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

154 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

Fingerprint

Lentivirus
RNA Interference
Melanoma
Growth
Mitogen-Activated Protein Kinases
Mutation
Cell Line
Neoplasms
Matrix Metalloproteinase 1
Integrins
Genetic Therapy
Small Interfering RNA
Signal Transduction
Phosphotransferases
Phosphorylation
HIV
Phenotype

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research
  • Genetics

Cite this

Sumimoto, H., Miyagishi, M., Miyoshi, H., Yamagata, S., Shimizu, A., Taira, K., & Kawakami, Y. (2004). Inhibition of growth and invasive ability of melanoma by inactivation of mutated BRAF with lentivirus-mediated RNA interference. Oncogene, 23(36), 6031-6039. https://doi.org/10.1038/sj.onc.1207812

Inhibition of growth and invasive ability of melanoma by inactivation of mutated BRAF with lentivirus-mediated RNA interference. / Sumimoto, Hidetoshi; Miyagishi, Makoto; Miyoshi, Hiroyuki; Yamagata, Shizuko; Shimizu, Ayako; Taira, Kazunari; Kawakami, Yutaka.

In: Oncogene, Vol. 23, No. 36, 12.08.2004, p. 6031-6039.

Research output: Contribution to journalArticle

Sumimoto, H, Miyagishi, M, Miyoshi, H, Yamagata, S, Shimizu, A, Taira, K & Kawakami, Y 2004, 'Inhibition of growth and invasive ability of melanoma by inactivation of mutated BRAF with lentivirus-mediated RNA interference', Oncogene, vol. 23, no. 36, pp. 6031-6039. https://doi.org/10.1038/sj.onc.1207812
Sumimoto, Hidetoshi ; Miyagishi, Makoto ; Miyoshi, Hiroyuki ; Yamagata, Shizuko ; Shimizu, Ayako ; Taira, Kazunari ; Kawakami, Yutaka. / Inhibition of growth and invasive ability of melanoma by inactivation of mutated BRAF with lentivirus-mediated RNA interference. In: Oncogene. 2004 ; Vol. 23, No. 36. pp. 6031-6039.
@article{205bb6a7db4442d9aad6ae073bd92108,
title = "Inhibition of growth and invasive ability of melanoma by inactivation of mutated BRAF with lentivirus-mediated RNA interference",
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.",
keywords = "BRAF, Gene therapy, Lentiviral vector, MAPK, Melanoma, RNAi",
author = "Hidetoshi Sumimoto and Makoto Miyagishi and Hiroyuki Miyoshi and Shizuko Yamagata and Ayako Shimizu and Kazunari Taira and Yutaka Kawakami",
year = "2004",
month = "8",
day = "12",
doi = "10.1038/sj.onc.1207812",
language = "English",
volume = "23",
pages = "6031--6039",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "36",

}

TY - JOUR

T1 - Inhibition of growth and invasive ability of melanoma by inactivation of mutated BRAF with lentivirus-mediated RNA interference

AU - Sumimoto, Hidetoshi

AU - Miyagishi, Makoto

AU - Miyoshi, Hiroyuki

AU - Yamagata, Shizuko

AU - Shimizu, Ayako

AU - Taira, Kazunari

AU - Kawakami, Yutaka

PY - 2004/8/12

Y1 - 2004/8/12

N2 - 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.

AB - 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.

KW - BRAF

KW - Gene therapy

KW - Lentiviral vector

KW - MAPK

KW - Melanoma

KW - RNAi

UR - http://www.scopus.com/inward/record.url?scp=4444240325&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=4444240325&partnerID=8YFLogxK

U2 - 10.1038/sj.onc.1207812

DO - 10.1038/sj.onc.1207812

M3 - Article

VL - 23

SP - 6031

EP - 6039

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 36

ER -