Cytotoxicity against tumor cell lines and anti-inflammatory properties of chitinases from Calotropis procera latex

Abstract

The role of chitinases from the latex of medicinal shrub Calotropis procera on viability of tumor cell lines and inflammation was investigated. Soluble latex proteins were fractionated in a CM Sepharose Fast-Flow Column and the major peak (LPp1) subjected to ion exchange chromatography using a Mono-Q column coupled to an FPLC system. In a first series of experiments, immortalized macrophages were cultured with LPp1 for 24 h. Then, cytotoxicity of chitinase isoforms (LPp1-P1 to P6) was evaluated against HCT-116 (colon carcinoma), OVCAR-8 (ovarian carcinoma), and SF-295 (glioblastoma) tumor cell lines in 96-well plates. Cytotoxic chitinases had its anti-inflammatory potential assessed through the mouse peritonitis model. We have shown that LPp1 was not toxic to macrophages at dosages lower than 125 μg/mL but induced high messenger RNA expression of IL-6, IL1-β, TNF-α, and iNOs. On the other hand, chitinase isoform LPp1-P4 retained all LPp1 cytotoxic activities against the tumor cell lines with IC ranging from 1.2 to 2.9 μg/mL. The intravenous administration of LPp1-P4 to mouse impaired neutrophil infiltration into the peritoneal cavity induced by carrageenan. Although the contents of pro-inflammatory cytokines IL-6, TNF-α, and IL1-β were high in the bloodstreams, such effect was reverted by administration of iNOs inhibitors NG-nitro-L-arginine methyl ester and aminoguanidine. We conclude that chitinase isoform LPp1-P4 was highly cytotoxic to tumor cell lines and capable to reduce inflammation by an iNOs-derived NO mechanism.

Anticancer and antifungal efficiencies of purified chitinase produced from <i>Trichoderma viride</i> under submerged fermentation

“…Similar results are recorded by Viana et al (2017) who reported that cytotoxicity of chitinase isoforms from Calotropis procera latex against HCT-116, OVCAR-8 and SF-295 (glioblastoma) tumor cell lines with IC50 ranging from 1.2~2.9 mg/ml. The real mechanism of the anticancer effect of chitinase is not clear yet, thus future studies are necessary.…”

Latex proteins downregulate inflammation and restores blood-coagulation homeostasis in acute Salmonella infection

“…(23) This cytokine will activate inflammatory tissue and recruit macrophages and neutrophils, similar to observed with LP PI . (3) Salmonella causes the release of chemokines and cytokines that provoke a local inflammatory response, attracting neutrophils and macrophages to use as a new host to disseminate itself through the body. Whereas Salmonella survives inside macrophages, neutrophils are able to kill the bacteria, due to the high concentra- tion of ROS in the phagosome of these cells.…”

“…These effects include analgesic, anti-inflammatory, proinflammatory, antimicrobial activity, and others. (3,4) Among the constituents of C. procera latex, approximately 85% of its dry mass is rubber (polyisoprene), while soluble proteins represent around 10%. Among the soluble laticifer proteins (LP), are peptidases, that degrade substate BANA, and azocasein in neutral pH.…”

An Overview of the Characteristics and Potential of Calotropis procera From Botanical, Ecological, and Economic Perspectives

“…Calotropis procera causes acute toxicity in various plant and animal cells, including human beings. Different plant parts, particularly the latex, are therefore tested against various cancer cell lines (Ibrahim et al, 2015;Viana et al, 2017;Al-Qahtani et al, 2020). Similarly, antibacterial and antihelminitic potential of the plant is being utilized in pharmacology (details provided in section "Pharmacological Applications").…”