The Role Of Pokemon In Proliferation And Migration Of MDA-MB-231 Cells

Objective:To explore the effect of Pokemon on the proliferation and migration of MDA-MB-231 cells,and to unravel the related molecular mechanisms.Methods:The MDA-MB-231 cell maintains high expressing Pokemon was adopted as the research object.Using MTT experiment method to detect the proliferation of MDA-MB-231 cells and to draw the growth curve.At the same time,via plate clone formation experiment to prove the effect of Pokemon on the proliferation of breast cancer cells.Then,the relationship between overexpression of Pokemon and MDA-MB-231 cell cycle distribution was detected by the flow cytometry.The breast cancer cell MDA-MB-231 in vitro culture was selected as cell models,Luciferasy assay was used to detect the impact of Pokemon on the promoter activity of CyclinA.Through Western Blot experiment method to learn the influence of Pokemon on expression of CylinA from the perspective of protein level.Transwell test was utilized to determine the impact of pokemon on the invasion and metastasis of MDA-MB-231 cells.The influence of Pokemon on the protein expression level of the epithelial cell markers(Clandin-1,ZO-1)and mesothelial molecules(Vimentin and TCF-8)was tested by the Western Blot.The breast cancer cell MDA-MB-231 in vitro culture was selected as cell models,using flow cytometry to select two groups of cells,including CD44high/CD24 low and CD44low/CD24 high,from the MDA-MB-231 cells.qRT-PCR technology was used to compare Pokemon,TGF-β1 and Smand4 m RNA transcriptional level in the two groups of cells.Results:1)MTT experiment suggested Pokemon could inhibit the proliferation of MDA-MB-231 cells.2)Pokemon could impair the MDA-MB-231 cell colony formation ability.3)Pokemon could influence the periodical distribution of MDA-MB-231 cells,reduce number of cells in S phase and increase in G2.4)Pokemon inhibited the promoter activity of CyclinA in MDA-MB-231 cells.5)Pokemon decreased the expression of CyclinA protein.6)Transwell experimental results showed that Pokemon could restrain the migration ability of MDA-MB-231 cells.7)Pokemon promoted the expression of the epithelial cell markers(Clandin-1,ZO-1),but inhibited mesothelial molecules(Vimentin and TCF-8).8)The mRNA transcription level of Pokemon in the CD44low/CD24 high cell group was more significant than that of the CD44high/CD24 low cell group.However,the mRNA transcription level of TGF-β1 and Smand4 is significantly lower than that of the CD44high/CD24 low group.Conclusion:1)Pokemon inhibited proliferation of MDA-MB-231 cells,.reduce number of cells in S phase and increase in G2.Pokemon might be inhibit the expression of CyclinA by suppressing the promoter activity of it in MDA-MB-231 cells.2)Pokemon could inhibit the migration ability of MDA-MB-231 cells by inhibiting the transformation of epithelioid cells to mesenchymal cells.