Periostin in Angiogenesis and Inflammation in CRC-A Preliminary Observational Study

Abstract

Background and Objectives: To assess the periostin level and the concentrations of pro-inflammatory cytokines: TNFα, IFN-γ, IL-1β and IL-17 in tumor and marginal tissues of CRC and to investigate the influence of periostin on angiogenesis by MVD (microvessel density) and concentration of VEGF-A in relation to clinicopathological parameters of patients. Materials and Methods: The study used 47 samples of tumor and margin tissues derived from CRC patients. To determinate the concentration of periostin, VEGF-A, TNFα, IFNγ, IL-1β and IL-17, we used the commercially available enzyme- linked immunosorbent assay kit. MVD was assessed on CD34-stained specimens. The MVD and budding were assessed using a light microscope Results: We found significantly higher concentrations of periostin, VEGF-A, IFN-γ, IL-1 β, IL-17 and TNFα in the tumor samples compared with surgical tissue margins. The tumor concentrations of periostin were correlated with tumor levels of VEGF-A, IFN-γ, IL-1β and TNFα. We observed significant correlation between margin periostin and VEGF-A, IFN-γ, IL-17 and TNFα in tumor and margin specimens. Additionally, we found a significantly negative correlation between periostin tumor concentration and microvessel density at the invasive front. Tumor periostin levels were also correlated positively with tumor budding. Conclusions: Periostin activity may be associated with pro-inflammatory cytokine levels: TNFα, IFN-γ, IL-1β and IL-17. Our results also suggest the role of periostin in angiogenesis in CRC and its upregulation in poorly vascularized tumors. Further research on the regulations between periostin and cytokines are necessary to understand the interactions between tumor and immune tumor microenvironment, which could be helpful in the development of new targeted therapy.

Keywords: colorectal cancer; periostin; tumor microenvironment.

Figure 1

Box-plot- levels of periostin, VEGF-A, IFN-γ, IL-1 β, IL-17, TNFα molecules in tumor and tissue margins; protein levels are presented as log-transformed values as g/g. Paired T-student’s test.

Figure 2

Correlations between the levels of the examined molecules presented as a heatmap with hierarchic clustering. R—Pearson’s correlation coefficient, * p < 0.05.

Figure 3

CD34 immunostaining in CRC specimens used to assess microvessel density (A) 400× magnification, (B) 100× magnification.

Figure 4

Association between MVD at the invasive front and periostin concentration (R = −0.44, p = 0.036). R—Pearson’s correlation coefficient.

Figure 5

Budding assessment (200× magnification, Olympus BX 51 microscope, Olympus Life Science, Olympus manufacturer, Tokyo, Japan), (A) high budding tumor (6 buds in the camera field of view (FOV), more than 10 buds in full microscope FOV), (B) Low budding tumor, 1 bud in FOV.