Research Of MicroRNA-222-3p Related With Invasion And Metastasis In The Lacrimal Gland Adenoid Cystic Carcinoma

?Object?Of the malignant epithelial tumors,Adenoid cystic carcinoma(ACC)is the most common.ACC of the lacrimal gland is a challenging disease to manage due to a high recurrence rate,propensity for intracranial invasion,and poor prognosis.In order to solve this problem,many studies have been carried out.In recent years,it has been found that microRNA is involved in the occurrence and development of tumors,and it can play a role of oncogenes or antioncogene by regulating the expression of its target genes.Our aim is to determine whether miR-222-3p clusters regulate the biological processes of adenoid cystic carcinoma cells.At the same time,we identified the potential target gene of miR-222-3p and further elucidated the molecular mechanism of miR-222-3p in adenoid cystic carcinoma of the orbital lacrimal gland.? Methods ? we detect miR-222-3p expression in the cell lines of adenoid cystic carcinoma by real-time RT-PCR assay.Ability of ACC cell migration,invasion and proliferation was examined by MTT assay,transwell,clone formation assay.ACC cells with miR-222-3p overexpression were subjected to qRT-PCR and western blot for epithelial-mesenchymal transition(EMT)markers levels.TargetScan was used to predict putative targets of miR-222-3p.Then,the examination was performed whether miR-222-3p targets ARNT by the use of fluorescence reporter assays and western blotting.As target gene of microRNA-222,real-time quantitative PCR and Western blot results showed that after overexpression or blocking of microRNA-222 in adenoid cystic carcinoma cells,the expression level of ARNT mRNA and protein decreases or increases.Overexpression of ARNT in adenoid cystic carcinoma cells reduces cell proliferation,migration and invasion,and inhibits EMT process.Rescue experiments demonstrated that exogenous ARNT could partially counteract the effect of microRNA-222 on the function of adenoid cystic carcinoma cells.?Results? Real-time quantitative PCR detection show that miR-222-3p in ACC-M was upregulated compared with ACC-2 cells.After over-expression of microRNA-222 in adenoid cystic carcinoma cells,cell proliferation,migration and invasion were enhanced,and EMT process was promoted.The expression ofE-cadherin in epithelial cells was decreased,and the expression of Vimentin and ICAM-1 in interstitial cells was increased.The results of fluorescence reporter vector experiment showed that overexpression of miR-222-3p suppressed the expression of ARNT by directly binding to the 3? UTR of its mRNA.Real-time quantitative PCR and Western blot results showed that after over-expression or blocking of microRNA-222 in adenoid cystic carcinoma cells,the expression level of ARNT mRNA and protein decreased or increased accordingly.Overexpression of ARNT in adenoid cystic carcinoma cells reduces cell proliferation,migration and invasion,and inhibits EMT process.Rescue experiments demonstrated that exogenous ARNT could partially counteract the effect of microRNA-222 on the function of adenoid cystic carcinoma cells.?Conclusions?miR-222-3p can inhibit the expression of ARNT by targeting ARNT,thus promoting the proliferation,migration and invasion of adenoid cystic carcinoma cells,and causing interepithelial initialization.It plays a role similar to oncogene.It participates in the process of invasion and metastasis of adenoid cystic carcinoma of lacrimal gland.It provides a new field and idea for the research and treatment of adenoid cystic carcinoma of lacrimal gland.