The Expression Of Hedgehog Signaling Pathway Gli-1Gene And EMT Related Factor Slug In Cervical Cancer Cells

The integration of Hedgehog signal pathway and abnormal expression of EMT(epithelial mesenchymal transition) related factors with the influence of occurrence of cervical cancer invasion and metastasis, has rarely been reported. Our study investigate the expression inhibits of Hedgehog signaling pathway Gli-1and EMT related factor Slug in cervical cancer Hela cells, by different concentrations of the non-steroidal alkaloids cyclopamine to blocking the Hedgehog signaling pathway on the biological behavior of cervical cancer Hela cells, To explore the possibility of a new target for treatment as cervical cancer of Hedgehog-EMT signal pathway.Objectives:To investigate the expression of Hedgehog signaling pathway Gli-1and EMT related factor Slug in cervical cancer Hela cells, and explore the impact of the Hedgehog signaling pathway and EMT-related factors on the biological behavior of cervical cancer Hela cellsMethods:1. To investigate the cell vitro proliferative capacity and colony formation efficiency of cervical cancer Hela cells by MTT assay and soft agar colony formation assay.2. To detect Gli-1and Slug mRNA expression in cervical cancer Hela cells by RT-PCR.3. To detect Gli-1and Slug protein expression in cervical cancer Hela cells by western blot. Resμlts:1. In MTT assay and soft agar colony formation assay, with increase the drug concentration of Hedgehog signaling pathway inhibitor cyclopamine, the cell proliferation and colony forming ability showed significantly decreased in Hela cells (p<0.05).2. RT-PCR revealed that after treated with different concentration of cyclopamine24h and48h, the expression of Gli-1and Slug genes in the drug group was significantly lower than the control group. The expression of Gli-1and Slug genes was most significant in5uM cyclopamine group. However, in10uM and20uM of cyclopamine group, the expression of Gli-1and Slug genes was gradually decreased. And Gli-1and Slug gene expression dependente on cyclopamine concentration and treated times.3. Western-blot revealed that after treated with different concentration of cyclopamine24h and48h, the expression of Gli-1and Slug proteins in the drug group was significantly lower than the control group,The expression of Gli-1and Slug proteins was most significant in5uM cyclopamine drug group, However, in10uM and20uM of cyclopamine group, the expression of Gli-1and Slug proteins was gradually decreased. And Gli-1and Slug proteins expression dependente on cyclopamine concentration and treated times.4. Conclution:1. Hedgehog signaling pathway inhibitor cyclopamine could effectively inhibit the cell proliferation and colony formation of cervical cancer Hela cell.2. Hedgehog signaling pathway inhibitor cyclopamine could inhibit the gene and protein expression of Hedgehog signaling pathway Gli-1and EMT related factor Slug in cervical cancer Hela cells.