The Study Of The Epithelial-mesenchymal Transition Induced By TGF-β1 Mediated PI3K/Akt Signaling Pathway In Cervical Cancer

Objective: To figure out whether the Epithelial-mesenchymal Transition(EMT)induced by TGF-β1-mediated PI3 K / Akt Signaling Pathway and enhanced the ability of migration and invasiveness of Cervical Cancer.Method: The cultured cells were divided into four groups in the experiment.(1)control group: adding the same amount of PBS as control.(2)TGF-β1 Group: adding TGF-β1 with a concentration of 10 ng / ml in Hela cells.(3)TGF-β1 + LY294002 group: adding 10μmol / L LY294002 and then add TGF-β1 in Hela cells.(4)LY294002 group: adding 10μmol / L LY294002 in Hela cells.1.Fouty-eight hours after treatment,mrophological change of human cervical carcinoma cells in each group were observed by inverted microscope.2.The migration and invasiveness ability of three group(control group、TGF-β1 Group and TGF-β1 + LY294002 group)was detected by transwell experiment.3.Western blot was used to detect the expression of E-cadherin、vimentin、p-PI3 K and p-Akt(Thr308)in the the four group cells.Results: 1.Cells in the control group exhibit typical epithelial cell features.Cells in the TGF-β1 group was charcterized by fusiformis appearence.Cells in the LY294002 group and TGF-β1+LY294002 group were gathered into a group,circular or elongated shuttle type,do not exhibit significant mesenchymal cell-like phenotype.2.migration experiment found that the migration number of cells in TGF-β1 group(67.0 ± 7.0)cells increased significantly than the control group(39.5 ± 4.4)cells,the migration number of cells in LY294002 + TGF-β1 were(51.3 ± 2.2)cells decreased significantly than that of TGF-β1 group,there were significantly difference between them(P <0.01).Invasion assay results showed that the invasion number of cells in TGF-β1 group were increased than that of the control group(31.5 ± 2.1)cells and LY294002 + TGF-β1 group(42.8 ± 3.3)cells,the difference was statistically significant(P <0.01).3.Western blot showed that(1)the expression of p-PI3 K and p-Akt in untreated human cervical carcinoma Heala cells were low,and after treatment with TGF-β1,the expression of p-PI3 K and p-Akt were significantly increased and after treated with LY294002 and LY294002 + TGF-β1 the expression of p-PI3 K and p-Akt were significantly decreased(P<0.05).(2)In untreated group the expression of E-cadherin was high while vimentin was low in Hela cells,E-cadherin was significantly decreased and vimentin was remarkly increased after treatment by TGF-β1.However,After treated by LY294002 and LY294002 + TGF-β1,which significantly up-regulation the expression of E-cadherin and down-egulation the expression of vimentin(P<0.05).Conclusion: 1.TGF-β1 can help Hela cells to obtain some characteristics of stromal cell and promote EMT.2.After treated by TGF-β1 in Hela cells,the PI3 K / Akt signaling pathway were activated and EMT were promoted,the migration and invasion ability of Hela cells were increased.