The Anti-Tumoral Potential of Phosphonate Analog of Sulforaphane in Zebrafish Xenograft Model

Abstract

Isothiocyanates (ITCs) show strong activity against numerous human tumors. Five structurally diverse ITCs were tested in vivo using the zebrafish embryos 6 and 48 h post-fertilization (hpf). The survival rate, hatching time, and gross morphological changes were assessed 24, 48, and 72 h after treatment with all compounds in various doses (1–10 µM). As a result, we selected a phosphonate analog of sulforaphane (P-ITC; 1–3 µM) as a non-toxic treatment for zebrafish embryos, both 6 and 48 hpf. Furthermore, the in vivo anti-cancerogenic studies with selected 3 µM P-ITC were performed using a set of cell lines derived from the brain (U87), cervical (HeLa), and breast (MDA-MB-231) tumors. For the experiment, cells were labeled using red fluorescence dye Dil (1,1′-Dioctadecyl-3,3,3′,3′-Tetramethylindocarbocyanine, 10 μg/mL) and injected into the hindbrain ventricle, yolk sac region and Cuvier duct of zebrafish embryos. The tumor size measurement after 48 h of treatment demonstrated the significant inhibition of cancer cell growth in all tested cases by P-ITC compared to the non-treated controls. Our studies provided evidence for P-ITC anti-cancerogenic properties with versatile activity against different cancer types. Additionally, P-ITC demonstrated the safety of use in the living organism at various stages of embryogenesis.

Isothiocyanates (ITCs) 1-(Isothiocyanatomethyl)-4-phenylbenzene and 1-Isothiocyanato-3,5-bis(trifluoromethyl)benzene—Aldehyde Dehydrogenase (ALDH) Inhibitors, Decreases Cisplatin Tolerance and Migratory Ability of NSCLC

“…18 , 19 , and 36 : these were found to manifest strong affinity to ALDH isoforms and ABC proteins based on in silico analysis. Interestingly, 18 and 19 have also presented strong anti-tumoral properties in a xenograft zebrafish model [ 83 ]. The chosen ITCs appear to be strong anti-cisplatin-resistance agents: they were found to significantly repress cisplatin tolerance in both A549CisR and in the stage 4 NSCLC cell line NCI-H1581 at concentrations 10-fold lower than DSF.…”

“…ITCs have also been found to be effective against a murine mammary gland carcinoma 4T1 model in vivo, with administration resulting in reduced tumor mass [ 33 ]. Furthermore, 3 µM ITC analogs inhibited the motility of three highly malignant cell lines derived from cervical (HeLa), glioblastoma (U87), and breast (MDA-MB-231) carcinomas [ 83 ]. This finding is consistent with our present observations.…”

Sulforaphane and Its Bifunctional Analogs: Synthesis and Biological Activity

“…In vivo studies of the selected P-ITCs showed slightly lower anticancer and antimetastatic activity in comparison to naturally occurring BITC. However, recent studies in vivo on Zebrafish have shown that P-SFN 291 was characterized by high anticancer activity and low toxicity [ 256 ].…”

Development of a Triple-Negative Breast Cancer Leptomeningeal Disease Model in Zebrafish

“…The optical clarity of the zebrafish ( Danio rerio ) model makes it uniquely suited for the study of TNLMD as it enables microinjection of cancer cells into LMD-associated anatomical structures while allowing simultaneous fluorescent monitoring of cell proliferation and migration [ 12 , 13 ]. A major advantage of zebrafish xenograft models over rodent models is the availability of pigment-deficient strains, e.g., casper ( roy -/-; nacre -/-), that allow observation of transplanted cancer cells, and the absence until 4–6 weeks after fertilization of an adaptive immune system that could reject foreign implanted cells [ 11 , 14 , 15 ].…”

“…Microinjection of fluorescent rhodamine dye into the CSF of the hindbrain ventricle permits microscopic visualization and analysis of the developing zebrafish ventricular system [ 16 , 17 , 18 ]. Researchers have microinjected glioblastoma cells labeled with the red fluorescent dye 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine (DiI) into the zebrafish hindbrain ventricle and assessed cancer cell proliferation and migration using bright-field and fluorescent microscopy, and analyzed glioblastoma response to the phosphoinositide 3-kinase inhibitor LY294002, zinc oxide nanoparticles, and isothiocyanate derivatives [ 12 , 13 ]. In addition, transplantation of zebrafish brain tumor cells into the fourth ventricle resulted in LMD along the spinal cord in a subset of recipients [ 19 ].…”