Lenvatinib Inhibited Huh-7 Cell Migration And Invasion Via The UHRF1/DNMT1 Axis

Liver cancer is one of the most common and frequent malignant tumors in the world.The latest figures show that China ranks fifth in the incidence of liver cancer and second in the death rate.Metastatic recurrence is the main cause of death for all types of tumors,so it is urgent to find effective methods to inhibit the recurrence and metastasis of liver cancer.Lenvatinib,an oral multitarget tyrosine kinase inhibitor,was approved in 2018 as a first-line treatment for advanced liver cancer.Since the use of sorafenib,the efficacy is not inferior to sorafenib,but there are still few studies on the mechanism of action of sorafenib treatment,especially in the recurrence and metastasis of liver cancer.In this study,we investigated the effects of Lenvatinib on invasion and metastasis of Huh-7 cells and its related molecular mechanisms.We first examined the toxic effects of Lenvatinib on Huh-7 cells,and found that Lenvatinib could inhibit the epithelial-mesenchymal transformation(EMT)of Huh-7 cells.Lenvatinib can inhibit HUH7 invasion and metastasis in vivo and in vivo by Transwell assay and mouse caudal vein injection model.In addition,Lenvatinib can inhibit the extracellular and extracellular adhesion of Huh-7 cells,induce cytoskeleton recombination and inhibit pseudopod formation.E-cadherin plays a key role in the process of epithelial mesenchymal transformation(EMT),and EMT is closely related to tumor metastasis.Lenvatinib can positively regulate the expression level of E-cadherin.Through methylation specific PCR(MSP),we found that Lenvatinib can reduce the methylation level of E-cadherin promoter.The expression level of DNMT1 and UHRF1 was closely related to the methylation level of E-cadherin.The experiment showed that Lenvatinib could inhibit the expression of DNMT1 and UHRF1.The recovery experiment showed that Lenvatinib mediated the expression of E-cadherin through DNMT1 and UHRF1.Further molecular mechanism studies showed that Lenvatinib down-regulated DNMT1 and UHRF1 through ubiquitination pathway and reduced half-life.In conclusion,our results suggest that Lenvatinib regulates E-cadherin expression through the UHRF1/DNMT1 axis,thereby affecting Huh-7 cell migration and invasion.Our results reveal for the first time that Lenvatinib inhibits Huh7 cell invasion and metastasis and the specific mechanism of action.It is suggested that targeting DNMT1 and UHRF1 at the same time may be a new strategy for the treatment of HCC.