MiR-212 Facilitates The Migration And Invasion Of Esophageal Squamous Cell Carcinoma

BackgroundEsophageal squamous cell carcinoma(ESCC)patients with the same TNM stage often experience significantly different outcomes due to early metastasis happening or not.We previously found that miR-212 was significantly up-regulated in ESCC patients with poor outcome due to early metastasis,implying that miR-212 might promote ESCC metastasis.In order to explore the metastasis susceptibility of esophageal carcinoma from the cellular and molecular level,we will overexpress miR-212 in ESCC cell lines by transfecting miR-212 lentivirus,and then the effects of miR-212 on cell proliferation and migration will be detected by MTT and Transwell assay.This study revealed that miR-212 promoted ESCC cell migration and invasion rather than proliferation.This effect of miR-212 might be driven by promotion of ESCC cell EMT and production of ECM-degrading enzymes.Berberine treatment inhibited miR-212-mediated ESCC cell migration via blocking EMT.Thus,miR-212 might be as a prognosis marker and therapeutic target for ESCC,and berberine could be useful as a potential therapeutic drug against metastasis of ESCC patients with high miR-212 expression.ObjectiveTo explore and elucidate the effect and possible mechanism of miR-212 on esophageal squamous cell carcinoma cells.And the results will provide a new biomarker and strategy for ESCC metastasis precision diagnosis and therapy.MethodsThree cell lines of ESCC,namely KYSE-450,TE-1 and Eca109 were used.Lentivirus-mediated miR-212 was overexpressed in ESCC cells.The effect of miR-212 on cell proliferation was evaluated by MTT assay.Cell motility and invasiveness were determined by using transwell chambers without coating and with coating matrigel.Western blot was performed to detect the expression level of proteins related to epithelial-mesenchymal transition(EMT).Western blot were performed to test proteins level of extracellular matrix degrading enzymes,MMP2,MMP9 and u PA.Furthermore,we detected some potential therapeutic drugs and inhibitors which could inhibite the promoting role of miR-212 on ESCC cell migration.ResultsOverexpression of miR-212 had no obvious effect on ESCC cell proliferation,but significantly promoted migration and invasion of ESCC cells.Western blots showed that miR-212 decreased E-cadherin levels and increased the expression of ?-catenin,Vimentin as well as transcription factor Twist1.Western blot results showed that miR-212 augmented protein expression of MMP9 and u PA in a cell specific manner.Even though multiple chemicals and drugs,which display anti-tumor functions in many other types of cancer cells,had no effect on miR-212 induced cell phenotype in ESCC cells,berberine clearly rescued miR-212-induced cell migration phenotype and exhibited an inhibitory effect on EMT in ESCC cells.ConclusionsmiR-212 facilitated ESCC motility and invasiveness by promoting the EMT process as well as production of ECM-degrading enzymes,and berberine had an inhibitory effect on miR-212-induced ESCC cell migration.Collectively,miR-212 might be useful as a prognosis marker and therapeutic target for ESCC patients,and berberine might be a potential therapeutic drug against metastasis of ESCC patients with high miR-212 level.