The Effect And Mechanisms Of MiR-101 Inhibits The Proliferation And Metastasis Of Lung Cancer By Targeting Zinc Finger E-box Binding Homeobox 1

Background:Non-small cell lung cancer(NSCLC)is the main pathological type of lung cancer,accounting for 85% of the total number of patients with lung cancer,and the 5-year survival rate is low.In recent years,with the rapid development of molecular biology,the early detection rate and therapeutic effect of NSCLC patients have been greatly improved.However,some of the molecular mechanisms that induce disease still lack deep understanding,and some patients are not satisfied with the treatment results.Many miRNAs are involved in the occurrence and development of lung cancer,and miR-101,through negative regulation of cell proliferation and metastasis,shows that it may be a key regulatory molecule in the development of NSCLC.At present,the potential molecular mechanism of miR-101 in NSCLC is largely unknown.Purpose:To explore the expression differences and prognostic correlation of miR-101 andE-box binding homeobox 1(ZEB1)between NSCLC tissues and adjacent tissues;Through vivo and vitro experiments,the expression of miR-101 and ZEB1 were regulated,and to explore the mechanism of miR-101 targeting ZEB1 inhibiting the growth and metastasis of NSCLC.Methods:1.The expression of miR-101 and ZEB1 mRNA in NSCLC tissues and adjacent tissues was detected by RT-PCR,and the expression of ZEB1 protein in NSCLC tissues and adjacent tissues was detected by Western Blot.2.The expression of miR-101 in NSCLC cell lines and normal human bronchial epithelial cell line were detected by RT-PCR,and the expression of ZEB1 protein in NSCLC cell lines and normal human bronchial epithelial cell line were detected by Western Blot.3.By up-regulating the expression of miR-101 and ZEB1 in A549 cell line,the differences in cell growth and migration ability were observed in vitro.4.By up-regulating miR-101 and down-regulation of ZEB1 expression,the differences in subcutaneous tumor growth and lung tumor metastasis of SCID mice were observed in vivo.5.The complementary sequences of miR-101 and ZEB1 were detected by TargetScan database.ZEB1 was confirmed to be the target gene of miR-101 by the Dual-luciferase reporter assay.6.Through Western Blot,the mechanism of miR-101 targeted regulation of ZEB1 inhibiting the growth and metastasis of NSCLC was further explored.Results:1.The expression of miR-101 in NSCLC tissues and cell lines was significantly lower than that of the adjacent tissues and normal bronchial epithelial cells,and the expression of ZEB1 was the opposite.The expression of miR-101 was negativelycorrelated with lymph node metastasis.2.The up-regulated expression of miR-101 alone could inhibit proliferation,apoptosis resistance,migration and invasion of NSCLC cells,while up-regulating both miR-101 and ZEB1 expression were significantly reduced.3.The up-regulated expression of miR-101 inhibits tumor growth and metastasis in SCID mice,and the down-regulation expression of ZEB1 can also produce a similar inhibitory effect.4.The Dual-luciferase reporter assay suggested that ZEB1 was a direct target of miR-101,and miR-101 inhibited the malignant phenotype of NSCLC by targeting ZEB1.The main mechanism is: miR-101 inhibits the occurrence of ZEB1 mediated EMT;By down-regulated the expression of CyclinD1,up-regulated the expression of Rb,cell roliferation was inhibited;By down-regulated the expression of bcl-2,up-regulated the expression of Bax,increased the activity of Caspase-3,cell apoptosis was promoted.Conclusions:This study shows that miR-101 inhibits the growth and metastasis of NSCLC by targeting ZEB1,suggesting that miR-101 is a key molecule in NSCLC growth and metastasis,and can provide a new way for the treatment of NSCLC.