The Role Of ANKHD1 On Proliferation,migration And Invasion Of Colorectal Cancer And Its Mechanism

Objective:The expression of ANKHDI was detected in colorectal cancer specimens,and the relationship between ANKHD1 expression and clinicopathological features of colorectal cancer patients was analyzed.To investigate the effect of ANKHDI on the biological function of colorectal cancer.To study the role of YAP1 in the progression of colorectal cancer influencedby ANKHD1.Methods:ANKHD1 expression was detected by immunohistochemistry(IHC)in 136 colorectal cancer tissues.We analyzed the relationship between the expression of ANKHD1 and clinicopathological features of patients.Cell lines those stably down-regulating ANKHD1 expression were constructed by lentiviral infection.CCK8 assay,Transwell assay and flow cytometry were used to detect the proliferation,invasion and metastasis ability and apoptosis of colorectal cancer cells after ANKHD1 silencing.Western blot and real-time PCR were used to detect the expression of YAP1 and EMT-related proteins after ANKHD1 silencing.Using balb/c mice to inoculate tumors by subcutaneous tumor-bearing method,observe the effect of ANKHD1 silencing on tumor growth in vivo.The tumor metastasis model was established by using balb/c mice.The experiment was terminated at 6 weeks.The main organs of the mice were taken out.The tumor metastasis was observed by macroscopic observation and HE staining,and the effect of ANKHDI on tumor invasion and metastasis ability was further studied.The plasmid expressing YAP1 was transfected into ANKHD1-silencing cells by transient transfection,and the transfection efficiency was detected by Western blot.CCK8 assay,Transwell assay and Western blot assay were used to detect the proliferation,invasion and metastasis and the expression of EMT-related proteins in ANKHD1 silenced cell lines after overexpression of YAP1.Results:The expression of ANKHD1 in colorectal cancer was positively correlated with the depth of tumor invasion.The higher the expression of ANKHD1,the deeper the tumor infiltration,with statistical significance.Down-regulation of ANKHD1 inexpression in colorectal cancer cells HCT116 and SW480 inhibits cell proliferation,invasion and metastasis,but does not affect apoptosis.Silencing of ANKHD1 significantly inhibits the occurrence of EMT in colorectal cancer cells,inhibits the activation of AKT signaling and the activity of YAP1 in the Hippo signaling pathway.ANKHD1 silencing inhibits the growth of colorectal cancerinvivoand inhibits lung metastasis of the tumor.Overexpression of YAP1 reverses the inhibition of proliferation,invasion and migration of colorectal cancer cells by ANKHD1 silencing,and reverses the inhibition of EMT in colorectal cancer cells caused by ANKHD1 silencing.Conclusions:The expression of ANKHD1 in colorectal cancer is related to the depth of tumor invasion.ANKHD1 promotes the proliferation,invasion and metastasis of colorectal cancer cells.ANKHD1 regulates the production of EMT and the activation of AKT signaling in colorectal cancer cells by targeting YAP1.