نوع مقاله : مقاله پژوهشی
نویسندگان
1 دانشجوی دکترای زیست فناوری، بخش بیوتکنولوژی، دانشگاه آزاد اسالمی واحد ساوه، ساوه ایران
2 استاد تمام سلولی و مولکولی، بخش بیوتکنولوژی، دانشگاه آزاد اسالمی واحد ساوه، ساوه ایران
چکیده
زمینه و هدف: امروزه، نانو ذرات بهعنوان یک فاکتور ضد سرطانی در تحقیقات حوزه درمان سرطان موردتوجه قرارگرفته و هدف از مطالعه حاضر تولید نانو ذره اکسید روی از بیوفیلم لاکتوباسیلوس پلانتاروم و تأثیر آن بر بیان ژنهای P53 و NF-kB در سلولهای اپیتلیال سرطان روده بزرگ است.
مواد و روشها: پس از جداسازی و تائید نانو ذرات اکسید روی از بیوفیلم لاکتوباسیلوس پلانتاروم سلولهای HT-29 با غلظتهای مختلف نانو ذرات اکسید روی به مدت 24 ساعت کشت داده شد.
در این مطالعه، پس از بیوسنتز نانو ذرات ZnO توسط لاکتوباسیلوس پلانتاروم، شناسایی نانو ذرات توسط میکروسکوپ الکترونی روبشی UV-Vis، XRD و SEM انجام شد. پسازآنجام آزمایش MTT غلظت موردنظر انتخاب و سپس بیان P53 و NF-kB در سطح mRNA با استفاده از پرایمرهای جدید طراحیشده از روش Reverse transcription (RT) qPCR انجام و با نرمافزار GeniX6، مورد آنالیز قرار گرفت.
یافتهها: نتایج آزمایش MTT نشان داد که غلظت 40 µg/ml بیشترین کشندگی طی 24 ساعت را دارد. پس از 24 ساعت انکوباسیون با نانوذره اکسید روی، میزان بیان ژن P53 به میزان معنیداری در گروه تحت درمان نسبت به گروه کنترل افزایش یافت (P<0.0001)؛ اما میزان بیان NF-kB در گروه تحت درمان با µg/ml10 نسبت به گروه کنترل کاهش معنیداری را نشان داد (P < 0.001). همچنین تعداد سلولهای موردبررسی در گروه تحت تیمار نسبت به گروه کنترل کاهش معنیداری را نشان داد (P<0.0001).
نتیجهگیری: : نانو ذرات اکسید روی باعث ایجاد مرگ در سلولهای HT-29 میشوند. همچنین نانو اکسید روی موجب افزایش بیان ژن P53 و کاهش بیان ژن NF-kB در سلولهای سرطانی میشود. نتایج مطالعه حاضر میتواند زمینه را برای تحقیقات بیشتر و استفاده از نانو ذرات اکسید روی در صنایع دارویی و درمان سرطان روده بزرگ فراهم سازد.
تازه های تحقیق
https://scholar.google.com/citations?user=Au7Bt6YAAAAJ&hl=en
https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography
https://scholar.google.com/citations?user=4KmKeNsAAAAJ&hl=en&oi=ao
https://pubmed.ncbi.nlm.nih.gov/33753577/
https://scholar.google.com/citations?user=DMiK1IMAAAAJ&hl=en&oi=ao
https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/
https://scholar.google.com/citations?view%20op=list%20works&hl=en&hl=en&user=6BZaLvIAAAAJ
https://pubmed.ncbi.nlm.nih.gov/35424885/
کلیدواژهها
موضوعات
عنوان مقاله [English]
Effects of Zinc Oxide (ZnO) Nanoparticles Extracted from Lactobacillus plantarum on mRNA Levels of P53 and NF-κB Genes in Colorectal Cancer Epithelial Cells
نویسندگان [English]
- Mohammadreza Azimi 1
- Anoush Eghdami 2
- Bahram Keyvani 2
- Ramin Cheragh-Ali 2
1 PhD Student, Department of Biochemistry, Medical Faculty, Saveh Branch, Islamic Azad University, Saveh, Iran
2 Professor, Department of Biochemistry, Medical Faculty, Saveh Branch, Islamic Azad University, Saveh, Iran
چکیده [English]
Introduction: Nanoparticles are increasingly recognized as potential anticancer agents in cancer treatment research. This study aimed to synthesize zinc oxide nanoparticles from Lactobacillus plantarum biofilms and assess their impact on the expression of the P53 and NF-κB genes in colorectal cancer epithelial cells.
Materials and Methods: Following the confirmation and isolation of zinc oxide nanoparticles from Lactobacillus plantarum biofilm, HT-29 cells were cultured with varying concentrations of these nanoparticles for 24 hours. The MTT assay was utilized to evaluate the toxicity of the isolated nanoparticles. The optimal concentration was determined based on these results, after which the expression levels of P53 and NF-κB mRNA were analyzed.
Results: The MTT assay indicated that a concentration of 40 µg/ml had the highest lethality after 24 hours. Notably, P53 expression significantly increased in the treatment group compared to the control group (P < 0.0001), while NF-κB expression significantly decreased at a concentration of 10 µg/ml (P < 0.001). Furthermore, the cell count in the treated group showed a significant reduction relative to the control group (P < 0.0001)
Conclusion: : ZnO nanoparticles induce lethality in HT-29 cells and enhance P53 expression, thereby suppressing tumor cell proliferation. These findings provide a foundation for further research and underscore the potential of zinc oxide nanoparticles in the pharmaceutical industry and colorectal cancer treatment
کلیدواژهها [English]
- Colorectal cancer
- Zinc Oxide (ZnO) Nanoparticles
- MTT assay
- Quantitative Polymerase Chain Reaction (qPCR)
- P53
- NF-κB
- Azimi M, Mehrzad J, Ahmadi A, Ahmadi E, Ghorbani Ranjbary A. Apoptosis Induced by Ziziphora tenuior Essential Oil in Human Colorectal Cancer Cells. Biomed Res Int. 2021 Jul 22;2021:5522964. doi: 10.1155/2021/5522964.
- Ranjbary AG, Mehrzad J, Dehghani H, Abdollahi A, Hosseinkhani S. Variation in blood and colorectal epithelia’s key trace elements along with expression of mismatch repair proteins from localized and metastatic colorectal cancer patients. Biological trace element research. 2020 Mar;194(1):66-75. doi: 10.1007/s12011-019-01749-9.
- Ranjbary AG, Saleh GK, Azimi M, Karimian F, Mehrzad J, Zohdi J. Superparamagnetic Iron Oxide Nanoparticles Induce Apoptosis in HT-29 Cells by Stimulating Oxidative Stress and Damaging DNA. Biological Trace Element Research. 2022 Apr 22:1-1. doi: 10.1007/s12011-022-03229-z.
- Nagai H, Kim YH. Cancer prevention from the perspective of global cancer burden patterns. J Thorac Dis. 2017 Mar;9(3):448-451. doi: 10.21037/jtd.2017.02.75.
- ReFaey K, Tripathi S, Grewal SS, Bhargav AG, Quinones DJ, Chaichana KL, Antwi SO, Cooper LT, Meyer FB, Dronca RS, Diasio RB, Quinones-Hinojosa A. Cancer Mortality Rates Increasing vs Cardiovascular Disease Mortality Decreasing in the World: Future Implications. Mayo Clin Proc Innov Qual Outcomes. 2021 Jun 8;5(3):645-653. doi: 10.1016/j.mayocpiqo.2021.05.005.
- Rawla P, Sunkara T, Barsouk A. Epidemiology of colorectal cancer: incidence, mortality, survival, and risk factors. Prz Gastroenterol. 2019;14(2):89-103. doi: 10.5114/pg.2018.81072.
- Xi Y, Xu P. Global colorectal cancer burden in 2020 and projections to 2040. Transl Oncol. 2021 Oct;14(10):101174. doi: 10.1016/j.tranon.2021.101174.
- Baskar R, Lee KA, Yeo R, Yeoh KW. Cancer and radiation therapy: current advances and future directions. Int J Med Sci. 2012;9(3):193-9. doi: 10.7150/ijms.3635.
- Ranjbary AG, Bagherzadeh A, Sabbaghi SS, Faghihi A, Karimi DN, Naji S, Kardani M. Chlorogenic acid induces apoptosis and cell-cycle arrest in colorectal cancer cells. Mol Biol Rep. 2023 Dec;50(12):9845-9857. doi: 10.1007/s11033-023-08854-y.
- Ghorbani Ranjbary A, Mehrzad J, Rahbar N, Dehghani H. Impacts of some clinicopathodemography and colorectal tissues key cell cycle and mucin stabilizing molecules on the metastasis trend in colorectal cancer patients. Mol Biol Rep. 2023 Oct;50(10):8589-8601. doi: 10.1007/s11033-023-08766-x.
- Rasmussen JW, Martinez E, Louka P, Wingett DG. Zinc oxide nanoparticles for selective destruction of tumor cells and potential for drug delivery applications. Expert Opin Drug Deliv. 2010 Sep;7(9):1063-77. doi: 10.1517/17425247.2010.502560.
- Bisht G, Rayamajhi S. ZnO Nanoparticles: A Promising Anticancer Agent. Nanobiomedicine (Rij). 2016 Jan 1;3:9. doi: 10.5772/63437.
- Tanino R, Amano Y, Tong X, Sun R, Tsubata Y, Harada M, Fujita Y, Isobe T. Anticancer Activity of ZnO Nanoparticles against Human Small-Cell Lung Cancer in an Orthotopic Mouse ModelZnO Nanoparticles Inhibit Growth of Small-Cell Lung Cancer. Molecular Cancer Therapeutics. 2020 Feb 1;19(2):502-12. doi: 10.1158/1535-7163.
- Yousef Aljohar A, Muteeb G, Zia Q, AATIF M, Farhan M, Khan M, Alsultan A, Jamal A, Alshoaibi A, Ahmad E, Alam W. Anticancer effect of zinc oxide nanoparticles prepared at diverse entry time of ion carriers against A431 skin cancer cells in vitro. Frontiers in Chemistry.:1510. doi: 10.3389/fchem.2022.1069450.
- Wilhelmi V, Fischer U, Weighardt H, Schulze-Osthoff K, Nickel C, Stahlmecke B, Kuhlbusch TA, Scherbart AM, Esser C, Schins RP, Albrecht C. Zinc oxide nanoparticles induce necrosis and apoptosis in macrophages in a p47phox- and Nrf2-independent manner. PLoS One. 2013 Jun 3;8(6):e65704. doi: 10.1371/journal.pone.0065704.
- Chang F, Syrjänen S, Kurvinen K, Syrjänen K. The p53 tumor suppressor gene as a common cellular target in human carcinogenesis. Am J Gastroenterol. 1993 Feb;88(2):174-86. PMID: 8424417.
- Rivlin N, Brosh R, Oren M, Rotter V. Mutations in the p53 Tumor Suppressor Gene: Important Milestones at the Various Steps of Tumorigenesis. Genes Cancer. 2011 Apr;2(4):466-74. doi: 10.1177/1947601911408889.
- Rivlin N, Brosh R, Oren M, Rotter V. Mutations in the p53 Tumor Suppressor Gene: Important Milestones at the Various Steps of Tumorigenesis. Genes Cancer. 2011 Apr;2(4):466-74. doi: 10.1177/1947601911408889.
- Oeckinghaus A, Ghosh S. The NF-kappaB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol. 2009 Oct;1(4):a000034. doi: 10.1101/cshperspect.a000034.
- Xia Y, Shen S, Verma IM. NF-κB, an active player in human cancers. Cancer Immunol Res. 2014 Sep;2(9):823-30. doi: 10.1158/2326-6066.CIR-14-0112.
- Hayden MS, Ghosh S. NF-κB, the first quarter-century: remarkable progress and outstanding questions. Genes Dev. 2012 Feb 1;26(3):203-34. doi: 10.1101/gad.183434.111.
- Liang Y, Chen G, Yang Y, Li Z, Chen T, Sun W, Yu M, Pan K, Guo W, Tian W. Effect of canonical NF-κB signaling pathway on the differentiation of rat dental epithelial stem cells. Stem Cell Research & Therapy. 2019 Dec;10(1):1-2. doi: 10.1186/s13287-019-1252-7.
- Ghorbani Ranjbary A, Mehrzad J, Dehghani H, Abdollahi A, Hosseinkhani S. The study of expression of KI67 and P53 in co-culturing of colon epithelial cell line HT-29 with IL-17A. Journal of Sabzevar University of Medical Sciences. 2022 May 18;29(1):141-53.
- L SW, Lee CH, Lin MS, Chi CW, Chen YJ, Wang GS, Liao KW, Chiu LP, Wu SH, Huang DM, Chen L, Shen YS. ZnO Nanoparticles Induced Caspase-Dependent Apoptosis in Gingival Squamous Cell Carcinoma through Mitochondrial Dysfunction and p70S6K Signaling Pathway. Int J Mol Sci. 2020 Feb 26;21(5):1612. doi: 10.3390/ijms21051612.
- Anjum S, Hashim M, Malik SA, Khan M, Lorenzo JM, Abbasi BH, Hano C. Recent Advances in Zinc Oxide Nanoparticles (ZnO NPs) for Cancer Diagnosis, Target Drug Delivery, and Treatment. Cancers (Basel). 2021 Sep 12;13(18):4570. doi: 10.3390/cancers13184570.
- Bai DP, Zhang XF, Zhang GL, Huang YF, Gurunathan S. Zinc oxide nanoparticles induce apoptosis and autophagy in human ovarian cancer cells. Int J Nanomedicine. 2017 Sep 5;12:6521-6535. doi: 10.2147/IJN.S140071.
- Sadeghzadeh F, Golestani P, Beyramabdi P, Pouresmaeil V, Hosseini H, Homayouni Tabrizi M. The anticancer impact of folate-linked ZnO-decorated bovine serum albumin/silibinin nanoparticles on human pancreatic, breast, lung, and colon cancers. J Biomater Sci Polym Ed. 2024 May 29:1-18. doi: 10.1080/09205063.2024.2356967.
- Akhtar MJ, Ahamed M, Kumar S, Khan MM, Ahmad J, Alrokayan SA. Zinc oxide nanoparticles selectively induce apoptosis in human cancer cells through reactive oxygen species. Int J Nanomedicine. 2012;7:845-57. doi: 10.2147/IJN.S29129.
- Anjum S, Hashim M, Malik SA, Khan M, Lorenzo JM, Abbasi BH, Hano C. Recent advances in zinc oxide nanoparticles (Zno nps) for cancer diagnosis, target drug delivery, and treatment. Cancers. 2021 Sep 12;13(18):4570. doi: 10.3390/cancers13184570.
- Kim MH, Jeong HJ. Zinc Oxide Nanoparticles Suppress LPS-Induced NF-κB Activation by Inducing A20, a Negative Regulator of NF-κB, in RAW 264.7 Macrophages. J Nanosci Nanotechnol. 2015 Sep;15(9):6509-15. doi: 10.1166/jnn.2015.10319.
- Deylam M, Alizadeh E, Sarikhani M, Hejazy M, Firouzamandi M. Zinc oxide nanoparticles promote the aging process in a size-dependent manner. Journal of materials science: Materials in medicine. 2021 Oct;32(10):1-0. doi: 10.1007/s10856-021-06602-x.
- Wahab R, Siddiqui MA, Saquib Q, Dwivedi S, Ahmad J, Musarrat J, Al-Khedhairy AA, Shin HS. ZnO nanoparticles induced oxidative stress and apoptosis in HepG2 and MCF-7 cancer cells and their antibacterial activity. Colloids Surf B Biointerfaces. 2014 May 1;117:267-76. doi: 10.1016/j.colsurfb.2014.02.038.
- Subramaniam VD, Ramachandran M, Marotta F, Banerjee A, Sun XF, Pathak S. Comparative study on anti-proliferative potentials of zinc oxide and aluminium oxide nanoparticles in colon cancer cells. Acta Biomed. 2019 May 23;90(2):241-247. doi: 10.23750/abm.v90i2.6939.
- Siddiqi KS, Husen A. Properties of zinc oxide nanoparticles and their activity against microbes. Nanoscale research letters. 2018 Dec;13(1):1-3. doi: 10.1186/s11671-018-2532-3.
- Liao C, Jin Y, Li Y, Tjong SC. Interactions of zinc oxide nanostructures with mammalian cells: cytotoxicity and photocatalytic toxicity. International Journal of Molecular Sciences. 2020 Aug 31;21(17):6305. doi: 10.3390/ijms21176305.
- Sirelkhatim A, Mahmud S, Seeni A, Kaus NH, Ann LC, Bakhori SK, Hasan H, Mohamad D. Review on zinc oxide nanoparticles: antibacterial activity and toxicity mechanism. Nano-micro letters. 2015 Jul;7(3):219-42. doi: 10.1007/s40820-015-0040-x