Mesobuthus eupeus venom induced injury in the colorectal carcinoma cell line (HT29) through altering the mitochondria membrane stability | ||
| Iranian Journal of Basic Medical Sciences | ||
| مقاله 8، دوره 23، شماره 6، شهریور 2020، صفحه 760-767 اصل مقاله (697.97 K) | ||
| نوع مقاله: Original Article | ||
| شناسه دیجیتال (DOI): 10.22038/ijbms.2020.40884.9659 | ||
| نویسندگان | ||
| Massood Valizade1، 2؛ Atefeh Raesi Vanani2، 3؛ Mohsen Rezaei2، 4؛ Layasadat Khorsandi1، 5؛ Leila Zeidooni2؛ Masoud Mahdavinia* 2، 3 | ||
| 1Cell & Molecular Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran | ||
| 2Toxicology Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran | ||
| 3Department of Toxicology, School of Pharmacy, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran | ||
| 4Department of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran | ||
| 5Department of Anatomical Sciences, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran | ||
| چکیده | ||
| Objective(s): The purpose of this study was to investigate cytotoxicity and membrane toxicity effects induced by Mesobuthus eupeus venom (MEV) on the HT-29 cell line. Materials and Methods: To determine the in vitro cytotoxicity via MTT assays, HT-29 (as cancer cell line) and Hek-293T (as normal cell) were treated through different concentrations of MEV, and cytotoxicity effects were then measured through assessment of mitochondrial membrane potential (ΔΨm), reactive oxygen species (ROS) generation, and apoptosis induction. The colony formation assay was performed to measure the antiproliferative effect of MEV on HT-29 cells. Nuclei alterations were also observed during apoptosis following DAPI staining. Besides, atomic force microscopy (AFM) was used to detect alterations in morphology and ultrastructure of the cells at a nanoscale level. Results: According to MTT and clonogenic assays, MEV caused a significant decrease in cell viability and proliferation of HT-29 cells while it did not have any impact on normal cells and the IC50 value was found to be 10 µg/ml. Induction of apoptosis was also confirmed by flowcytometric analysis in HT-29 cells. Moreover, the results indicated that MEV had led to a suppression of proliferation and induction of apoptosis through increased ROS and depolarization of mitochondria. Furthermore, AFM imaging demonstrated apoptosis cell death after being treated with MEV in HT-29 cells. Conclusion: This study showed that MEV had an antiproliferative effect on HT-29 cells by inducing apoptosis through the mitochondria signaling pathway. These findings suggested that MEV could be used as a promising natural remedy for cancer treatment. | ||
| کلیدواژهها | ||
| AFM؛ Apoptosis؛ HT-29 cancer cell؛ Mesobuthus eupeus venom؛ Mitochondrial membrane potential؛ ROS | ||
| مراجع | ||
|
1. Labianca R, Nordlinger B, Beretta G, Mosconi S, Mandalà M, Cervantes A, et al. Early colon cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 2013; 24:vi64-vi72. 2. Díaz-García A, Morier-Díaz L, Frión-Herrera Y, Rodríguez-Sánchez H, Caballero-Lorenzo Y, Mendoza-Llanes D, et al. anticancer effect of venom from Cuban scorpion Rhopalurus junceus against a panel of human cancer cell lines. J venom Res 2013; 4:5. 3. Khorsandi L, Saki G, Bavarsad N, Mombeini M. Silymarin induces a multi-targeted cell death process in the human colon cancer cell line HT-29. Biomed Pharmacother 2017; 94:890-897. 4. Ding J, Chua P-J, Bay B-H, Gopalakrishnakone P. Scorpion venoms as a potential source of novel cancer therapeutic compounds. Exp Biol Med 2014; 239:387-393. 5. Ortiz E, Gurrola GB, Schwartz EF, Possani LD. Scorpion venom components as potential candidates for drug development. Toxicon 2015; 93:125-135. 6. Hossain GS, Li J, Shin H-d, Du G, Liu L, Chen J. L-Amino acid oxidases from microbial sources: types, properties, functions, and applications. Appl Microbiol Biotechnol 2014; 98:1507-1515. 7. Gallagher JD, Fay MJ, North WG, McCann FV. Ionic signals in T47D human breast cancer cells. Cell Signal 1996; 8:279-284. 8. Kuhn-Nentwig L. Antimicrobial and cytolytic peptides of venomous arthropods. Cell Mol Life Sci 2003; 60:2651-2668. 9. Gupta SD, Gomes A, Debnath A, Saha A, Gomes A. Apoptosis induction in human leukemic cells by a novel protein Bengalin, isolated from Indian black scorpion venom: through mitochondrial pathway and inhibition of heat shock proteins. Chem Biol Interact 2010; 183:293-303. 10. Omran M. Cytotoxic and apoptotic effects of scorpion Leiurus quinquestriatus venom on 293T and C2C12 eukaryotic cell lines. J Venom Anim Toxins incl Trop Dis 2003; 9:255-276. 11. Omran MAA. ln vitro anticancer effect of scorpion Leiurus quinquestriatus and egyptian cobra venom. J Med Sci 2003; 3:66-86. 12. Ozkan O, Kat I. Mesobuthus eupeus scorpionism in Sanliurfa region of Turkey. J Venom Anim Toxins incl Trop Dis 2005; 11:479-491. 13. Moghimipour E, Rezaei M, Ramezani Z, Kouchak M, Amini M, Angali KA, et al. Transferrin targeted liposomal 5-fluorouracil induced apoptosis via mitochondria signaling pathway in cancer cells. Life Sci 2018; 194:104-110. 14. Ly JD, Grubb D, Lawen A. The mitochondrial membrane potential (Δψm) in apoptosis; an update. Apoptosis 2003; 8:115-128. 15. Wang J, Yi J. Cancer cell killing via ROS: to increase or decrease, that is the question. Cancer Biol Ther 2008; 7:1875-1884. 16. Muller DJ. AFM: a nanotool in membrane biology. Biochemistry 2008; 47:7986-7998. 17. Borges A, Silva S, den Camp HJO, Velasco E, Alvarez M, Alfonzo MJ, et al. In vitro leishmanicidal activity of Tityus discrepans scorpion venom. Parasitol Res 2006; 99:167-173. 18. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248-254. 19. Zargan J, Sajad M, Umar S, Naime M, Ali S, Khan HA. Scorpion (Odontobuthus doriae) venom induces apoptosis and inhibits DNA synthesis in human neuroblastoma cells. Mol Cell Biochem 2011; 348:173-181. 20. Wang W-X, Ji Y-H. Scorpion venom induces glioma cell apoptosis in vivo and inhibits glioma tumor growth in vitro. J Neurooncol 2005; 73:1-7. 21. Wojtala A, Bonora M, Malinska D, Pinton P, Duszynski J, Wieckowski MR. Methods to monitor ROS production by fluorescence microscopy and fluorometry. Methods Enzymol 2014; 542:243-262. 22. Ling L, Tan K, Lin H, Chiu G. The role of reactive oxygen species and autophagy in safingol-induced cell death. Cell Death Dis 2011; 2:e129. 23. Hori T, Kondo T, Tabuchi Y, Takasaki I, Zhao Q-L, Kanamori M, et al. Molecular mechanism of apoptosis and gene expressions in human lymphoma U937 cells treated with anisomycin. Chem Biol Interact 2008; 172:125-140. 24. Pendergrass W, Wolf N, Poot M. Efficacy of MitoTracker Green™ and CMXrosamine to measure changes in mitochondrial membrane potentials in living cells and tissues. Cytometry A 2004; 61:162-169. 25. Orazizadeh M, Khodadadi A, Bayati V, Saremy S, Farasat M, Khorsandi L. In vitro toxic effects of zinc oxide nanoparticles on rat adipose tissue-derived mesenchymal stem cells. Cell J (Yakhteh) 2015; 17:412-421. 26. Franken NA, Rodermond HM, Stap J, Haveman J, Van Bree C. Clonogenic assay of cells in vitro. Nat Protoc 2006; 1:2315-2319. 27. Vyas VK, Brahmbhatt K, Bhatt H, Parmar U. Therapeutic potential of snake venom in cancer therapy: current perspectives. Asian Pac J Trop Biomed 2013; 3:156-162. 28. Staudacher I, Jehle J, Staudacher K, Pledl HW, Lemke D, Schweizer PA, et al. HERG K+ channel-dependent apoptosis and cell cycle arrest in human glioblastoma cells. PLoS One 2014; 9:e88164. 29. Storz P. Reactive oxygen species in tumor progression. Front Biosci 2005; 10:1881-1896. 30. Georgiev K, Georgieva M, Iliev I, Peneva M, Alexandrov G. Antiproliferative effect of Bulgarian spring water probiotics (Lakter a Nature Probiotics®), against human colon carcinoma cell line. World J Pharm Pharm Sci 2015; 4:130-136. 31. Gomes A, Bhattacharjee P, Mishra R, Biswas AK, Dasgupta SC, Giri B, et al. Anticancer potential of animal venoms and toxins. Indian J Exp Biol 2010; 48:93-103. 32. Gogvadze V, Zhivotovsky B, Orrenius S. The Warburg effect and mitochondrial stability in cancer cells Mol Aspects Med 2010; 31:60-74. 33. Munshi A, Hobbs M, Meyn RE. Clonogenic cell survival assay. Chemosensitivity 2005; p. 21-28. 34. Brown JM, Attardi LD. The role of apoptosis in cancer development and treatment response. Nat Rev Cancer 2005; 5:231-237. 35. Hessler JA, Budor A, Putchakayala K, Mecke A, Rieger D, Banaszak Holl MM, et al. Atomic force microscopy study of early morphological changes during apoptosis. Langmuir 2005; 21:9280-9286. 36. Jin H, Zhong X, Wang Z, Huang X, Ye H, Ma S, et al. Sonodynamic effects of hematoporphyrin monomethyl ether on CNE‐2 cells detected by atomic force microscopy. J Cell Biochem 2011; 112:169-178. 37. Wang M, Ruan Y, Chen Q, Li S, Wang Q, Cai J. Curcumin induced HepG2 cell apoptosis-associated mitochondrial membrane potential and intracellular free Ca2+ concentration. Eur J Pharmacol 2011; 650:41-47. 38. Le Grimellec C, Lesniewska E, Giocondi M-C, Finot E, Vié V, Goudonnet J-P. Imaging of the surface of living cells by low-force contact-mode atomic force microscopy. Biophys 1998; 75:695-703. 39. Lansu K, Gentile S. Potassium channel activation inhibits proliferation of breast cancer cells by activating a senescence program. Cell Death Dis 2013; 4:e652. 40. Huang X, Jan LY. Targeting potassium channels in cancer. J Cell Biol 2014; 206:151-162. 41. Melo ETd. Avaliação funcional e estrutural de um novo peptídeo antimicrobiano do escorpião Tityus stigmurus: UFRN 2014. 42. Su JC, Lin K, Wang Y, Sui SH, Gao ZY, Wang ZG. In vitro studies of phenethyl isothiocyanate against the growth of LN229 human glioma cells. Int J Clin Exp Pathol 2015; 8:4269-4276. | ||
|
آمار تعداد مشاهده مقاله: 771 تعداد دریافت فایل اصل مقاله: 445 |
||