| Objective: To investigate whether transforming growth factor-β1(TGF-β1)regulates the proliferation,invasion,migration and epithelialization of chorionic cancer cell line JEG-3 through PI3K/AKT signaling pathway.Methods: Chorionic cancer cell line JEG-3 was cultured in vitro and randomly divided into control group and TGF-β1 group(TGF-β1 concentration: 25,50,100 and200ng/m L),after treated for 0h,24 h,48h and 72 h,the proliferation capacity of JEG-3cells was determined by CCK-8 cell proliferation assay,which was used to screen and determine the optimal concentration and time of TGF-β1.The cell scratch assay and Transwell cell migration assay were used to detect the effect of the control group and the100ng/ml TGF-β1 group on the migration ability of JEG-3 cells after 48 h treatment.Transwell invasion assay was used to detect the effects of the control group and the100ng/ml TGF-β1 group on JEG-3 cell invasion.The expression of PI3 K,p-PI3 K,AKT,p-AKT,E-cadherin and Vimentin in JEG-3 cells induced by 100ng/ml TGF-β1 was detected by Western blot.Functional recovery experiment: JEG-3 cells were cultured and divided into control group,TGF-β1 group and LY294002+TGF-β1 group(JEG-3 cells were pretreated with 20μmol/L LY29400 for 1h,and then 100ng/ml TGF-β1 was added for further culture).The above experiments were repeated to verify the effects of TGF-β1on proliferation,migration and invasion of choriocarcinoma and the changes in protein expression.Results: 1.The results of CCK-8 experiment showed that the proliferative activity of JEG-3 cells was gradually enhanced with the increase of TGF-β1 concentration and time.Compared with the concentration of 100ng/ml,the concentration of 200ng/ml did not increase significantly.The results showed that TGF-β1 had a good concentration and time dependence on the proliferation ability of chorionic cancer cells within a certain concentration range,and the difference was statistically significant(P<0.05).Since the increasing trend of concentration was more obvious at 100ng/ml and 48 h,it was used as the concentration and time for subsequent experiments.2.Scratch test showed that the mobility of JEG-3 cells in the control group and TGF-β1 group was(46.31±0.024)% and(80.92±0.038)%,respectively,after 48 h treatment,and the difference was statistically significant(P<0.01).3.The results of Transwell cell migration experiment showed that the number of cells passing through the cells with(765.5±43.13)microscopes in TGF-β1 group was significantly increased compared with the control group(383.5±12.02)microscopes per high-power microscope.The difference was statistically significant(P<0.05).4.The results of Transwell chamber invasion experiment showed that the number of cells passing through the chamber was significantly increased in TGF-β1 group(883.5±45.96)compared with the control group(342.0±16.97)cells/high power microscope.The difference was statistically significant(P<0.01).5.Western blot results showed that the expression levels of PI3 K and AKT in100ng/ml TGF-β1 treatment group were not significantly changed compared with the control group,with no statistical significance(P>0.05).The expression levels of p-PI3 K,p-AKT,E-cadherin and Vimentin in 100ng/ml TGF-β1 treatment group were(1.986±0.0014,2.2405±0.0063,0.6995±0.0615,1.2055±0.0389),respectively.Compared with the control group(0.9985±0.0035,0.9985±0.0063,0.9765±0.0063,1.0285±0.0078),the differences were statistically significant(P<0.05).6.Functional recovery experiment: After adding PI3 K inhibitor(20umol/L LY294002),TGF-β1 inhibited the proliferation,migration,invasion and epithelial interstitial transformation(EMT)of JEG-3 cells;Western blotting showed that the addition of LY294002 inhibited the phosphorylation of TGF-β1 to PI3 K and AKT.Conclusion:1.Exogenous TGF-β1 showed concentration and time dependence on the proliferation of chorionic cancer cell line JEG-3.2.TGF-β1 can promote the proliferation,migration,invasion and EMT of chorionic cancer cells.3.TGF-β1 may promote the malignant biological behavior of chorionic cancer cells through PI3K/AKT signaling pathway. |
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