Premium
Genomewide RNA i screen identifies protein kinase C β and new members of mitogen‐activated protein kinase pathway as regulators of melanoma cell growth and metastasis
Author(s) -
Schönherr Madeleine,
Bhattacharya Animesh,
Kottek Tina,
Szymczak Silke,
Köberle Margarethe,
Wickenhauser Claudia,
Siebolts Udo,
Saalbach Anja,
Koczan Dirk,
Magin Thomas M.,
Simon Jan C.,
Kunz Manfred
Publication year - 2014
Publication title -
pigment cell and melanoma research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.618
H-Index - 105
eISSN - 1755-148X
pISSN - 1755-1471
DOI - 10.1111/pcmr.12216
Subject(s) - melanoma , mapk/erk pathway , biology , cancer research , gene knockdown , protein kinase a , protein kinase c , microbiology and biotechnology , cell growth , mitogen activated protein kinase , rna , kinase , cell culture , gene , genetics
Summary A large‐scale RNA i screen was performed for eight different melanoma cell lines using a pooled whole‐genome lentiviral sh RNA library. sh RNA s affecting proliferation of transduced melanoma cells were negatively selected during 10 days of culture. Overall, 617 sh RNA s were identified by microarray hybridization. Pathway analyses identified mitogen‐activated protein kinase ( MAPK ) pathway members such as ERK 1/2, JNK 1/2 and MAP 3 K 7 and protein kinase C β ( PKC β) as candidate genes. Knockdown of PKC β most consistently reduced cellular proliferation, colony formation and migratory capacity of melanoma cells and was selected for further validation. PKC β showed enhanced expression in human primary melanomas and distant metastases as compared with benign melanocytic nevi. Moreover, treatment of melanoma cells with PKC β‐specific inhibitor enzastaurin reduced melanoma cell growth but had only small effects on benign fibroblasts. Finally, PKC β‐sh RNA significantly reduced lung colonization capacity of stably transduced melanoma cells in mice. Taken together, this study identified new candidate genes for melanoma cell growth and proliferation. PKC β seems to play an important role in these processes and might serve as a new target for the treatment of metastatic melanoma.