Highly selective behaviour of colon adenoma after administration of EMR composition and its HCT116-based model.

When applying the improved composition of the solution used during endoscopic mucosal resection (EMR), we observed unexpectedly large and quantitatively significant differences in adenoma response vs. healthy tissue of the surrounding GIT tract, namely, the selective reaction enhancing the adenoma volume and differentiated colour. The in vitro experiments on the model neoplasia cell line HCT116 suggest that the robust differences in the response of starving cells can be traced down principally to tetrastarch digestion and the enhanced metabolic rate of neoplastic cells. The neoplastic tissue grows into several intestine layers so that submucosal injection of iso-oncotic tetrastarch compound leads to degradation of starch and production of oncotic molecules in submucosa transported by facilitated transport into the neoplastic tissue. The colour distinction is due to concentration differences of the reporting dye between three separated compartments, further enhancing the utility of the contrasting mixture. The diffusion dynamics shall be tuneable by optimizing starch composition, improving desirable pharmacokinetics.

Highly selective behaviour of colon adenoma after administration of EMR composition and its HCT116-based model.

When applying the improved composition of the solution used during endoscopic mucosal resection (EMR), we observed unexpectedly large and quantitatively significant differences in adenoma response vs. healthy tissue of the surrounding GIT tract, namely, the selective reaction enhancing the adenoma volume and differentiated colour. The in vitro experiments on the model neoplasia cell line HCT116 suggest that the robust differences in the response of starving cells can be traced down principally to tetrastarch digestion and the enhanced metabolic rate of neoplastic cells. The neoplastic tissue grows into several intestine layers so that submucosal injection of iso-oncotic tetrastarch compound leads to degradation of starch and production of oncotic molecules in submucosa transported by facilitated transport into the neoplastic tissue. The colour distinction is due to concentration differences of the reporting dye between three separated compartments, further enhancing the utility of the contrasting mixture. The diffusion dynamics shall be tuneable by optimizing starch composition, improving desirable pharmacokinetics.

Galectin-1 and Galectin-3 expression in colon cancer and its correlation with tumor invasion, differentiation, and metastatic spread

Aim. To study the expression of galectin-1 and galectin-3 in colon cancer and the levels of these proteins in the peripheral blood in relation to the differentiation, invasion, and metastatic dissemination.Materials and Methods. We examined primary tumors and the corresponding peripheral blood samples from 81 patients with colon cancer. Control group consisted of 49 patients with colon adenoma and 17 healthy volunteers. Expression of galectin-1 and galectin-3 in colon tissue was determined by immunohistochemical staining, while their plasma level was measured by enzyme-linked immunosorbent assay. Tumor staging was performed in accordance with the TNM system (AJCC, 2009). Cell differentiation was defined according to the respective clinical guidelines (Russian Cancer Association, 2018).Results. We detected an elevated expression of galectin-1 and galectin-3 in primary colon cancer as compared with colon adenoma and higher plasma levels of these proteins in colon cancer patients in comparison with healthy volunteers. High expression of tumor and plasma galectin-1 was associated with higher tumor stage (T3/T4) and the presence of local and distant metastases. Overexpression of galectin-3 in the primary tumor correlated with lower differentiation and lymph node metastasis.Conclusion. Galectin-1 and galectin-3 are involved in colon cancer progression and might be used as predictors of an adverse outcome.

High Variability in Cellular Proliferation, Gene Expression, and Cytokine Production in the Nonneoplastic Colonic Epithelium of Young Apc+/Min-FCCC Mice

An urgent need exists to identify efficacious therapeutic preventive interventions for individuals who are at high risk of developing colorectal cancer. To maximize the benefits of preventive intervention, it is vital to identify the time interval during which the initiation of a preventive intervention will lead to an optimal outcome. The goal of the present study was to determine if oncogenic events can be detected in the nonneoplastic colonic mucosa of Apc+/Min-FCCC mice prior to formation of the first adenoma, thus defining an earlier point of intervention along the cancer continuum. Tissues taken at three potential points of intervention were characterized: prior to Apc mutation (wild type Apc+/+-FCCC mice); after initiation but prior to colon adenoma formation (tumor-free Apc+/Min-FCCC mice); and after formation of the first colon adenoma (tumor-bearing Apc+/Min-FCCC mice). Experimentation focused on molecular processes that are dysregulated in early colon lesions: 1) cellular proliferation (proliferative index and size of the proliferative zone); 2) cellular stemness (expression of Ascl2, Grem1, Lgr5 and Muc2); 3) EGFR signaling (expression of Ereg); and 4) inflammation (expression of Mmp9, Ptsg2, and Reg4, as well as secretion of 18 cytokines involved in immune activation and response). Interestingly, the nonneoplastic colonic mucosa of wild type, tumor-free Apc+/Min-FCCC, and tumor-bearing Apc+/Min-FCCC mice did not display significant differences in average epithelial cell proliferation (fold change 0.8–1.3, p≥0.11), mucosal gene expression (fold change 0.8–1.4, p≥0.22), or secretion of specific cytokines from colonic mucosa (fold change 0.2–1.5, p≥0.06). However, the level of cytokine secretion was highly variable, with many (22% of wild type, 31% of tumor-free Apc+/Min-FCCC, and 31% of tumor-bearing Apc+/Min-FCCC) mice categorized as outliers (> 1.5 x interquartile ranges below the first quartile or above the third quartile) due to elevated expression of at least one cytokine. In summary, no differences were observed in proliferation, stemness, and EGFR signaling in the colonic mucosa of wild type vs Apc+/Min-FCCC mice, with low baseline cytokine expression, prior to the formation of the first colon adenoma. The results of this study provide valuable baseline data to inform the design of future cancer prevention studies.

Highly selective behaviour of colon adenoma after administration of EMR composition and its HCT116-based model.

When applying the improved composition of solution used during endoscopic mucosal resection (EMR), we have observed unexpectedly large and quantitatively significant differences in adenoma response vs. healthy tissue of surrounding GIT tract, namely the selective reaction enhancing the adenoma volume and differentiated colour. The in vitro experiments on model neoplasia cell line HCT116 suggest that the robust differences in the response of starving cells can be traced down principally to the tetrastarch digestion and enhanced metabolic rate of neoplastic cells. The neoplastic tissue grows into several intestine layers so that submucosal injection of iso-oncotic tetrastarch compound leads to degradation of starch and production of oncotic molecules in submucosa transported by facilitated transport into the neoplastic tissue. The colour distinction is due to concentration differences of the reporting dye between three separated compartments, further enhancing the utility of the contrasting mixture. The diffusion dynamics shall be tuneable by optimizing starch composition, improving desirable pharmacokinetics.

SPNS2 Downregulation Induces EMT and Promotes Colorectal Cancer Metastasis via Activating AKT Signaling Pathway

Spinster homologue 2 (SPNS2), a transporter of S1P (sphingosine-1-phosphate), has been reported to mediate immune response, vascular development, and pathologic processes of diseases such as cancer via S1P signaling pathways. However, its biological functions and expression profile in colorectal cancer (CRC) is elusive. In this study, we disclosed that SPNS2 expression, which was regulated by copy number variation and DNA methylation of its promoter, was dramatically upregulated in colon adenoma and CRC compared to normal tissues. However, its expression was lower in CRC than in colon adenoma, and low expression of SPN2 correlated with advanced T/M/N stage and poor prognosis in CRC. Ectopic expression of SPNS2 inhibited cell proliferation, migration, epithelial–mesenchymal transition (EMT), invasion, and metastasis in CRC cell lines, while silencing SPNS2 had the opposite effects. Meanwhile, measuring the intracellular and extracellular level of S1P after overexpression of SPNS2 pinpointed a S1P-independent model of SPNS2. Mechanically, SPNS2 led to PTEN upregulation and inactivation of Akt. Moreover, AKT inhibitor (MK2206) abrogated SPNS2 knockdown-induced promoting effects on the migration and invasion, while AKT activator (SC79) reversed the repression of migration and invasion by SPNS2 overexpression in CRC cells, confirming the pivotal role of AKT for SPNS2’s function. Collectively, our study demonstrated the suppressor role of SPNS2 during CRC metastasis, providing new insights into the pathology and molecular mechanisms of CRC progression.

ErbB Receptor Stimulation Is Required for Mouse Colon Adenoma Organoids to Form Crypts

The majority of colon adenomas harbor genetic mutations in the APC gene. APC mutation leads to changes in Wnt signalling and cell-cell adhesion: as a consequence, intestinal crypt budding increases and the excess crypts accumulate to form adenomas that progress to colon cancer. When cultured with Wnt, R-spondin, EGF, Noggin, myofibroblast conditioned medium and Matrigel, crypts from normal mouse colon mucosa form crypt-producing organoids and can be passaged continuously. Under the same culture and passage conditions, crypts isolated from colon adenomas derived from Apcmin/+ mice typically grow as spheroidal cysts and do not produce crypts. The adenoma organoid growth requires EGF, but not Wnt, R-spondin or Noggin. However, when mouse colon adenoma spheroids are grown for more than 10 days in the presence of EGF, crypt formation occurs. EGF, EREG, β-cellulin, Neuregulin-1 or AREG are sufficient for initiating crypt formation, however, neuregulin-1 is more potent than the other EGF-family members. EGFR and ErbB2 inhibitors both prevent crypt formation in adenoma cultures. Either EGFR:ErbB2 or ErbB3:ErbB2 signalling is sufficient to initiate adenoma crypt budding and elongation. ErbB2 inhibitors may provide a therapeutic avenue for controlling and ablating colon adenomas.