Abstract
Objective To investigate the effect of silencing RET gene expression on IT (human medullary thyroid carcinoma cell lines) cell proliferation and its mechanism.Methods The double strands DNA containing RET gene was cloned into the pSilencerTM 3,1-H1 neo eukaryotic expression vector,and then was transfected into IT cells expressing high level of RET by lipofactamine.The levels of RET mRNA and protein expression and extracellular signal regulated kinase (ERK) phosphorylation were detected by real-time PCR and Western blotting.TT cell proliferation was tested by cell counting kit-8.Results The recombinant plasmid was successfully constructed as shown by sequencing.After the recombinant plasmid was transfected into TT cells,ret mRNA and RET protein expressions were reduced,along with decreased TT cell proliferation and ERK phosphorylation (P＜0.05).Conclusions The pSilencer-ret plasmid decreases the level of RET expression and inhibits TT cell proliferation,which is involved in ERK pathway.This may provide a new gene therapy approach for medullary thyroid carcinoma. Key words: RNA interfering; Ret gene; Medullary thyroid carcinoma; Gene therapy
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