| PurposeExtracellular information through the signaling pathway, transmission and regulation of cell life and activities, the study of colon cancer cells in the c-Met signaling pathway pathways and mechanisms, for revealing the nature of tumor invasion and metastasis as well as targeted anti-cancer drugs theoretical basis for clinical application. Use of specific c-Met inhibitor SU11274 as a small molecule indole ring structure with a small molecule compounds of the ATP competitive inhibitor to investigate the cell cycle of colon cancer, and how to reduce the proliferation mechanism of HGF to stimulate induced c-Met phosphorylation and downstream signaling pathway of Ras and PI3 K key molecule Erk, AkT, mTor and the level of rpS6 phosphorylation, thus blocking its mediated effects, which play an inhibitory role of c-Met kinase.MethodThe first chapter 33 cases of colon cancer pathology as the research object to c-Met expression in colon cancer-related, on the c-Met and Its Effects on colon cancer and its downstream signaling pathway expression and immunohistochemical SP Law of 33 cases of colon cancer organizations and the downstream c-Met pathway Erk, Akt rpS6 and mTor protein localization studies. to further explore c-Met signaling and its downstream signal transduction is highly expressed in colon cancer and biological significance.The second and third sections of c-Met selection of specific small molecule inhibitor SU11274 as a research system. the colon cancer cell lines. respectively. k-ras wild-type HT-29. Hct-8 and k-ras mutant Hct-116. DLD-1 as the object of study. using Western blotting. cell proliferation rate, flow three ways to observe the selective C-Met phosphorylation in vitro inhibitor SU11274. HT-29. Hct-8. Hct-116. DLD-1 cells and periodic changes of proliferation, testing the Met and the downstream Erk. Akt rpS6 and mTor signaling pathway related protein phosphorylation. and explore the selective c-Met inhibitor SU11274 against phosphorylation of intracellular signal transduction mechanisms of colon cancer and effect.The fourth section of this combined phosphorylation of c-Met inhibitor SU 11274 and Erk phosphorylation inhibitor PD98059, the colon cancer cell lines in the k-ras gene mutations in Hct-116. DLD-1 as an object of study, using Western blotting, cell proliferation rate, flow three ways to observe the in vitro k-ras gene mutations in Hct-116, DLD-1 cell proliferation and cell cycle, testing the Met and the downstream Erk, Akt rpS6 and mTor associated protein signaling pathway the changes of combination anti-cancer mechanism of intracellular signal transduction and function.ResultsTumor immunohistochemistry results showed that:c-Met.in colorectal cancer, the positive expression rate was 36.3%(12/33); Akt positive expression rate of 51.5% of total c-Met together with the positive expression occurred in 11 cases the positive expression rate was 91.7%:mTOR expression was 60.6% of total c-Met together with the positive expression occurred in 10 cases the positive rate was 83.3%:rpS6 total positive rate was 57.6% and c-Met expression co-occurrence of 11 cases of positive expression was 91.7%; Erk positive rate was 51.5% of total c-Met together with the positive expression occurred in 11 cases the positive rate was 91.7%.SU11274 on the HT-29. HCT-8. Hct-116. DLD-1 cell proliferation results showed that different concentrations of SU11274 were on cultured HT-29, Hct-8. Hct-116. DLD-1 Four cell lines significantly inhibited the proliferation and this effect has dose-dependent and time-dependent, expressed as cell growth slows, reduce the rate of colony forming cells, that in the concentration range. SU11274 the higher the concentration This effect is more obvious.Protein blotting results showed that:colon cancer cell HT-29. Hct-8. Hct-116. DLD-1 under the action of the stimulating factor HGF Met. Akt.. mTor. rpS6. and Erk phosphorylation were significantly increased. Under the action of low concentrations of SU11274 Met phosphorylation has been almost completely suppressed, while diminished Akt, mTor. rpS6. and the level of Erk phosphorylation.Conclusionâ… . there is in colon cancer and c-Met protein and an important protein signaling pathways downstream of Akt. mTor. rpS6. and Erk were abnormally high expression occurs, and the c-Met protein expression and tumor histological type and TNM stage has a certain relevance. â…¡. HGF by binding to its receptor c-Met, the activation of the signaling pathway, the promotion of human colon cancer cell line HT-29, Hct-8, Hct-116, DLD-1 cell growth and proliferation; while enhancing the c-Met pathway, Akt, mTor, rpS6 and protein level of Erk; enhanced cell invasion in vitro.â…¢. c-Met as a target, research and development of small molecule inhibitors SU11274 on human colon cancer cell line HT-29, Hct-8, Hct-116, DLD-1 of the cell cycle, cell proliferation there is a certain role, while the existence of dose and time dependent, and K-ras gene of the wild-type HT-29. Hct-8 cells to suppress better than K-ras gene mutations in Hct-116, DLD-1 cells.â…£. To c-Met as a target of SU11274. and the MEK as a target of combined administration of PD98059, on human colon cancer K-ras gene mutations in Hct-116. DLD-1 cell growth and proliferation were significantly inhibited, and its inhibition of cell proliferation better than single-agent SU11274. and the role of PD98059.
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