Determination of anti-cancer and antioxidant properties of protein extracts obtained from aquatic Helophorus (Coleoptera: Helophoridae) insects

Tuba Elhazar; Bülent Kaya; Fatma Caf

doi:10.12714/egejfas.40.1.05

TR EN

Sucul Helophorus (Coleoptera: Helophoridae) böceklerinden elde edilen protein ekstraktlarının anti-kanser ve antioksidan özelliklerinin belirlenmesi

Öz

Bu çalışmada, Helophorus (Coleoptera: Helophoridae) cinsine ait sucul böcek Helophorus aquaticus ve Helophorus syriacus türlerinden protein ekstraksiyonu yapılmıştır. Böcek türleri 2017 Mayıs-Haziran ayları arasında ve Bingöl il merkezi ve ilçelerinden çeşitli akarsu, kaynak, dere, birikinti ve sıcak su gözelerinin sığ kesimlerinden toplandı. Protein miktarı H. aquaticus’da 34,78 mg/mL, H. syriacus’da 35,14 mg/mL olarak belirlenmiştir. Daha sonra elde edilen protein ekstraktlarının antioksidan kapasitesi incelendi. Metal şelatlama aktivitesi H. aquaticus ve H. syriacus için sırasıyla %90-88,5, DPPH uzaklaştırma aktivitesi %53,19-61,7 olarak belirlendi. Her iki türe ait protein numuneleri, in vitro hücre kültürü ile PC-3 (prostat kanseri) hücrelerinde WST-1 ile hücre canlılığı açısından test edildi. Test sonuçlarının incelenmesi üzerine, çalışılan türlerin her ikisinden alınan protein ekstraktları, hücre inhibisyonunda bir azalmaya neden oldu. En yüksek hücre inhibisyonu, 1000 μg/mL böcek proteini ekstresi eklenen numunelerde gözlendi. Bu çalışmada, etkin doz belirlendikten sonra Western blot tekniği ile apoptozu sağlayan protein ekspresyonu incelenmiştir. Western blot tekniği ile Cyt-C ve Caspase-3 proteinlerine bakılarak, apoptoz dışı hücre ölümünde in vitro PC-3 hattı için her iki türden protein ekstraktlarının etkinliği etkili bir şekilde gösterildi. Çalışma sonucunda böcek proteinlerinin western blot tekniği ile hücre ölümünü sağlayan proteinlerin üretimini desteklediği gösterildi.

Anahtar Kelimeler

Determination of anti-cancer and antioxidant properties of protein extracts obtained from aquatic Helophorus (Coleoptera: Helophoridae) insects

Abstract

In this study, protein extraction was performed from the aquatic insect Helophorus aquaticus (Linnaeus, 1758) and Helophorus syriacus (Kuwert, 1885) species belonging to the genus Helophorus (Coleoptera: Helophoridae). Then, these protein extracts were studied in vitro. These species were collected from the shallow parts of various streams, springs, creeks, ponds and hot springs from Bingöl city centre and its districts between May to June 2017. The protein amount was determined 34.78 mg/mL in the H. aquaticus and 35.14 mg/mL in the H. syriacus, after that he antioxidant capacity of protein extracts was examined. Metal chelating activity was determined as 90-88.5% and DPPH removal activity 53.19-61.7% for H. aquaticus and H. syriacus respectively. Protein samples belonging to both species were tested for cell vitality with WST-1 in PC-3 (prostate cancer) cells with in vitro cell culture. Upon examination of the test results, it has been found out that protein extracts from both of the studied species caused a decrease in cell inhibition. The highest cell inhibition was observed in samples with 1000 μg/mL insect protein extract added. In this study, protein expression providing apoptosis was examined with the Western blot technique after the effective dose was established. By looking at the proteins of Cyt-C and Caspase 3 with the Western blot technique, the efficacy of the protein extracts from both species was demonstrated effectively for the in vitro PC-3 line in non-apoptosis cell death. As a result of the study, insect proteins were shown to support the production of proteins that ensure cell death with the western blot technique.

Keywords

Supporting Institution

This works were supported by the Science Research Foundation of Bingol University of Turkey

Project Number

Project number: BAP-FEF.2017.00.003

Thanks

We thank Abdullah Mart for the identification of the species. We would also like to thank Ekrem Darendelioğlu for his help and laboratory for the Western Blot part of the study.

References

Berenbaum, M.R. (1993). Sequestered plant toxins and insect palatability. The Food Insects Newsletter, 6(3), 6-9.
Blum, M.S., 1994. The limits of entomophagy: a discretionary gourmand in a world of toxic Insects. The Food Insects Newsletter, 7(1), 6-11.
Bulet, P., Stöcklin, R., & Menin. L. (2004). Anti-microbial peptides: from invertebrates to vertebrates. Immunological Reviews, 198, 169-184. https://doi.org/10.1111/j.0105-2896.2004.0124.x
Caf, F., Yildiz, G., Özdemir, N.S., & Mart A. (2020). A chemotaxonomic approach to fatty acid composition of the genera Helochares Mulsant, 1844 and Coelostoma Brullé, 1835 (Coleoptera: Hydrophilidae). Turkish Journal of Entomology, 44(3), 399-412. https://doi.org/10.16970/entoted.657190
Chernysh, S., Kim, S.I., Bekker, G., Pleskach, V.A., Filatova, N.A., Anikin, V.B., & Bulet, P. (2002). Antiviral and antitumor peptides from insects. Proceedings of the National Academy of Sciences, 99(20), 12628-12632. https://doi.org/10.1073/pnas.192301899
Charbonnier, S., Gallego, O., & Gavin, A.C. (2008). The social network of a cell: Recent advances in interactome mapping. Biotechnology Annual Review, 14, 1-28. https://doi.org/10.1016/S1387-2656(08)00001-X
Colell, A., Ricci, J.E., Tait S., Milasta, S., Maurer, U., Bouchier-Hayes, L., Fitzgerald, P., Guio-Carrion, A., Waterhouse, N.J., Li, C.W., Mari, B., Barbry, P., Newmeyer, D.D., Beere, H.M., & Green D.R. (2007). GAPDH and autophagy preserve survival after apoptotic cytochrome c release in the absence of caspase activation. Cell, 129(5), 983-997. https://doi.org/10.1016/j.cell.2007.03.045
Costa-Neto, E.M. (2002). The use of insects folk medicine in the State of Bahai, Northeastern Brazil, with notes on insects reported elsewhere in Brazilian folk medicine. Human Ecology, 30(2), 254-263. https://doi.org/10.1023/A:1015696830997

Coşkun, M., Kayis, T., Ozalp, P., Kocalar, K., Tatlicioğlu, C. I., & Emre, I. (2009). The effects of a meridic diet on the sex ratio of offspring, on glycogen and protein content, and on productivity and longevity of adult Pimpla turionellae (Hymenoptera: Ichneumonidae) for five generations. Belgian Journal of Zoology, 139(2), 103-108.
Darılmaz, M. (2010). Investigation of Inner West Anatolia Aquatic Coleoptera Fauna. Doctoral dissertation, Gazi University Institute of Science and Technology, Turkey (in Turkish)
da Silva Lucas, A. J., de Oliveira, L. M., da Rocha, M., & Prentice, C. (2020). Edible insects: an alternative of nutritional, functional and bioactive compounds. Food Chemistry, 311, 126022. https://doi.org/10.1016/j.foodchem.2019.126022
Defoliart, G.R. (1995). Edible insects as minilivestock. Biodiversity and Conservation, 4, 306-321. https://doi.org/10.1007/BF00055976
Di Mattia, C., Battista, N., Sacchetti, G., & Serafini, M. (2019). Antioxidant activities in vitro of water and liposoluble extracts obtained by different species of edible insects and invertebrates. Frontiers in Nutrition, 6, (106). https://doi.org/10.3389/fnut.2019.00106
Doucet, C.M., & Hetzer, M.W. (2010). Nuclear pore biogenesis into an intact nuclear envelope. Choromosoma, 119, 469-477. https://doi.org/10.1007/s00412-010-0289-2
Duffey, S.S. (1980). Sequestration of plant natural products by insects. Annual Review of Entomology, 25, 447-477. https://doi.org/10.1146/annurev.en.25.010180.002311
Fikáček M., Schmied H., & Prokop, J. (2010). Fossil hydrophilid beetles (Coleoptera: Hydrophilidae) of the Late Oligocene Rott Formation (Germany). Acta Geologica Sinica 84 (4), 732-750. https://doi.org/10.1111/j.1755-6724.2010.00239.x
Gil, M.I., Tomás-Barberán, F.A., Hess-Pierce, B., Holcroft, D.M., & Kader, A.A. (2000). Antioxidant activity of pomegranate juice and its relationship with phenolic composition and processing. Journal of Agricultural and Food Chemistry 48, 4581-4589. https://doi.org/10.1021/jf000404a
Hatano, T., Kagawa, H., Yasuhara, T., & Okuda, T. (1988). Two new flavonoids and other constituents in licorice root: their relative astringency and radical scavenging effects, Chemical and Pharmaceutical Bulletin, 36(6), 2090-2097. https://doi.org/10.1248/cpb.36.2090
Jantzen da Silva Lucas, A., Menegon de Oliveira, L., da Rocha, M., & Prentice, C. (2020). Edible insects: An alternative of nutritional, functional and bioactive compounds. Food Chemistry, 311, 126022. https://doi.org/10.1016/j.foodchem.2019.126022
Karaś, M., Baraniak, B., Rybczynska, K., Gminski, J., Gaweł- Bezben, K., & Jakubczyk, A. (2015). The influence of heat treatment of chickpea seeds on antioxidant and fibroblast growth-stimulating activity of peptide fractions obtained from proteins digested under simulated gastrointestinal conditions. International Journal of Food Science & Technology, 50, 2097-2103. https://doi.org/10.1111/ijfs.12872
Koczulla, A.R., & Bals, R. (2003). Antimicrobial peptides-current status and therapeutic potentials. Drugs, 63, 389-406. https://doi.org/10.2165/00003495-200363040-00005
Koç, K. (2011). Investigation of In vitro Genotoxic and Oxidative Effects of Some Edible Insects, Master Thesis, Atatürk University, Institute of Science, Erzurum (in Turkish)
Koç, K., İncekara, Ü., Türkez, H., & Çelik, K. (2019). In vitro Assessment of Genotoxic And Oxidative Effects Potentials Of Edible Bamboo Worms And Weaver Ants. Munis Entomology and Zoology Journal, 14(2), 496-501.
Liu S., Sun J., Yu L., Zhang C., Bi J., Zhu F., Qu M., & Yang Q. (2012). Antioxidant activity and phenolic compounds of Holotrichia parallela Motschulsky extracts. Food Chemistry 134, 1885-1891. https://doi.org/10.1016/j.foodchem.2012.03.091
Mart, A. (2009). Water scavenger beetles (Coleoptera: Hydrophilidae) provinces of Central Black Sea region of Turkey. Journal of The Entomological Reserch Society, 11(1), 47-70.
Mittapalli, O., Neal, J.J., & Shukle, R.H. (2007). Antioxidant defense response in a galling insect. Proceedings of the National Academy of Sciences, 104(6), 1889-1894. https://doi.org/10.1073/pnas.0604722104
McKenzie, E.A., & Abbott, W.M. (2018). Expression of recombinant proteins in insect and mammalian cells. Methods, 147, 40-49. https://doi.org/10.1016/j.ymeth.2018.05.013
Polat, A., Darılmaz, M.C. & İncekara, Ü. (2021). An annotated checklist of the Hydrophiloidea (Coleoptera) of Turkey. Munis Entomology & Zoology, 16 (1), 151-178.
Porter, A.G., & Jänicke, R.U. (1999). Emerging roles of caspase-3 in apoptosis. Cell Death and Differentiation, 6, 99-145. https://doi.org/10.1038/sj.cdd.4400476
Riccio, A. (2010). Dynamic epigenetic regulation in neurons: enzymes, stimuli and signaling pathways. Nature neuroscience, 13, 1330-1337. https://doi.org/10.1038/nn.2671
Stuible, M., Burlacu, A., Perret, S., Brochu, D., Paul-Roc, B., Baardsnes, J., Loignon, M., Grazzini, E., & Durocher, Y. (2018). Optimization of a high-cell-density polyethylenimine trasfection method for rapid protein production in CHO-EBNA1 cells. Journal of Biotechnology, 281, 39-47. https://doi.org/10.1016/j.jbiotec.2018.06.307
Suh, H. J., Kim, S. R., Lee, K. S., Park, S., & Kang, S.C. (2010). Antioxidant activity of various solvent extracts from Allomyrina dichotoma (Arthropoda: Insecta) larvae. Journal of Photochemistry and Photobiology B: Biology, 99(2), 67-73. https://doi.org/10.1016/j.jphotobiol.2010.02.005
Torres-Fuentes, C., Alaiz, M., & Vioque, J. (2011). Affinity purification and characterisation of chelating peptides from chickpea protein hydrolysates. Food Chemistry, 129, 485-490. https://doi.org/10.1016/j.foodchem.2011.04.103
Yılmaz, A. (2011). Faunistic and Systematic Investigation of Helophoridae, Hydrophilidae (Coleoptera) Species in Ista Province, Master Thesis, Süleyman Demirel University, Institute of Science, Isparta (in Turkish)
Van Huis, A. (2013). Potential of insects as food and feed in assuring food security. Annual review of entomology, 58, 563-583. https://doi.org/10.1146/annurev-ento-120811-153704
Wenli, Y., Yaping, Z., & Bo, S, (2004). The radical scavenging activities of radix puerariae isoflavonoids: A chemiluminescence study. Food Chemistry, 86(4), 525-529. https://doi.org/10.1016/j.foodchem.2003.09.005
Wu, Q.Y., Jia, J.Q., Tan, G.X., Xu, J.L., & Gui, Z.Z. (2011). Physicochemical properties of silkworm larvae protein isolate and gastrointestinal hydrolysate bioactivities. African Journal of Biotechnology, 10, 6145-6153.
Zieliǹska, E., Baraniak, B., Karas, M., Rybczynska, K., & Jakubczyk, A. (2015). Selected species of edible insects as a source of nutrient composition. Food Research International, 77, 460-466. https://doi.org/10.1016/j.foodres.2015.09.008
Zielińska, E., Karaś, M., & Jakubczyk, A. (2017). Antioxidant activity of predigested protein obtained from a range of farmed edible insects. International Journal of Food Science & Technology, 52, 306-312. https://doi.org/10.1111/ijfs.13282
Zielińska, E., Karaś, M., & Baraniak, B. (2018). Comparison of functional properties of edible insects and protein preparations thereof. LWT - Food Science and Technology, 91, 168-174. https://doi.org/10.1016/j.lwt.2018.01.058

Details

Primary Language

English

Subjects

Structural Biology

Journal Section

Research Article

Authors

Tuba Elhazar
0000-0003-3632-1159
Türkiye

Bülent Kaya
0000-0002-1216-6441
Türkiye

Fatma Caf ^*
0000-0002-0363-4848
Türkiye

Publication Date

March 15, 2023

Submission Date

September 2, 2022

Acceptance Date

January 13, 2023

Published in Issue

Year 2023 Volume: 40 Number: 1

DOI

https://doi.org/10.12714/egejfas.40.1.05

IZ

https://izlik.org/JA75UL93AR

Cite

RIS / Bibtex

APA

Elhazar, T., Kaya, B., & Caf, F. (2023). Determination of anti-cancer and antioxidant properties of protein extracts obtained from aquatic Helophorus (Coleoptera: Helophoridae) insects. Ege Journal of Fisheries and Aquatic Sciences, 40(1), 35-42. https://doi.org/10.12714/egejfas.40.1.05

Cited By

Revisiting edible insects as sources of therapeutics and drug delivery systems for cancer therapy

Frontiers in Pharmacology

https://doi.org/10.3389/fphar.2024.1345281