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Inhibitors of Ras/Raf-1 interaction identified by two-hybrid screening revert Ras-dependent transformation phenotypes in human cancer cells

Authors
  • Juran Kato-Stankiewicz
  • Irina Hakimi
  • Gang Zhi
  • Jie Zhang
  • Ilya Serebriiskii
  • Lea Guo
  • Hironori Edamatsu
  • Hiroshi Koide
  • Sanjay Menon
  • Robert Eckl
  • Sukumar Sakamuri
  • Yingchun Lu
  • Quin-Zene Chen
  • Seema Agarwal
  • William R. Baumbach
  • Erica A. Golemis
  • Fuyuhiko Tamanoi
  • Vladimir Khazak
Publisher
National Academy of Sciences
Publication Date
Oct 21, 2002
Source
PMC
Keywords
Disciplines
  • Biology
  • Medicine
License
Unknown

Abstract

The interaction of activated Ras with Raf initiates signaling cascades that contribute to a significant percentage of human tumors, suggesting that agents that specifically disrupt this interaction might have desirable chemotherapeutic properties. We used a subtractive forward two-hybrid approach to identify small molecule compounds that block the interaction of Ras with Raf. These compounds (MCP1 and its derivatives, 53 and 110) reduced serum-induced transcriptional activation of serum response element as well as Ras-induced transcription by way of the AP-1 site. They also inhibited Ras-induced Raf-1 activation in human embryonic kidney 293 cells, Raf-1 and mitogen-activated protein kinase kinase 1 activities in HT1080 fibrosarcoma cells, and epidermal growth factor-induced Raf-1 activation in A549 lung carcinoma cells. The MCP compounds caused reversion of ras-transformed phenotypes including morphology, in vitro invasiveness, and anchorage-independent growth of HT1080 cells. Decreased level of matrix metalloproteinases was also observed. Further characterization showed that MCP compounds restore actin stress fibers and cause flat reversion in NIH 3T3 cells transformed with H-Ras (V12) but not in NIH 3T3 cells transformed with constitutively active Raf-1 (RafΔN). Finally, we show that MCP compounds inhibit anchorage-independent growth of A549 and PANC-1 cells harboring K-ras mutation. Furthermore, MCP110 caused G1 enrichment of A549 cells with the decrease of cyclin D level. These results highlight potent and specific effects of MCP compounds on cancer cells with intrinsic Ras activation.

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