Zebrafish Stem Cell Differentiation Stage Factors Suppress Bcl-xL Release and Enhance 5-Fu-Mediated Apoptosis in Colon Cancer Cells

F.      D'Anselmi; A.      Cucina; P.      M. Biava; S.      Proietti; P.      Coluccia; L.      Frati; M.      Bizzarri

doi:10.2174/138920111794295864

Abstract

Stem cell differentiation stage factors (SCDSF), taken from Zebrafish embryos during the stage in which totipotent stem cells are differentiating into pluripotent stem cells, have been shown to inhibit proliferation and induce apoptosis in colon tumors. In order to ascertain if these embryonic factors could synergistically/additively interact with 5- Fluorouracil (5-Fu), whole cell-count, flow-cytometry analysis and apoptotic parameters were recorded in human colon cancer cells (Caco2) treated with Zebrafish stem cell differentiation stage factors (SCDSF 3 μg/ml) in association or not with 5-Fu in the sub-pharmacological therapeutic range (0.01 mg/ml). Cell proliferation was significantly reduced by SCDSF, meanwhile SCDSF+5-Fu leads to an almost complete growth-inhibition. SCDSF produces a significant apoptotic effect, meanwhile the association with 5-FU leads to an enhanced additive apoptotic rate at both 24 and 72 hrs. SCDSF alone and in association with 5-Fu trigger both the extrinsic and the intrinsic apoptotic pathways, activating caspase-8, -3 and -7. SCDSF and 5-Fu alone exerted opposite effects on Bax and Bcl-xL proteins, meanwhile SCDSF+5-Fu induced an almost complete suppression of Bcl-xL release and a dramatic increase in the Bax/Bcl-xL ratio. These data suggest that zebrafish embryo factors could improve chemotherapy efficacy by reducing anti-apoptotic proteins involved in drug resistance processes.

Keywords: Apoptosis, Bcl-xL, colon cancer, 5-Fluorouracil, zebrafish stem cell differentiation stage factors, Stem cell differentiation stage factors (SCDSF), pluripotent stem cells, totipotent stem cells, proliferation, flow-cytometry analysis, chemotherapy efficacy, anti-apoptotic proteins, drug-resistance processes, embryonic tissues, cytostatic drugs, phenotype, chemosensitivity, Fetal Calf Serum, Streptomycin, Penicillin, Gentamycin, sulforhodamine B, ELISA, histone-associated DNA fragments, anti-alpha-tubulin, Optical density (O.D.), Srb-c assay, cytofluorimetric assays, density (O.D.), caspase-3 activation, Bax protein, embryonic mi-cro-environment, morphogenesis, angiogenesis, anti-sense oligonucleotides, curcumin