Curcumin-Injected Musca domestica Larval Hemolymph: Cecropin Upregulation and Potential Anticancer Effect.

Shaymaa Mahmoud,Asmaa Hawash,Sami Ullah,Sobhy Hassab El-Nabi,Hesham R El-Seedi,Islam M El-Garawani,Shaden A M Khalifa,Abdullah G Al-Sehemi

doi:10.3390/molecules27051570

Abstract

Over recent decades, much attention has been given to imply the natural products in cancer therapy alone or in combination with other established procedures. Insects have a rich history in traditional medicine across the globe, which holds promise for the future of natural product drug discovery. Cecropins, peptides produced by insects, are components of a defense system against infections and are well known to exert antimicrobial and antitumor capabilities. The present study aimed to investigate, for the first time, the role of curcumin in enhancing the anticancer effect of Musca domestica larval hemolymph. Third larval instars of M. domestica were injected with curcumin and the hemolymph was picked at 4, 8, and 24 h post-curcumin injection. M. domestica cecropin A (MdCecA) was evaluated in control and injected larval hemolymphs. The cytotoxicity on breast cancer cell lines (MCF-7) and normal Vero cells was assessed to be comparable to control larval hemolymph. Curcumin-injected larval hemolymphs exhibited significant cytotoxicity with respect to the uninjected ones against MCF-7; however, Vero cells showed no cytotoxicity. The IC50 was 106 ± 2.9 and 388 ± 9.2 μg/mL for the hemolymphs of injected larvae at 4 and 8 h, respectively, while the control larval hemolymph revealed the IC50 of >500 μg/mL. For mechanistic anticancer evaluation, concentrations of 30, 60, and 100 μg/mL of curcumin-injected larval hemolymphs were examined. A significant G2/M cell cycle arrest was observed, confirming the anti-proliferative properties of hemolymphs over the tested concentrations. The MdCecA transcripts were significantly (p < 0.05) upregulated at 4 and 8 h post-injection, while a significant downregulation was observed after 24 h. Cecropin quantification by LC–MS revealed that MdCecA peptides have the highest expression in the hemolymph of the treated larvae at 8 h relative to the control group. The upregulation of cecropin expression at mRNA and protein levels may be attributed to the curcumin stimulation and linked to the increased cytotoxicity toward the cancer cell line. In conclusion, the results suggest that the apoptotic and anti-proliferative effects of M. domestica hemolymph on MCF-7 cells following the curcumin injection can be used as a natural candidate in future pharmaceutical industries.

Highlights

Natural products have provided successful alternatives in cancer chemotherapy through regular identification of unique potential targets, with specificity or selectivity for cancer cells [1,2]
M. domestica larvae were provided by the Institute of Medical Entomology, Dokki, Giza, Egypt
Changes in M. domestica cecropin A (MdCecA) peptide levels were detected in hemolymphs of control and curcuNmoinm-inojercptehdolloargviacealataldteifrfearteiontnisnwterevraelsobussienrgveNdanaomLoCn/gMtSh(eFgigruoruep1sA. )T. hTheemreo-rtality was rseucltosrrdeevdeawleidthaosuigtnsiifgicnainfitc(aPnt< d0i.0ff5e)riennccreeassbeeitnwteheenpgrortoeuinplsev(0e.l1s i±n 0th.0e4l%ar)v.al hemolymph over all tested time points with respect to the control

Summary

Introduction

Natural products have provided successful alternatives in cancer chemotherapy through regular identification of unique potential targets, with specificity or selectivity for cancer cells [1,2]. M. domestica has exhibited numerous bioactive properties such as antibacterial [9], anti-atherosclerosis and pro-inflammatory responses [10], and immunomodulatory [11], antiviral [12], antifungal [13], antioxidants, and anticancer activities [14]. Numerous cecropins have been identified from different species of insect orders (Hymenoptera, Diptera, Coleoptera, Lepidoptera, and Isoptera) [18,19] and other organisms including mammals [20]. These are α-helix linear peptides of approximately 30–35 amino acid residues without cysteine residues and are divided as cecropins A, B, and D [21,22]. They are relatively small proteins that are active against both Gram-positive and Gram-negative bacteria. Cecropin with anti-inflammatory activity has been identified in butterfly, Papilio xuthus [33], and Black fly, Simulium bannaense, salivary glands [34]

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Conclusion