Chapter title |
Quantitative analysis of bortezomib-induced IL-8 gene expression in ovarian cancer cells.
|
---|---|
Chapter number | 27 |
Book title |
Cytokine Bioassays
|
Published in |
Methods in molecular biology, June 2014
|
DOI | 10.1007/978-1-4939-0928-5_27 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0927-8, 978-1-4939-0928-5
|
Authors |
Singha B, Phyo SA, Gatla HR, Vancurova I, Bipradeb Singha, Sai A. Phyo, Himavanth R. Gatla, Ivana Vancurova |
Abstract |
Interleukin-8 (IL-8), originally discovered as the neutrophil chemoattractant and inducer of leukocyte-mediated inflammation, contributes to cancer progression through its induction of tumor cell proliferation, survival, and migration. IL-8 expression is increased in many types of advanced cancers, including ovarian cancer, and correlates with poor prognosis. Bortezomib (BZ) is the first FDA-approved proteasome inhibitor that has shown remarkable antitumor activity in multiple myeloma and other hematological malignancies. In solid tumors, including ovarian carcinoma, BZ has been less effective as a single agent; however, the mechanisms remain unknown. We have recently shown that in ovarian cancer cells, BZ greatly increases IL-8 expression, while expression of other NFκB-regulated cytokines, IL-6 and TNF, is unchanged. In this chapter, we describe a protocol that uses real-time qRT-PCR to quantitatively analyze mRNA levels of IL-8 and IL-6 in BZ-treated ovarian cancer cells. The protocol can be easily modified and used for analysis of other cytokines in different cell types. |
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