The Suppressive Effect Of MicroRNA-1on The Growth Of Xuanwei Lung Adenocarcinoma In Vitro

Background：XuanWei is famous of its highest incidence and mortality with lungcancer in China, Female mortality ranks higher incidence than any other area inChina,however,the pathogenesis of XuanWei lung cancer has been poorlyilluminated.microRNAs are a class of non-coding small RNAs that play importantroles in tumor. Recent researches have shown that microRNAs exert their tumorsuppressor or oncogenic function by regulate the expression of its target genes. It hasbeen suggested that microRNA expression profiles are differently between lungcancer and normal tissue. In the present study, we identified that compared toKMB-17,the expression level of microRNA-1is down-regulated in XuanWei lungcancer cells and the over-expression of microRNA-1can significantly inhibit thegrowth of XuanWei lung cancer cell by targeting the expression of c-met, a receptortyrosine kinase.Our finding revealed that microRNA-1acts a tumor suppressor inXuanWei lung cancer by repressing the specific proto-oncogene named c-met.Wealso identified that c-met is one of target proteins of microRNA-1.Thus, microRNA-1maybe a new access to the treatment for XuanWei lung cancer.Objective: To explore the suppressive effect of microRNA-1on the growth ofXuanWei lung cancer cell and regulatory effect on the expression of c-met.Methods:1.The molecular pre-microRNA-1sequence was synthesized by a chemicalmethod and cloned on the vector PCMV-MIR. then restrict the eukaryotic expressionplasmid PCMV-premiR-1.The constructed recombinant plasmid was identified by restriction enzyme digestion and sequence analysis.XuanWei lung cancer cell wascultured in vitro.Both blank plasmid and the recombinant eukaryotic expressionplasmid PCMV-premiR-1were transfected to XuanWei lung cancer cell bylipofectamine2000reagant.2. The effect of transfect ion was directly observed underfluorescence microscope and expression of microRNA-1in each group was tested byqPT-PCR.After transcript, the level of c-met protein was tested by western blot.3.The proliferation of XuanWei lung cell was detected by MTT assay. The ratio ofapoptosis in each group was analysis by flow cytometry.Results: the result of restriction end nuclease cleaved and DNA sequenced indicatedthat the recombinant plasmid was constructed successfully and fragment of premiR-1was correctly. Results of qRT-PCR indicated that compared with un-transfected groupand blank plasmid.The expression level of microRNA-1was significantlyup-regulated in PCMV-premiR-1transfected group.(p<0.05).Over expression ofmicroRNA-1can inhibit the growth of XuanWei lung cancer cell. Both the proteinand mRNA level of c-met are decreased in XuanWei lung cancer cell that tranfectedwith PCMV-premiR-1recombinant plasmid.Conclusions：microRNA could significant suppress the growth of XuanWei lungcancer, which closely related to down-regulate the expression of c-met...