Citation

  • Authors: Wang, H., Cao, F., Li, X., Miao, H., E, J., Xing, J., Fu, C. G.
  • Year: 2015
  • Journal: BMC Cancer 15 748
  • Applications: in vitro / mimic miRNA, mimic miRNA and DNA cotransfection / INTERFERin, jetPRIME
  • Cell types:
    1. Name: HCT 116
      Description: Human colon carcinoma cells
      Known as: HCT116
    2. Name: HEK-293
      Description: Human embryonic kidney Fibroblast
      Known as: HEK293, 293
    3. Name: SW480
      Description: Human colon adenocarcinoma cells

Method

50 nM of siRNA. Co-transfection of 50nM miRNA mimics with 13 ng DNA in 96-well plates.

Abstract

BACKGROUND: MicroRNAs (miRNAs) are small noncoding RNAs that potentially play a critical role in tumorigenesis. Mounting evidence indicates that one specific miRNA: miR-320b is down regulated in numerous human cancers, including colorectal cancer (CRC); making the hypothesis that miR-320b may play a key role in tumorigenesis plausible. However, its role in carcinogenesis remains poorly defined. The goal of this study is to better clarify the role of miR-320b in tumor growth of CRC. METHODS: Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was conducted to detect the expression of miR-320b in CRC tissues and 5 CRC cell lines. The effect of miR-320b on cell proliferation was analyzed in vitro and in vivo. Furthermore, a luciferase reporter assay was performed to measure the target effects of miR-320b. Lastly, the messenger RNA (mRNA) and protein levels of the gene c-MYC were measured in CRC cell lines and tissues by qRT-PCR, and confirmed via Western blot and Immunohistochemical (IHC) staining. RESULTS: The results presented here showed that miR-320b expression was down regulated in both CRC tissues and cells. Overexpression of miR-320b in CRC cells was statistically correlated with a decrease of cell growth in vitro and in vivo, while c-MYC was identified as a target gene of miR-320b in CRC. Furthermore, it was found that up-regulation of c-Myc can attenuate the effects induced by miR-320b. CONCLUSIONS: Our identification of c-MYC as a target gene of miR-320b provides new insights into the pathophysiology of CRC proliferation, and identifies miR-320b as a novel therapeutic target for the treatment of CRC.

Pubmed