Effect Of Retinol Binding Protein 4 On Migration And Invasion Of Non-Small Cell Lung Cancer

Lung Cancer is one of the most common tumors in the world,accounting for 11.6% of all cases.Non-small cell lung cancer(NSCLC)is the main form of lung cancer,accounting for 85% of lung cancer,which is characterized by easy metastasis,easy recurrence,poor prognosis and so on.In recent years,with the emergence of molecular targeted therapy and immunotherapy,the treatment of NSCLC has made great progress.Unfortunately,drug resistance is inevitable in many patients,so it is important to find new therapeutic targets.Retinol Binding Protein4(RBP4)is a new adipokine that has been widely studied in recent years and belongs to the Retinol family.RBP4 has a variety of biological functions,and the relationship between RBP4 and type 2 diabetes and cardiovascular disease was mainly studied at the earliest.However,in recent years,researchers have begun to study the relationship between the disease and cancer.Several studies have shown that RBP4 plays a specific role in liver,ovarian,breast,and colorectal cancer,but there has been little research on RBP4 and non-small cell lung cancer.This study focused on the effect of RBP4 on the proliferation,migration and invasion function of NSCLC,aiming to explore the mechanism of RBP4 on NSCLC.Purpose: In this study,human normal lung epithelial cell BEAS-2B and human non-small cell lung cancer cell A549 and nude mouse tumor-forming model were used as research objects.Firstly,the difference in RBP4 expression levels between lung cancer cells and normal cells was verified.Then,RBP4 was overexpressed in the two types of cells to detect the effect of RBP4 on their proliferation,migration and invasion,and to study the possible mechanism of its action.Methods:1.The differential expression of RBP4 in BEAS-2B and A549 cells was verified by RT-qPCR and Western Blot.2.The RBP4 plasmid was transfected into BEAS-2B and A549 cells.CCK-8,EDU and clone formation assay were used to detect the effect of overexpression of RBP4 on proliferation of BEAS-2B and A549 cells.The effect of overexpression of RBP4 on migration of BEAS-2B and A549 cells was detected by scratch assay.Transwell assay was used to detect the effect of overexpression of RBP4 on invasion of BEAS-2B and A549 cells.The effects of overexpression of RBP4 on the protein levels of MMP2,MMP9,PI3 K and AKT in BEAS-2B and A549 cells were detected by Western Blot.The nude mouse model was constructed to detect the effect of overexpression of RBP4 on tumor growth.3.RBP4 inhibitor A1120 was used to detect the effect of A1120 on A549 cell proliferation by CCK-8,EDU and clone formation assay.The effect of A1120 on the migration of A549 cells was detected by scratch test.Effect of Al120 on invasion of A549 cells by Transwell assay.Western Blot was used to detect the effects of A1120 on the levels of MMP2,MMP9,PI3 K and AKT in A549 cells.Results:1.RT-qPCR results showed that the RBP4 gene level in A549 cells was higher than that in BEAS-2B cells.Western Blot results proved that RBP4 protein level in A549 cells was higher than that in BEAS-2B cells.2.After overexpression of RBP4,CCK-8,EDU and clone formation results displayed that the proliferation ability of BEAS-2b and A549 cells was enhanced.The results of scratch assay showed that RBP4 promoted migration of BEAS-2B and A549 cells.Transwell results indicated that RBP4 promoted invasion of BEAS-2B and A549 cells.Western Blot results revealed that RBP4 up-regulated the expression of MMP2,MMP9,PI3 K and AKT in BEAS-2B and A549 cells.And overexpression RBP4 promoted tumor growth in vivo.3.CCK-8,EdU and colony formation results showed that A1120 significantly inhibited the proliferation of A549 cells.Scratch and Transwell assay confirmed that the inhibitor A1120 inhibited the migration and invasion of A549 cells.Western Blot results indicated that the inhibitor A1120 inhibited the expression of MMP2,MMP9,PI3 K and AKT in A549 cells.Conclusion:1.Compared with human normal lung epithelial cells BEAS-2B,RBP4 was highly expressed in human non-small cell lung cancer cell A549.2.Overexpression of RBP4 promoted proliferation,migration and invasion of BEAS-2B and A549 cells,and in vivo experiments proved that RBP4 promoted tumor growth.3.RBP4 inhibitor A1120 inhibited the proliferation,migration and invasion of A549 cells.4.RBP4 regulated A549 cells through PI3K/ AKT signaling pathway.