MicroRNA-223 functions as an oncogene in human gastric cancer by targeting FBXW7/hCdc4

  • Jinhai Li
  • Yuanyuan Guo
  • Xiaodi Liang
  • Ming Sun
  • Guoliang Wang
  • Wei De
  • Wenxi WuEmail author
Original Paper



The aim of this study was (a) to determine the role of micro-223 (miR-223) in gastric cancer and (b) to elucidate its regulatory mechanism on the FBXW7/hCdc4 gene.

Materials and methods

Artificial miR-223 and control oligonucleotide was transfected into gastric cancer cell line SGC7901 by using Lipofectamine2000. Apoptosis of miR-223 group and control group cells was analyzed by flow cytometry, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide and colony formation assays were performed to detect the cell viability, to survey migration of miR-223 group and control group cells; scratch wound-healing motility assays, Transwell Assay, and Western blot test were performed to measure the variance of hFBXW7. Luciferase Reporter Assay, which was done by pLUC-hFBXW7 WT-3′-UTR co-transfected with pLMP-hsa-miR-223 or pLMP plasmid (as control) into HEK293T cells, used to detect whether hFBXW7 is a direct target gene of miR-223. Gastric cancer cell line SGC7901 transfected with miR-223 or control oligonucleotide was resuspended in ECM gel and then was injected into the flank of nude mice, 4 weeks later, the nude mice were euthanized. The tumors were excised then were measured and weighted. SYBR-Green I-based real-time RT-PCR study was used to detect the level of miR-223 in 22 gastric cancer tissue and corresponding gastric mucosa tissues. Immunohistochemical method was applied to detect the protein of hFBXW7.


Gastric cancer cell line SGC7901, transfected with miR-223, showed significant reduction in cellular apoptosis and increased proliferation and invasion in vitro. Similar results were found in tumorigenesis assays performed in nude mice. Moreover, 19 of 22 cancer tissue samples highly expressed miR-223, when compared with patient-matched normal gastric mucosa. Specifically, patients with lymph node metastasis or metastatic disease (M1) at an advanced pathological stage showed significantly higher expression of miR-223. FBXW7/hCdc4 protein (FBW7) levels in gastric cancer cases were inversely correlated with miR-223 expression. Overexpression of miR-223 in gastric cancer cell lines decreased FBW7 expression at the translational level and decreased FBXW7/hCdc4-driven luciferase-reporter activity.


In summary, the data indicated that miR-223 targets FBXW7/hCdc4 expression at the post-transcriptional level and appears to regulate cellular apoptosis, proliferation, and invasion in gastric cancer. MiR-223 may serve as a novel therapeutic target in gastric cancer.


MicroRNAs MiR-223 Proliferation FBXW7/hCdc4 Gastric cancer 



We thank Dr. Bin Cao and Lizong Shen of Jiangsu province hospital for helpful. This work was supported by National Science Foundation of China (No. 81070381 and No. 30872532).

Conflict of interest

We declare that we have no conflict of interest.

Supplementary material

432_2012_1154_MOESM1_ESM.tif (6.6 mb)
Fig. 1 Sample QC report (A) Total SGCS RNA sample, the ratio of 28/18 s was 1.9. (B) The ratio for SGCS0 sample was 2.1. They were validated samples and used for additional experiments (TIFF 6783 kb)
432_2012_1154_MOESM2_ESM.tif (2.2 mb)
Fig. 2 RT-PCR products using SYBR-Green I for miR-223. Amplicons were analyzed by 8% PAGE electrophoresis. MiR-223 bands at 60 bp position and GAPDH at 180 bp position indicated that the PCR products were as desired (G0, SGCG0; S, SGCS) (TIFF 2271 kb)
432_2012_1154_MOESM3_ESM.docx (39 kb)
Supplementary material 3 (DOCX 39 kb)


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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Jinhai Li
    • 1
  • Yuanyuan Guo
    • 2
  • Xiaodi Liang
    • 2
  • Ming Sun
    • 2
  • Guoliang Wang
    • 1
  • Wei De
    • 2
  • Wenxi Wu
    • 1
    Email author
  1. 1.Department of Gastrointestinal SurgeryFirst Affiliated Hospital of Nanjing Medical UniversityNanjingPeople’s Republic of China
  2. 2.Department of Biochemistry and Molecular BiologyNanjing Medical UniversityNanjingPeople’s Republic of China

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