The Interaction Mechanism Between LSD1 And MTOR Pathway In Esophageal Squamous Cell Carcinoma

Experimental Purposes1. To study the molecular regulatory mechanism between LSD1 and mTOR in esophageal squamous cell carcinoma cell lines.2. To study the effects of LSD1 inhibitor on esophageal squamous cell carcinoma cell proliferation and its mechanism so as to provide a theoretical basis for targeted therapy of esophageal squamous cell carcinoma.Experimental Methods 1. The protein expression associated with LSD1 and m TOR pathway in different esophageal squamous cell carcinoma cell lines.Western blot assay detects the expression levels of above pathway protein LSD1, PTEN, p-AKT(ser473), p-p70S6 K, Rictor, Raptor in different esophageal squamous cell carcinoma cell lines.2. Effects of LSD1 inhibitor on TE-1, ECa109 cell proliferation.CCK-8 assay estimates the proliferation inhibition effects of LSD1 inhibitor in esophageal squamous cell carcinoma cell line TE-1, ECa109.3. The effects of inhibiting LSD1 to mTOR and histone.Western blot assay detects the expression levels of LSD1, PTEN, p-p70S6 K, Rictor, Raptor proteins and histone H3K4me2, H3K9me2 after treatment with LSD1 inhibitor and sh RNA-LSD1 in ESCC cell lines ECa109 for 24 h.4. The effects of inhibiting mTOR pathway to LSD1.Western blot assay detects the protein expressions of LSD1, Rictor, Raptor, p-p70S6 K after esophageal squamous cell carcinoma cells treated with different concentrations of ATP competitive m TORC1/m TORC2 dual inhibitors PP242(0,1,5,10μM)and m TORC1 inhibitor rapamycin rapamycin(0,50,100,200 n M) for 24 h.Experimental Results 1. The expression of LSD1 can be detected in the five selected esophageal squamous cell carcinoma cells in which ECa109 have higher LSD1 expression.2. LSD1 inhibitors significantly inhibited cell proliferation of TE-1, ECa109. m TORC1, m TORC2 are subject to different degree’s inhibition after treatment with cytostatic for 24h; LSD1 expression did not change, but its substrate H3K9 dimethylation levels elevated. sh RNA-LSD1 interference using to transfect ECa109 cells resulted in reduced expression of protein associated with m TOR.3. ECa109 cells treated with rapamycin or pp242 were detected the reduced expression of LSD1.Conclusions 1. LSD1 has different expression levels in five different esophageal squamous cell carcinoma TE-1、ECa109、KYSE450、KYSE750、EC9706.2. LSD1 inhibitors can inhibit cell proliferation.3. inhibition of LSD1 can inhibit the expression of mTOR pathway proteins which proved that LSD1 is capable of modulating m TOR pathway.Inhibition of m TOR pathway can lead to downregulation of LSD1 expression, proving that m TOR can regulate LSD1.