Effect Of YB-1 Downregulation Via RNAi On MACC1 Gene Expression In Human Lung Adenocarcinoma Cell Line A549

Objective:In recent years, the incidence of lung adenocarcinoma is on the rise,to more than 40% of the number of non-small-cell lung carcinoma (NSCLC).The clinical procedures is difficult for the early stage of metastasis. Metastasis-Associated in Colon Cancer 1(MACC1) is a new-found gene which has a close relationship with proliferation and metastasis of the tumor,and becoming one of the most important discovery in metastatic reaserch.Human Y box binding protein-1(YB-1) is an important transcription and translation factor which specific binds to the target gene promoter and enhancer with the Y-box sequences. Metastasis-Associated in Colon Cancer 1,MACC1)promtor region contains several Y-box sequence.To research the function of YB-1 protein could reveal the regulational mechanism of MACC1 expression.Based on the early research,we plan to use lipidosomal transfection and RNA interference,inhibit YB-1 protein expression in lung adenocarcinoma cell line A549,investigate MACC1 expression differentiation,initially confirm YB-1 protein on the regulation of MACC1 expression.Methods:We used the transfection reagent Lipofectamine2000 and transfected YB-1 siRNA duplex which designed and synthesized by Santa Cruz company into A549 lung adenocarcinoma cells,then the control siRNA group, Lipofectamine2000 group and normal group as control.Then using RT-PCR,Western Blot analysis of YB-1 and MACC1 gene and protein in each group in A549 cell line.The experimental data analysis by SPSS17.0 statistical software.Experiments of all groups will be repeated three times,the results of measurement data represent by mean±standard error(x±S),and using the single factor variance analysis whether the differences between experimental groups is significant(P<0.05). Result:1.The results of RT-PCR showed that after YB-1 siRNA interference,YB-1 mRNA expression downregulated(P<0.05).2. The results of Western Blot showed that after YB-1 siRNA interference,YB-1 protein expression also decreased(P<0.05),initially confirmed that YB-1 can regulate the expression of MACC1 gene in A549 cell line.Conclusion:The results of RT-PCR and Western Blot initial confirm that YB-1 regulates the expression of MACC1 gene.