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Quantitative analysis of bortezomib-induced IL-8 gene expression in ovarian cancer cells.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2014; Vol. 1172, pp. 295-304. - Publication Year :
- 2014
-
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.
- 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
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1172
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 24908316
- Full Text :
- https://doi.org/10.1007/978-1-4939-0928-5_27