1. Quantitative analysis of bortezomib-induced IL-8 gene expression in ovarian cancer cells.
- Author
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Singha B, Phyo SA, Gatla HR, and Vancurova I
- Subjects
- Bortezomib, Cell Line, Tumor, Female, Humans, Interleukin-6 metabolism, Interleukin-8 metabolism, NF-kappa B genetics, NF-kappa B metabolism, RNA, Messenger isolation & purification, RNA, Messenger metabolism, Real-Time Polymerase Chain Reaction, Tumor Necrosis Factor-alpha genetics, Tumor Necrosis Factor-alpha metabolism, Antineoplastic Agents pharmacology, Boronic Acids pharmacology, Gene Expression drug effects, Interleukin-6 genetics, Interleukin-8 genetics, Pyrazines pharmacology, RNA, Messenger genetics
- 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.
- Published
- 2014
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