Screen Metastasis-associated Genes From Anoikis Resistant A549 Lung Cancer Cells By Human Genome Array And Effect Of TC-1 Singal Passway Inhibitor On Anoikis

In 1994,Frisch SM reported a special form of programmed cell death(apoptosis) called"anoikis"[1],which induced by the deteach of epithelial cell from their normal Extracellular matrix (ECM). The biological propose of anoikis is that it provent deteached epithelial cells from keep growing in abnormal place through vascular system .However,most tumor cells lose this feature, exfoliated from Primary site, migrate into distant organ and keep growing with abnormal ECM[2].This phenomenon of"anoikis resistant"in some tumor is one of the important reason that metastatic tumor occur.The phenomenon of"anoikis/ anoikis resistant"involves with complex Signal transduction. The essence of anoikis resistant in some tumor cells is abnormality of anoikis-mediated signal pathways. Researches have been done on the differences in signal transduction processes between adherent and suspension cultured tumor cells.It is clear that integrin-associated signal pathways, death receptor signal pathways, and the cytoskeleton,overexpression of Oncogene ras, raf, rac, src , and inhibition of suppressor gene PTEN, p53 play important roles in anoikis resistant[3-4]. however, the systemic mechanism of anoikis / anti-anoikis is still unclear systematically, The diversity of cells and mutation-prone nature of tumor cells made the different tumors may have different mechanism of anoikis resistant.Objective This experiment tries to screen metastasis-associated genes from anoikis resistant and adherent growth A549 lung cancer cell by Human Genome Array, inspect and verify the effect on the anoikis by TC-1-related pathway inhibitor.Methods Establish anoikis resistant A549 lung cancer cell lines by using poly-hydroxyethyl methacrylate resin processed petri dishes,which courses cell free from adherent. Testing the different expressed gene between anoikis resistant A549 cell and adherent growth A549 cell,using human V2.0 whole-genome oligonucleotide microarray,a product of Capitalbio Corporation,Beijing. Screen metastasis-associated genes. Both of the cells are treated with TC-1-related pathway inhibitor PD176074,a Sigma-Aldrich product, verify the effect of the inhibitor on anoikisResults 745 different expressed genes were screened,including 38 highly metastasis-associated genes.PD176074 can decreased the ability of anoikis resistant in A549 cells.Conclusion The successfully established anoikis resistant A549 cell lines and screened different expressed genes , and it approved that TC-1-related pathway inhibitor PD176074 can decreased the ability of anoikis resistant in A549 cells.This provides us basis for futher research on metastasis of lung cancer.