Inhibition of growth and invasive ability of melanoma by inactivation of mutated BRAF with lentivirus-mediated RNA interference

Hidetoshi Sumimoto, Makoto Miyagishi, Hiroyuki Miyoshi, Shizuko Yamagata, Ayako Shimizu, Kazunari Taira, Yutaka Kawakami

Research output: Contribution to journalArticle

158 Citations (Scopus)

Abstract

Oncogenic mutations of molecules involved in the mitogen-activated protein kinase (MAPK) pathways provide signals mediating both tumor growth and invasion in various cancers including melanomas. BRAF somatic mutations, found in 66% of melanomas, have NIH3T3 transforming ability with the elevated kinase activity in vitro. We attempted to mediate RNA interference (RNAi) with HIV lentiviral vectors specific for either wild type or the most frequently mutated form of BMAF (V599E) in 10 melanoma cell lines, and found that RNAi inhibited the growth of most melanoma cell lines in vitro as well as in vivo, which was accompanied by decrease of both BRAF protein and ERK phosphorylation. Interestingly, the mutated BRAF (V599E)-specific siRNA inhibited the growth and MAPK activity of only melanoma cell lines with this mutation. Furthermore, BRAF RNAi inhibited matrigel invasion of melanoma cells accompanied with a decrease of matrix metalloproteinase activity and β1 integrin expression. These results clarify that the mutated BRAF (V599E) is essentially involved in malignant phenotype of melanoma cells through the MAPK activation and is an attractive molecular target for melanoma treatment. The lentivirus-mediated RNAi specific for oncogenic mutations may be a powerful technique for gene therapy of cancer.

Original languageEnglish
Pages (from-to)6031-6039
Number of pages9
JournalOncogene
Volume23
Issue number36
DOIs
Publication statusPublished - 2004 Aug 12

Keywords

  • BRAF
  • Gene therapy
  • Lentiviral vector
  • MAPK
  • Melanoma
  • RNAi

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

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