Interleukin-8 (IL-8), originally discovered as the neutrophil chemoattractant and inducer of leukocytemediated 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.
CITATION STYLE
Singha, B., Phyo, S. A., Gatla, H. R., & Vancurova, I. (2014). Quantitative analysis of Bortezomib-Induced IL-8 gene expression in ovarian cancer cells. Methods in Molecular Biology, 1172, 295–304. https://doi.org/10.1007/978-1-4939-0928-5_27
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