WWP2 Promotes The Proliferation And Metastasis Of Lung Cancer Cells Through Hippo Pathway And WNT Pathway

Objective:It has been reported that WWP2,a member of HECT E3 ubiquitase family,plays an important role in the development of a variety of cancers,while lung cancer still accounts for a large proportion of cancer deaths.Moreover,WWP2 has 4 WW domains,while Hippo signaling pathway is rich in WW domains,and WNT signaling pathway is in series with Hippo signaling pathway.Therefore,the effect mechanism of WWP2 on cancer through Hippo and WNT signaling pathways was explored in non-small cell lung cancer.Research methods:Database and immunohistochemistry were used to analyze the relationship between WWP2 and clinicopathological factors in patients with non-small cell lung cancer.WWP2 expression was detected in non-small cell lung cancer cell lines,and the location of WWP2 was detected by immunofluorescence.After transfection and interference with non-small cell lung cancer cells,The effects of WWP2 on the proliferation and migration ability of non-small cell lung cancer were verified by cell functional experiments,and then the effects of WWP2 on Hippo and WNT signaling pathway activities and EMT were detected by Western Blot and RT-PCR.Results: 1.The expression level of WWP2 was related to clinicopathological factors in NSCLLC patients.Immunohistochemical results showed that WWP2 was highly expressed in NSCLC tissues,and its high expression was positively correlated with TNM staging of NSCLC and lymph node metastasis,and was closely correlated with poor prognosis of patients with NSCLC.Immunofluorescence mapping results showed that WWP2 was expressed in both cytoplasm and nucleus of NSCLC,but mainly in the nucleus.2.High expression of WWP2 can promote the proliferation and migration of lung cancer cells.The expression of WWP2 in NSCLC cell lines was detected by western blot assay.Three cell lines,A549,H1299 and H1975,were selected for transfection and interference experiments.We found that transfection with exogenous WWP2 could promote the proliferation and migration of NSCLC cells.Interference with endogenous WWP2 can inhibit the proliferation and migration of NSCLC.3.WWP2 can affect the activity of Hippo and WNT signaling pathways.It has been reported that the WW domain can bind to Merlin,which was proved by immunocoprecipitate experiment,and then western blot experiment verified that Hippo pathway can be inhibited and WNT signaling pathway activity can be promoted when WWP2 expression is high,and opposite results can be obtained when WWP2 expression is low.4.WWP2 is related to EMT.western blot experiment showed that WWP2 could promote EMT process when its expression was high,but the opposite result was obtained when its expression was low.Conclusion: WWP2 affects the activity of Hippo and WNT signaling pathways by binding to Merlin,thus regulating the proliferation and migration of non-small cell lung cancer.