SPP1, analyzed by bioinformatics methods, promotes the metastasis in colorectal cancer by activating EMT pathway

Biomed Pharmacother. 2017 Jul:91:1167-1177. doi: 10.1016/j.biopha.2017.05.056. Epub 2017 May 17.

Abstract

Objective: Tumor metastasis is still a great challenge for the prognosis of colorectal cancer (CRC). Although secreted phosphoprotein 1 (SPP1) over-expression is confirmed to associate with invasion, metastasis of CRC, the underlying mechanism by which modulates the CRC metastasis is still not fully explained.

Method: GDS4382 was obtained from GEO database and differentially expressed genes (DEGs) were analyzed by bioinformatics methods 55 paired samples of CRC and adjacent non-cancerous tissues were collected to detect the expression of SPP1 by q-PCR and western blot. Functional analysis of siRNA-SPP1, including proliferation, apoptosis, colony formation, cell cycle, migration, was investigated in CRC cell lines and tumor xenografts were conducted in nude mice. Protein expression of E-cadherin and vimentin was detected by western blot.

Results: 1887 DEGs were analyzed and selected from GDS4382, of which, SPP1 and epithelial-mesenchymal-transition (EMT) showed a close association by bioinformatics analysis. The mRNA and protein expression of SPP1 were significantly higher in CRC tissues than that in adjacent non-cancerous tissues (P<0.05). Overexpression of SPP1 closely associated with tumor invasion, metastasis and low survival in CRC. Moreover, siRNA-SPP1 repressed proliferation, cell cycle, colony formation, migration and tumor growth in vivo and promoted cell apoptosis in CRC cell lines. In addition, Protein expression of E-cadherin was obviously up-regulated and Vimentin was down-regulated in CRC cells after siRNA-SPP1 (P<0.05).

Conclusion: SPP1 expression was significantly up-regulated in CRC. And SPP1 promoted the metastasis of CRC by activating EMT, which could be a potentially therapeutic target for patients with CRC.

Keywords: Bioinformatics; Colorectal cancer; Epithelial-mesenchymal-transition; Metastasis; Secreted phosphoprotein 1.

MeSH terms

  • Animals
  • Apoptosis / genetics
  • Cadherins / genetics
  • Cell Cycle / genetics
  • Cell Line, Tumor
  • Cell Movement / genetics
  • Cell Proliferation / genetics
  • Colorectal Neoplasms / genetics*
  • Colorectal Neoplasms / pathology
  • Computational Biology / methods
  • Down-Regulation / genetics
  • Epithelial-Mesenchymal Transition / genetics*
  • HCT116 Cells
  • Humans
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Osteopontin / genetics*
  • Prognosis
  • RNA, Messenger / genetics
  • RNA, Small Interfering / genetics
  • Up-Regulation / genetics
  • Vimentin / genetics

Substances

  • Cadherins
  • RNA, Messenger
  • RNA, Small Interfering
  • SPP1 protein, human
  • Vimentin
  • Osteopontin