Study On Mechanism Of CCDC85B Promotes Non-small-cell Lung Cancer Cell Proliferation And Invasion

Lung cancer is one of the leading causes of death in malignant tumors.Although lung cancer treatment has made great progress in recent years,complicated factors play an important role in invasion and metastasis.Survival rate is still not high.Therefore,to clarify the mechanism of invasion and metastasis is an important task of prevention and treatment of cancer.Wnt/?-catenin signaling pathway plays an important role in the early embryonic development,adult tissue steady-state maintenance,stem cell regulation and tumorigenesis.The core signal transduction factor ?-catenin,which is activated by the activation of the canonical Wnt signaling pathway,CK1?/? actived,the actived CK1?/? phosphorylates Dishevelled(Dvls),eventually leading to cytoplasmic ?-catenin not being ubiquitinated,However,?-catenin which accumulates and enters to the nucleus,binds to Tcf-4 and activates the transcriptional activity of target genes such as c-myc,Cyclin D1,MMP7 and promote the proliferation,invasion and metastasis of tumor.There are many families of CCDC related domain family members.Previous studies have reported that p53 can induce CCDC85 B expression,CCDC85 B interacts with TCF4 in the nucleus,and CCDC85 B can down-regulate the canonical Wnt signaling pathway by competeing with ?-catenin in combination with TCF4 transcription factor.However,the expression of CCDC85 B in lung cancer and the proliferation and invasion of lung cancer cells have no report,Which arouse our great interest.Objective: In this study,we detected the expression of CCDC85 B in non-small cell lung cancer and lung cancer cells and its relationship with clinicopathological factors,and to expore the mechanisms and influences of CCDC85 B in proliferation and invasion of non-small cell lung cancer.Methods: Western blot and immunohistochemistry were used to investigate theexpression of CCDC85 B protein in non-small cell lung cancer cell lines and tissues,and its relationship with clinicopathological factors.The expression of CCDC85 B in lung cancer cell line was studied by Western blot.Colony formation,MTT and Transwell were used to study the effect of CCDC85 B on the proliferation and invasion of non-small cell lung cancer.Results: 1.In lung cancer tissue,the expression of CCDC85 B in the cytoplasm,which is higher than that in adjacent normal lung tissue.The high expression of cytoplasm of CCDC85 B is positively correlated with high TNM stage and lymph node metastasis in lung cancer patients.Western blot results donate that in freshly matched lung cancer tissues the expression of CCDC85 B were significantly higher than those of adjacent tissues.The expression of CCDC85 B in lung cancer cell lines was higher than that in normal bronchial epithelial cell line HBE.2.In A549 and H1299 lung cancer cell lines,overexpression of CCDC85 B promoted the cell proliferation,colony formation and invasiveness,while si RNA-mediated CCDC85 B knockdown exhibited the opposite effects.3.CCDC85 B upregulates Wnt signaling pathway acticity.In A549 and H1299 cells,CCDC85 B overexpression downregulated p-?-catenin expression,upregulated active ?-catenin and the downstream factors: c-myc,,cyclin D1 and MMP-7 protein expression.Conversely,CCDC85 B knockdown upregulated p-?-catenin expression,downregulated active ?-catenin and the downstream factors: c-myc,cyclin D1 and MMP-7 protein expression.4.CCDC85 B through activation of PI3K/AKT/GSK3? pathway regulates WNT signaling pathway activity.In A549 and H1299 cells CCDC85 B overexpression upregulated p-AKT Ser 473,p-GSK3?,and CCDC85 B knockdown downregulated p-AKT Ser 473,p-GSK3?.We treated CCDC85B-overexpressing A549 and H1299 cells with the AKT inhibitor,LY294002(10 ?M).We discovered that p-?-catenin upregulated and active-?-catenin,p-GSK3?downregulated.Conclusion: 1.In lung cancer tissue,the expression of CCDC85 B in the cytoplasm.CCDC85B overexpression is positively correlated with high TNM staging and lymph node metastasis in lung cancer patients.2.CCDC85 B promoted the proliferation and invasion of non-small cell lung cancer.3.CCDC85 B through activation of PI3K/AKT/GSK3? pathway upregulates WNT signaling pathway activity promotes the proliferation and invasion of lung cancer cell.