Antitumor Activity of Nitazoxanide against Colon Cancers: Molecular Docking and Experimental Studies Based on Wnt/β-Catenin Signaling Inhibition

Abstract

In colon cancer, wingless (Wnt)/β-catenin signaling is frequently upregulated; however, the creation of a molecular therapeutic agent targeting this pathway is still under investigation. This research aimed to study how nitazoxanide can affect Wnt/β-catenin signaling in colon cancer cells (HCT-116) and a mouse colon cancer model. Our study included 2 experiments; the first was to test the cytotoxic activity of nitazoxanide in an in vitro study on a colon cancer cell line (HCT-116) versus normal colon cells (FHC) and to highlight the proapoptotic effect by MTT assay, flow cytometry and real-time polymerase chain reaction (RT-PCR). The second experiment tested the in vivo cytotoxic effect of nitazoxanide against 1,2-dimethylhydrazine (DMH) prompted cancer in mice. Mice were grouped as saline, DMH control and DMH + nitazoxanide [100 or 200 mg per kg]. Colon levels of Wnt and β-catenin proteins were assessed by Western blotting while proliferation was measured via immunostaining for proliferating cell nuclear antigen (PCNA). Treating HCT-116 cells with nitazoxanide (inhibitory concentration 50 (IC50) = 11.07 µM) revealed that it has a more cytotoxic effect when compared to 5-flurouracil (IC50 = 11.36 µM). Moreover, it showed relatively high IC50 value (non-cytotoxic) against the normal colon cells. Nitazoxanide induced apoptosis by 15.86-fold compared to control and arrested the cell cycle. Furthermore, nitazoxanide upregulated proapoptotic proteins (P53 and BAX) and caspases but downregulated BCL-2. Nitazoxanide downregulated Wnt/β-catenin/glycogen synthase kinase-3β (GSK-3β) signaling and PCNA staining in the current mouse model. Hence, our findings highlighted the cytotoxic effect of nitazoxanide and pointed out the effect on Wnt/β-catenin/GSK-3β signaling.

Nitazoxanide inhibits acetylated KLF5-induced bone metastasis by modulating KLF5 function in prostate cancer

“…Although our study is the first to demonstrate a therapeutic effect of NTZ on the bone metastasis of prostate cancer, its anticancer activity has been reported in recent studies in different types of cancers involving various molecular mechanisms. For example, it suppressed colon tumor growth likely by inducing cell cycle arrest and altering the expression of multiple molecules [52,53]; it suppressed ovarian cancer growth partially by inhibiting the protein disulfide isomerase (PDI) activity [54]; it inhibited sphere formation in 3D cultures of HCC and CRC cells involving the inhibition of OXPHOS [55,56]; and it suppressed glioma growth by causing the G2/M cell cycle arrest, inducing apoptosis, and inhibiting autophagy likely via CDK1 inhibition, ING1 upregulation, etc. [57,58].…”

Formulation and Characterization of Doxycycline-Loaded Polymeric Nanoparticles for Testing Antitumor/Antiangiogenic Action in Experimental Colon Cancer in Mice

“…Histologic scoring was used to determine the degree of dysplasia using previously stabilized parameters. In summary, tissue specimens were classified as having no dysplastic changes or having dysplastic changes: non-dysplasia—goblet cells that appear normal (basal-oriented nuclei and apical localization of mucus); mild dysplasia—hypercellularity of elongated cells and localized nuclear stratification were observed (hyperchromasia) [ 48 ].…”

Salicylanilides and Their Anticancer Properties

“…Moreover, Qu et al reported that nitazoxanide also could suppress Wnt/β-catenin signaling through a specific target, PAD2, which is an enzyme responsible for protein citrullination, and thus induces citrullination of β-catenin [ 46 ]. Recently, a number of studies further reported that nitazoxanide downregulated the Wnt/β-catenin signaling pathway [ 48 , 130 , 131 ].…”