Silencing effect of lentivirus-mediated RNA interference on metastasis-associated gene 1(MTA1) in human nasopharyngeal carcinoma cell line / 解放军医学杂志

ABSTRACT

Objective To investigate the difference between two substrains,5-8F and 6-10B,of nasopharyngeal carcinoma cell line SUNE1on expressing metastasis-associated gene 1 (MTA1),and evaluate the silencing effect of lentivirus-mediated RNA interference (RNAi) on MTA1 in human nasopharyngeal carcinoma cell line.Methods The differential expression of MTA1 mRNA and MTA1 protein in 5-8F and 6-10B cell lines were detected by real time PCR and Western blotting respectively.Short interference RNA (siRNA) fragment targeting MTA1 gene was designed using online databases and software.A specific RNAi lentiviral vector targeting human MTA1 gene was constructed and transfected into nasopharyngeal carcinoma cell line 5-8F.Real time PCR and Western blotting were performed to detect the expressive changes in MTA1 mRNA and MTA1 protein in the transfected 5-8F cells respectively.Results Compared with cell line 6-10B,the expressions of MTA1 mRNA and MTA1 protein were higher in 5-8F cell line.Sequence analysis validated the correct insert of siRNA targeting MTA1 gene into the lentiviral vector.Real time PCR and Western blotting results showed that the expressions of MTA1 mRNA and MTA1 protein in 5-8F cell lines were down-regulated significantly after siRNA transfection.Conclusions MTA1 may promote the malignant transformation and enhance the metastatic potential of nasopharyngeal carcinoma cell lines.The expression of MTA1 in 5-8F cells can be inhibited effectively by MTA1-specific siRNA expression lentiviral vector,which provides a valuable tool for investigating the role of MTA1 gene in the carcinogenesis and progression of nasopharyngeal carcinoma.