Novel Venetin-1 nanoparticle from earthworm coelomic fluid as a promising agent for the treatment of non-small cell lung cancer

Abstract

The present research shows the antitumor activity of a protein-polysaccharide complex Venetin-1 obtained from the coelomic fluid of Dendrobaena veneta earthworms against A549 cancer cells. The investigations are a continuation of experiments on the antitumor activity of coelomic fluid obtained from this species. The Venetin-1 nanoparticle was obtained after thermal treatment of the coelomic fluid, separation from coelomocytes, filtration, and lyophilization. The preparation showed a selective effect on cancer cells, whereas normal cells were unaffected. Venetin-1 was effective against the lung cancer cells at doses of 31.3 and 62.5 µg/ml, and the results were imaged using light microscopy and scanning electron microscopy (SEM). The cells died mainly via the apoptosis pathway. Necrotic cells appeared sporadically in the microscopic view. SEM imaging revealed complete destruction of the A549 cells after the incubation with Venetin-1. The atomic force microscopy (AFM) analyses showed changes in the topography, peak force error images, and Young’s modulus (elasticity) of the A549 cells after the incubation with Venetin-1. The transmission electron cryomicroscopy (Cryo-TEM) analysis indicated a polymeric nature of the analyzed preparation. The samples of Venetin-1 showed a very homogeneous size profile with the microparticle size of approximately 58.23 nm. A significant decrease in Venetin-1 binding to sphingomyelin was observed. Venetin-1 lost its pore-forming activity or deactivation of the pore-forming activity occurred. This confirms the absence of hemolytic capacity of Venetin-1 towards red blood cells. The conducted analyses show the suitability of the obtained complex for biomedical research. The next step will consist in analyses of the effect of Venetin-1 on the immune system in mice.

Autophagy of Candida albicans cells after the action of earthworm Venetin-1 nanoparticle with protease inhibitor activity

“…The analysis of the Venetin-1 complex by Cryo-TEM confirmed the nanoparticle size determined by the DLS analysis 25 . The nanoparticles were visible under the microscope as spherical forms, which sometimes coalesced to form double forms indicated by red arrows in Fig.…”

“…Following the analysis of Venetin-1 size (58.23 ± 3.93 nm) 25 conducted with the use of Prometheus Panta, a temperature melting experiment with a temperature gradient from 25 to 95 °C and a heating ramp of 1 °C/min was performed. As shown in Fig.…”

“…such enzymes as proteases 9 – 11 , lysozymes 12 , 13 , metalloenzymes 14 , and fibrinolytic enzymes 15 as well as polysaccharides 16 , proteins 17 – 19 , and nutrients, etc. These factors have antibacterial 20 , 21 , antifungal 8 , 22 , anti-inflammatory 23 , antioxidant 23 , 24 , and anticancer 10 , 11 , 25 – 27 effects.…”

“…Its properties and chemical character were described in earlier publications 49 – 52 . The nanoparticle has been shown to have anticancer 25 , 53 , antiaggregating 54 , and immunostimulating properties 55 . Importantly, it does not show endotoxicity and cytotoxicity to normal human cells 49 .…”

A systematic review of antiproliferative and antitumour activity of earthworm extracts

“…It was stated that the cytotoxic effect of CF of E. eugeniae was superior, followed by CF of P. excavatus. Rybicka et al (2022) investigated the antitumour effects of Venetin-1, a protein-polysaccharide complex obtained from CF of D. veneta, against to lung cancer cell line A549 cells and found that the cells died by apoptosis. In another study with CF of D. veneta, the active protein-carbohydrate fraction isolated from CF disrupted the cell cycle in human colon adenocarcinoma (HT-29 and LS180) cell lines, inhibited 20S proteasome activity, and procaspase-3 (Czerwonka et al 2020).…”