancer cells in order to sensitize them to radiation. We will focus on the use of antisense oligonucleotides, cell cycle analysis, and luciferase reporter genes.

Abbreviations

HUVEC:

human umbilical vein endothelial cells

ASO:

antisense oligonucleotides

MDM2:

Murine Double Minute 2

PBS:

Phosphate Buffer Saline

IAP:

inhibitors of apoptosis family

7-AAD:

7-Aminoactinomycin D

DN:

Dominant Negative

MTT:

3-(4,5-methylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide

EBM-2:

endothelial basal medium-2

EGM-2:

endothelial growth media

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Correspondence to Bo Lu.

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Published: October 25, 2004.

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Shinohara, E.T., Hallahan, D.E. & Lu, B. The use of antisense oligonucleotides in evaluating survivin as a therapeutic target for radiation sensitization in lung cancer. Biol. Proced. Online 6, 250–256 (2004). https://doi.org/10.1251/bpo95

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  • DOI: https://doi.org/10.1251/bpo95

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