The Role Of MTOR Signal Pathway In Crizotinib-induced Apoptosisi Of EML4-ALK Fusion Gene Positive Lung Adenocarcinoma Cell Line H2228

AIM: mTOR is a member of phosphatidylinositol3-kinase-related kinase(PI3Ks) protein family. Signal pathway with mTOR as the center could play arole in the apoptosis of non-small cell lung cancer (NSCLC) cell lines H2228,which represent positive EML4-ALK fusion gene induced by Crizotinib. So therole of the signal pathway is discussed in this paper.METHODS: H2228cells are processed according to different purposes;Fluorescence quantitative PCR is used to detect gene states; methyl thiazolyltetrazolium assay (MTT assay) is used to detect the cell inhibition rates; flowcytometry (FCM) is used for cell apoptosis and cell cycle; and Western blottesting is adopted to determine the expression and activation levels of the keyproteins in the mTOR signal pathway.RESULTS: Crizotinib has promoted the apoptosis of H2228cells with time anddose dependent. It can block the H2228cells staying at the phase G1. It is foundin apoptotic cell lines processed with Crizotinib that: activation level of mTORis decreased; activation levels of the key proteins in upstream and downstreamof mTOR are both declined; while expression level of fusion protein EML4-ALK V3which has special expression in lung cancer cell lines H2228isnot affected, but its activation form p-ALK is significantly suppressed.CONCLUSION: It is preliminarily confirmed that mTOR signal pathway has acertain relationship with the apoptosis of lung cancer cell H2228whichrepresent positive EML4-ALK fusion gene induced by Crizotinib. And itprovides a basis and reference for the action mechanism of Crizotinib.