Mesobuthus gibbosus (Scorpiones: Buthidae) Akrep Ham Zehrinin Kanser Hücre Hatları Üzerine Sitotoksik ve Antiproliferatif Etkilerinin Araştırılması

Amaç: Akrep zehirleri, çeşitli kimyasal maddelerin oluşturduğu oldukça kompleks heterojen karışımlardır. Bu çalışmada, Mesobuthus gibbosus (Brullé, 1832) (Scorpiones: Buthidae) akrebinin ham zehrinin kanser hücreleri üzerine sitotoksik ve antiproliferatif etkileri araştırılmıştır. Gereç ve Yöntemler: M. gibbosus’dan sağılan ham zehrin sitotoksik etkileri MCF-7 meme kanseri ve A549 akciğer karsinomu ve normal fare fibroblast (L929) hücreleri üzerindeki sitotoksik ve antiproliferatif etkileri çalışılmış ve elde edilen veriler değerlendirilmiştir. Sitotoksisiteyi belirlemek için MTT testi, apoptoz ve nekroz indeksini belirlemek için ikili boyama metodu ve xCELLigence gerçek zamanlı analiz sistemi ile hücre proliferasyonunun analizi in vitro koşullarda yapılmıştır. Bulgular: Çalışmada, akrep ham zehrinin MCF-7 ve A549 kanserli hücrelerin üzerine sitotoksik etki sonuçları ayrı ayrı L929 fibroblast hücre sonuçları ile Mann Whitney U testi kullanılarak istatistiksel analize tabi tutulmuştur. Bunun sonucunda L929 hücrelerinin % canlılık değeri A-549 ‘un değerine çok yakın iken MCF-7 hücrelerinin % canlılık değeri L929’un değerine göre daha yüksek olmasına rağmen araştırmadaki fark, istatistiksel olarak anlamlı bulunmamıştır. Bu hücrelerin proliferasyon grafikleri incelendiğinde ise en yüksek dozdaki zehrin MCF-7 ve L929 hücrelerinin proliferasyonunun bir miktar azalttığı ancak A549 hücresinin proliferasyonunu etkilemediği gözlenmiştir. Sonuç: M. gibbosus akrep ham zehrinin MCF-7 ve A549 kanserli hücreleri üzerine apoptotik ve antiproliferatif etkileri bir miktar gözlenmiştir. Ancak bu sonuçlar istatiksel olarak anlamlı değildir.

Anahtar Kelimeler:

Akrep Zehri, Mesobuthus gibbossus, Kanser, Hücre Proliferasyonu, Sitotoksisite

Investigation of Cytotoxic and Antiproliferative Effects of Mesobuthus gibbosus (Scorpiones: Buthidae) Crude Scorpion Venom on Cancer Cell Lines

Objective: Scorpion venoms are a highly complex mixture of various chemicals with surprising biological effects. In this study, the cytotoxic and anti-proliferative effects of Mesobuthus gibbosus (Brullé, 1832) (Scorpiones: Buthidae) scorpion crude venom on cancer cell lines were investigated. Materials and Methods: The cytotoxic and anti-proliferative effects of crude venom milked from M. gibbosus on MCF-7 breast cancer and A549 lung carcinoma, and normal mouse fibroblast (L929) cells were studied and the obtained data were evaluated. The MTT test to determine cytotoxicity, the double staining method to determine the apoptosis and necrosis index, and the analysis of cell proliferation with the xCELLigence real-time analysis system were performed in vitro. Results: In the study, the results of cytotoxic effects of scorpion crude venom on between MCF-7 and A549 cancer cells and L929 fibroblast cell were analyzed statistically by using Mann Whitely U test separately. As a result, although the % viability value of L929 cells was very close to the value of A-549, the % viability value of MCF-7 cells was higher than the value of L929, but the difference in the study was not statistically significant. When the proliferation graphs of these cells were examined, it was observed that the highest dose of venom slightly reduced the proliferation of MCF-7 and L929 cells, but did not affect the proliferation of A549 cells. Conclusion: The apoptotic and antiproliferative effects of M. gibbosus scorpion crude venom on MCF-7 and A549 cancer cells were observed to some extent. However, these results are not statistically significant.

Keywords:

Scorpion Venom, Mesobuthus gibbosus, Cancer, Cell Proliferation, Cytotoxic,

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1. Sağlık Bakanlığı Tedavi Hizmetleri Genel Müdürlüğü. Türkiye Onkoloji Hizmetleri Yeniden Yapılanma Programı 2010-2023. Ankara: T.C. Sağlık Bakanlığı. 2010.
2. Tuncer M. Globalleşen Kanser ve Türkiye. 2008. www.nukte.org.
3. Brawley OW. A brief history of the American cancer society extramural research program. CA Cancer J Clin. 2013; 63(1):9-10.
4. Baskar R, Lee KA, Yeo R. Cancer and radiation therapy:current advances and future directions. International Journal of Medical Sciences. 2012; 9(3): 193-199.
5. Santibáñez-López CE, Possani LD. Overview of the Knottin scorpion toxin-like peptides in scorpion venoms: Insights on their classification and evolution. Toxicon. 2015; 107(Pt B): 317-326.
6. Rein JO. https://www.ntnu.no/ub/scorpion-files/ 2022.
7. Ma Y, He Y, Zhao R, Wu Y, Li W, Cao Z. Extreme diversity of scorpion venom peptides and proteins revealed by transcriptomic analysis: implication for proteome evolution of scorpion venom arsenal. J. Proteomics. 2012; 75(5): 1563-1576.
8. Rodríguez de la Vega RC, Schwartz EF, Possani LD. Mining on scorpion venom biodiversity. Toxicon. 2010; 56(7): 1155-1161.
9. Almaaytah A, Tarazi S, Mhaidat N, Al-Balas Q, Mukattash TL. Mauriporin, a novel cationic α-helical peptide with selective cytotoxic activity against prostate cancer cell lines from the venom of the scorpion Androctonus mauritanicus. Int. J. Pept. Res. Ther. 2013; 19(4): 281-293.
10. Nabi G, Ahmad N, Ullah S, Khan S. Therapeutic Applications of Scorpion Venom in Cancer: Mini Review. Journal of Biology and Life Sciences. 2015; 6(1): 57.
11. Bradford MM. Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry. 1976; 72: 248-254.
12. Ucar G, Tas C, Tümer A. Monoamine oxidase inhibitory activities of the scorpion Mesobuthus gibbosus (Buthidae) venom peptides. Toxicon. 2005; 45(1): 43-52.
13. Ucar G, Tas C. Cholinesterase inhibitory activities of the scorpion Mesobuthus gibbosus (Buthidae) venom peptides. FABAD J Pharm Sci. 2003; 28(1): 61-70.
14. Diego-García, E, Caliskan F, Tytgat J. The Mediterranean scorpion Mesobuthus gibbosus (Scorpiones, Buthidae): transcriptome analysis and organization of the genome encoding chlorotoxin-like peptides. BMC genomics. 2014; 15(1): 1-16.
15. Feng L, Gao R, Gopalakrishnakone P. Isolation and characterization of a hyaluronidase from the venom of Chinese red scorpion Buthus martensi. Comparative Biochemistry and Physiology Part C. Toxicology & Pharmacology. 2008: 148(3), 250-257.
16. Kesavan K, Ratliff J, Johnson EW, Dahlberg W, Asara JM, Misra P, ... & Jacoby DB. Annexin A2 is a molecular target for TM601, a peptide with tumor-targeting and anti-angiogenic effects. Journal of Biological Chemistry. 2010; 285(7): 4366-4374.
17. Veiseh M, Gabikian P, Bahrami SB, Veiseh O, Zhang M, Hackman, RC, ... & Olson JM. Tumor paint: α- chlorotoxin: Cy5. 5 bioconjugate for intraoperative visualization of cancer foci. Cancer research. 2007: 67(14): 6882-6888.
18. Heinen TE, da Veiga ABG. Arthropod venoms and cancer. Toxicon. 2011; 57(4): 497-511.
19. Wang WX, Ji YH. Scorpion venom induces glioma cell apoptosis in vivo and inhibits glioma tumor growth in vitro. Journal of neuro-oncology. 2005: 73(1): 1-7.
20. Omran MAA. Cytotoxic and apoptotic effects of scorpion Leiurus quinquestriatus venom on 293T and C2C12 eukaryotic cell lines. Journal of venomous animals and toxins including tropical diseases. 2003: 9(2): 255-276.
21. Fu Y J, Yin LT, Liang AH, Zhang CF, Wang W, Chai BF, ... & Fan X.J. Therapeutic potential of chlorotoxin-like neurotoxin from the Chinese scorpion for human gliomas. Neuroscience letters. 2007; 412(1): 62-67.
22. Sariego J. Breast cancer in the young patient. The American surgeon. 2010: 76(12): 1397-1400.
23. D’Suze G, Rosales A, Salazar V, Sevcik C. Apoptogenic peptides from Tityus discrepans scorpion venom acting against the SKBR3 breast cancer cell line. Toxicon. 2010; 56(8): 1497-1505.
24. Ortiz E, Gurrola GB, Schwartz EF, Possani LD. Scorpion venom components as potential candidates for drug development. Toxicon. 2015; 93: 125-135.
25. Zhang YY, Wu LC, Wang ZP, Wang ZX, Jia Q, Jiang GS, Zhang WD. Anti-proliferation effect of polypeptide extracted from scorpion venom on human prostate cancer cells in vitro. Journal of clinical medicine research. 2009; 1(1): 24-31.
26. Incesu Z, Caliskan F, Zeytinoglu H. (2005). Cytotoxic and gelatinolytic activities of Mesobuthus gibbosus (Brullé, 1832) venom. Revista CENIC. Ciencias Biológicas. 36.
27. Zargan J, Umar S, Sajad M, Naime M, Ali S, Khan HA. Scorpion venom (Odontobuthus doriae) induces apoptosis by depolarization of mitochondria and reduces S-phase population in human breast cancer cells (MCF-7). Toxicology in Vitro. 2011; 25(8): 1748-1756.