The Role Of TGF-β1in Regulation Of Pokemon Expression In Breast Cancer Cell

Objective: To reveal the effect of TGF-β1on the expression of Pokemon in breastcancer cells, and to clarify the potential underlying mechanisms.Methods: MCF-7and MDA-MB-231cells were used in this work. Experiment wasdivided into three parts.1) MCF-7and MDA-MB-231cells were treated withdifferent concentrations of TGF-β1for a certain time or the same concentrationTGF-β1for different time, then RT-PCR technology, immunofluorescence stainingtechnique and Western blot analysis were used to detect the expression of Pokemon.2)To verify the role of PI3K/Akt signaling pathway in TGF-β1induced Pokemonexpression. MCF-7and MDA-MB-231cells were pretreated with PI3K/Akt signalingpathway inhibitor LY294002for2h，then treated with TGF-β1for9h,RT-PCRtechnology and immunofluorescence staining were used to detect the role ofPI3K/Akt signaling pathway in TGF-β1induced Pokemon expression.3) The effect ofTGF-β1on Pokemon promoter activity in MCF-7cells. The PGL4.10Pokemonpromoter plasmid transfected MCF-7cells were treated with different concentrationsof TGF-β1for24h, and luciferase reporter assay were used.Results:1) The effect of TGF-β1in breast cancer cells on the expression ofPokemon.①Compared with the control group, TGF-β1increase the level ofPokemon mRNA in MCF-7cells, and reach the peak at the concentration of5ng/mlTGF-β1group(P<0.01); Compared with the control group, TGF-β1increase the levelof Pokemon mRNA in MCF-7cells, and reach the peak at the time point of12h(P<0.01).②Compared with the control group, TGF-β1increase the level ofPokemon protein in MCF-7cells, and reach the peak at the concentration of5ng/mlTGF-β1group(P<0.05);2) The role of PI3K/Akt signaling pathway in TGF-β1induced the expression of Pokemon in breast cancer cells.①LY294002(10μM and20μM) could inhibit the expression of Pokemon mRNA transcription level induced by TGF-β1in MCF-7and MDA-MB-231cells (P<0.05).②LY294002(10μM) couldblock TGF-β1induced Pokemon the mean fluorescence intensity of Pokemon was inboth MCF-7and MDA-MB-231cells (P<0.05).③The result of western blot:compared with the control group, TGF-β1could increase Pokemon level and reachedthe peak at time of6h,olso could increase P-Akt level and reached the peak at time of30min in MCF-7and at time of1h in MDA-MB-231cells(P<0.01).3) The result ofpromoter luciferase activity: compared with the control group, TGF-β1couldincrerease the promoter activity of Pokemon in MCF-7cells (P<0.05).Conclusion:1) TGF-β1could promote the expression of Pokemon in breastcancer cells.2) PI3K/Akt signaling pathway might be involved in TGF-β1inducedPokemon expression in breast cancer cells.