Interleukin-29 binds to melanoma cells inducing Jak-STAT signal transduction and apoptosis.
Interleukin-29 (IL-29) is a member of the type III IFN family that has been shown to have antiviral activity and to inhibit cell growth. Melanoma cell lines were tested for expression of the IL-29 receptor (IL-29R) and their response to IL-29. Expression of IL-28R1 and IL-10R2, components of IL-29R, was evaluated using reverse transcription-PCR. A combination of immunoblot analysis and flow cytometry was used to evaluate IL-29-induced signal transduction. U133 Plus 2.0 Arrays and real-time PCR were used to evaluate gene expression. Apoptosis was measured using Annexin V/propridium iodide staining. In situ PCR for IL-29R was done on paraffin-embedded melanoma tumors. Both IL-28R1 and IL-10R2 were expressed on the A375, 1106 MEL, Hs294T, 18105 MEL, MEL 39, SK MEL 5, and F01 cell lines. Incubation of melanoma cell lines with IL-29 (10-1,000 ng/mL) led to phosphorylation of signal transducer and activator of transcription 1 (STAT1) and STAT2. Microarray analysis and quantitative reverse transcription-PCR showed a marked increase in transcripts of IFN-regulated genes after treatment with IL-29. In the F01 cell line, bortezomib-induced and temozolomide-induced apoptosis was synergistically enhanced following the addition of IL-29. In situ PCR revealed that IL-10R2 and IL-28R1 were present in six of eight primary human melanoma tumors but not in benign nevi specimens. In conclusion, IL-29 receptors are expressed on the surface of human melanoma cell lines and patient samples, and treatment of these cell lines with IL-29 leads to signaling via the Jak-STAT pathway, the transcription of a unique set of genes, and apoptosis.
Guenterberg
Kristan D
KD
Department of Surgery, Division of Surgical Oncology, The Ohio State University, Arthur G James Cancer Hospital and Richard J SoloveResearch Institute, Columbus, Ohio 43210, USA.
Grignol
Valerie P
VP
Raig
Ene T
ET
Zimmerer
Jason M
JM
Chan
Anthony N
AN
Blaskovits
Farriss M
FM
Young
Gregory S
GS
Nuovo
Gerard J
GJ
Mundy
Bethany L
BL
Lesinski
Gregory B
GB
Carson
William E
WE
D017209:Apoptosis; D001897:Boronic Acids / Q000494:pharmacology; D000069286:Bortezomib; D045744:Cell Line, Tumor; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007372:Interferons; D007378:Interleukins / Q000378:metabolism*; D053613:Janus Kinase 1 / Q000378:metabolism*; D008545:Melanoma / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D011719:Pyrazines / Q000494:pharmacology; D050791:STAT Transcription Factors / Q000378:metabolism*; D015398:Signal Transduction; D012878:Skin Neoplasms / Q000378:metabolism*; D000077204:Temozolomide; D000095022:Interferon Lambda
D001897:Boronic Acids; D000069286:Bortezomib; D003606:Dacarbazine; D007372:Interferons; D007378:Interleukins; D053613:Janus Kinase 1; D011719:Pyrazines; D050791:STAT Transcription Factors; D000077204:Temozolomide; C000624353:interferon-lambda, human; D000095022:Interferon Lambda