Expression of ErbB3 in pancreatic cancer cell line regulated by microRNA-148a
10.3760/cma.j.issn.0254-1432.2013.06.009
- VernacularTitle:微小核糖核酸-148a调控胰腺癌细胞中ErbB3的表达
- Author:
Hui FENG
;
Weiyan YAO
;
Xi CHEN
;
Chenyu ZHANG
;
Yalei WANG
- Publication Type:Journal Article
- Keywords:
Pancreatic neoplasms;
microRNAs;
Oncogene proteins v-erbB;
Polymerase chain reaction;
Immunoblotting
- From:
Chinese Journal of Digestion
2013;(6):399-402
- CountryChina
- Language:Chinese
-
Abstract:
Objective To look for and confirm the downstream regulated gene of microRNA (miRNA)-148a in pancreatic cancer cell line.Methods The target gene regulated by miRNA-148a was predicted through bio-informatics analysis.The plasmid containing desired gene and with luciferase 3'-untranslated region (3'-UTR) reporter was constructed.Pancreatic cancer cells BXPC-3 were transfected with analogs and inhibitors of miRNA-148a by liposomes.The activity of luciferase was measured to determine whether miRNA-148a directly connected with desired gene.The expression level of miRNA-148a was changed in BXPC-3 cells,and the changes of target gene v-verb-b2 erythroblastic leukemia viral oncogene homolog 3 (awian) (ErbB3) expression were detected by Western blot at protein level.The data were analyzed by one way ANOVA.Results There was a conservative binding site of ErbB3 with miRNA-148a detected by bio-informatics analysis,miRNA-148a directly combined with ErbB3 and the activity of luciferase decreased to (25.00+47.00) ％ of the negative control (F=4.66,P＜ 0.01).After miRNA-148a overexpression,the gray value of ErbB3 expression in BXPC-3 cells decreased to (26.16±4.69)％ of control group (F=6.563,P＜0.05).Conclusion miRNA-148a directly targeted and regulated the expression of ErbB3 in pancreatic cancer cell line BXPC-3.
