Mechanism Analysis Of EZH2 Regulating Hedgehog-Gli Signaling Pathway In Malignant Pleural Mesothelioma

Research Background:Malignant pleural mesothelioma(MPM),as a more common tumor in the world,has high invasiveness and poor prognosis,and has limited clinical treatment.The EZH2 gene is a chromatin-modified transcriptional repressor that has been associated with the development and prognosis of various malignant tumors such as lung cancer in recent years.The functional role and pathway mechanism of EZH2 in MPM have also gradually developed.At present,the pathogenesis of MPM has not been fully elucidated.Studies have shown that the activation of Hedgehog-Gli signaling pathway plays an important role in the development of MPM.Related experiments show that in MPM,as E3 ubiquitin ligase's core member Cul4 A,whose initiation sequence is bound by EZH2,and Cul4 A activates the Hedgehog-Gli1 pathway via mTOR,therefore we speculate that there is a correlation between EZH2 and Hedgehog-Gli1 signaling pathway in MPM,ie EZH2 is dependent on mTOR signal in MPM Thereby regulating the Hedgehog-Gli1 signaling pathway.This study intends to investigate whether EZH2 regulates Hedgehog-Gli signaling pathway in MPM depends on mTOR signal,and elucidates the mechanism of action of EZH2 in malignant pleural mesothelioma.Objective:To explore the molecular mechanism of EZH2 in regulating Hedgehog signaling pathway in malignant pleural mesothelioma,to elucidate the mechanism of EZH2 in the development of malignant pleural mesothelioma,and to find the internal pathway and potential molecular targets for the treatment of malignant pleural mesothelioma.Methods:1.Conventional culture of EZH2 and Gli1 low expression cell lines(MSTO-211H).2.The amplification and extraction of the recombinant plasmid and the empty plasmid,and the EZH2 gene were introduced to obtain a cell line with high expression of EZH2.3.The expression of Bcl-1 gene was increased in MTO cell line MSTO-211 H,which was highly expressed in EZH2.The expression of Bcl-1 was decreased after the addition of mPAR signaling inhibitor Rapamycin.4.The mRNA and protein expression of Gli1?Gli1 upstream genes N-Myc and Gli1 downstream regulatory genes,P21 and Bcl-1 genes were detected by Real-time PCR and Western blot.Results:1.The introduction of EZH2 gene into MPM cell line MSTO-211 H can increase the expression of Gli1.After the addition of mTOR signal inhibitor Rapamycin,the expression of Gli1 did not change significantly.2.The EZH2 gene was introduced into MPM cell line MSTO-211 H,which could make P21 low expression.After the addition of mTOR signal inhibitor Rapamycin,the expression of P21 increased.3.The introduction of EZH2 gene into MPM cell line MSTO-211 H can increase the expression of Bcl-1.After the addition of mTOR signal inhibitor Rapamycin,the expression of Bcl-1 is decreased.4.The expression of N-Myc was not significantly changed in the MPM cell line MSTO-211 H.The expression of N-Myc gene was decreased after the addition of the mTOR signal inhibitor Rapamycin.Conclusion:EZH2 may activate the Hedgehog-Gli signaling pathway in malignant pleural mesothelioma,but its activation mechanism may be independent of mTOR signaling.