Effect Of Polydatin On Epithelial-Mesenchymal Transition Of A549Cells Induced By TGF-β1

Objective To study the effects of Polydatin on the growth of A549cells and expression of E-cad、Vimintin in A549cells induced by TGF-β1. Explore the mechanism of Polydatin restraining the process of EMT induced by TGF-β1. Provides the basis for the clinical medication.Method The A549cells were cultured in serum-free medium were randomly divided into five groups after24h.Take a group as blank group to join the culture medium, the other one as control group containing only added TGF-β1(final concentration5ng/ml) in the culture medium.The remaining three groups as low-dose group, middle dose group,high dose group and were added to the same volume containing polydatin of different concentrations (50,100,150umol/ml) and TGF-β1(final concentration5ng/ml) culture medium.①Observing cellular morphology changes by inverted phase-contrast microscope and recording the changes after A549cells being treated24h and48h.②Through MTT method to detect different concentrations of polydatin effect on A549cells EMT process induced by TGF-β1.Then get the optimal concentration.③Western-blot assay the protein expression of epithelial cell marker E-cad, mesenchymal cell marker Vimentin of blank group, control group, low dose group, middle dose group at24h,48h. RT-PCR assay the mRNA expression of epithelial cell marker E-cad, mesenchymal cell marker Vimentin of blank group, control group, low dose group, middle dose group at24h,48h.Result Induced by TGF-β1, cellular morphology of A549cells turn from pebble shape to fusiform shape.The connection between cells becomes loose. After adding polydatin the inhibition rate of A549cells induced by TGF-β1:24h:low-dose group, middle dose group, high dose group were:37.72%,46.44%,47.69%;48h:low dose group, middle dose group, high dose group were:38.73%,49.35%,49.52%. Protein and mRNA expression of epithelial marker E-cad showed a descending tendency (P<0.05).But compared with the control group, each drug group showed a lesser descending tendency (P<0.05). Mesenchymal marker Vimentin showed an ascending tendency (P<0.05). But compared with the control group, each drug group showed a lesser ascending tendency (P<0.05). Contrast with24h,48h is obvious.Conclusion①Polydatin could maintain epithelial morphology of A549cells induced by TGF-β1, inhibiting turning to mesenchymal morphology;(2)The highest inhibition rate is middle dose group③At the protein and mRNA levels, descending tendency of epithelial marker E-cad and ascending tendency of mesenchymal marker Vimentin were48h and middle dose group obvious, reflecting the time-dependence and dose-dependence of polydatin in inhibition of EMT.