اثر نانو داروی فلورسنت تاموکسیفن بر بیان ژن های مسیر آپوپتوز و چرخه سلولی در سلولهای سرطان پستان MCF-7
محورهای موضوعی : فصلنامه زیست شناسی جانوری
نورا امرایی
1
,
مریم بی خوف تربتی
2
,
احمد مجد
3
,
مسعود شعبانزاده
4
1 - گروه زیست شناسی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران
2 - گروه زیست شناسی، واحد یادگار امام خمینی (ره) شهرری، دانشگاه آزاد اسلامی، تهران، ایران
3 - گروه زیست شناسی، واحد تهران شمال، دانشگاه آزاد اسلامی، تهران، ایران
4 - گروه شیمی، واحد دامغان، دانشگاه آزاد اسلامی، دامغان، ایران
کلید واژه: تاموکسیفن, GQDs, Real-Time PCR, دارورسانی, سرطان پستان, MCF-7.,
چکیده مقاله :
تاموکسیفن رایج ترین داروی درمان سرطان پستان با گیرنده استروژن مثبت (ER+) می باشد، اثرات آنتاگونیست تاموکسیفن علیه ERs مانع از سنتز DNA و پاسخ سلولی سلولهای سرطانی به اثرات تحریکی استروژن میشود و در نتیجه مرگ سلولی را افزایش میدهد. به منظور غلبه بر عوارض تاموکسیفن و حلالیت کم و ناچیز این دارو، سیستمهای بسیاری با قابلیت نانو برای حمل مولکولهای تاموکسیفن توسعه یافتهاند که توانایی انتقال تاموکسیفن با دوز کمتر و ایمنتر به تومورها و محافظت از مولکولهای آبگریز تاموکسیفن در برابر تخریب توسط ماکروفاژها در جریان خون را دارند. اثر نانو داروی فلورسنت تاموکسیفن TMX-FPGs)) روی سلولهای سرطانی MCF-7 با روش MTT مورد ارزیابی قرار گرفت. تغییر بیان ژنهایP53 ،BCL2 ،BAX وCyclinD1 در سلولهای MCF-7تیمار شده با نانو دارو با استفاده از Real-Time PCR بررسی شد. همچنین جذب سلولی نانودارو با میکروسکوپ کانفوکال مورد ارزیابی قرار گرفت. نانو داروی TMX-FPGs با روشی وابسته به غلظت سبب کاهش توان زیستی سلولهای MCF-7 گردید و با افزایش بیان P53 و نسبت BAX/BCL2 و کاهش بیان CyclinD1 موجب افزایش معنیدار فعالیت آپوپتوز شد. همچنین، جذب سلولی و نشر فلورسانس نانودارو در تصویربرداری سلولی با میکروسکوپ کانفوکال تایید شد. با توجه به خواص فلورسانس نانودارو و توانایی آن در القاء آپوپتوز با افزایش بیان ژنهای دخیل در این امر و اثر سیتوتوکسیک آن بر سلولهای سرطانی MCF-7، نانو داروی TMX-FPGs کاندید مناسبی برای نظارت و هدفگیری سلولهای سرطانی MCF-7 میباشد.
Tamoxifen is the most common drug for the treatment of estrogen receptor-positive (ER+) breast cancer. The antagonistic effects of tamoxifen against ERs prevent DNA synthesis and the cellular response of cancer cells to the stimulating effects of estrogen, and as a result, increase cell death. In order to overcome the side effects of tamoxifen and the low solubility of this drug, many nanosystems have been developed to carry tamoxifen molecules, which have the ability to deliver tamoxifen at a lower and safer dose to tumors and protect the hydrophobic molecules of tamoxifen against destruction by macrophages in the bloodstream. The effect of fluorescent nano drug tamoxifen (TMX-FPGs) on MCF-7 cancer cells was evaluated by the MTT method. The expression changes of P53, BCL2, BAX, and CyclinD1 genes in MCF-7 cells treated with nano drugs were investigated using Real-Time PCR. Also, cellular uptake of the nano drug was evaluated using confocal microscopy. TMX-FPGs nano drug decreased the viability of MCF-7 cells in a dose-dependent manner, and with the increase of P53 expression and BAX/BCL2 ratio, and the decrease of CyclinD1 expression, significantly increased apoptotic activity. Also, cell uptake and fluorescence emission of the nano drug were confirmed in cell imaging with a confocal microscope. Considering the fluorescence properties of the nano drug and its ability to induce apoptosis by increasing the expression of genes involved in this, and its cytotoxic effect on MCF-7 cancer cells, the TMX-FPGs nano drug is a suitable candidate for monitoring and targeting MCF-7 cancer cells.
1. Day CM, Hickey SM, Song Y, Plush SE, Garg S. Novel tamoxifen nanoformulations for improving breast cancer treatment: Old wine in new bottles. Molecules. 2020; 25(5):1182.
2. Feng Y, Spezia M, Huang S, Yuan C, Zeng Z, Zhang L, et al. Breast cancer development and progression: Risk factors, cancer stem cells, signaling pathways, genomics, and molecular pathogenesis. Genes Dis. 2018;5(2):77-106.
3. Hung MH, Liu CJ, Teng CJ, Hu YW, Yeh CM, Chen SC, et al. Risk of second non-breast primary cancer in male and female breast cancer patients: a population-based cohort study. PLoS One. 2016;11(2): e0148597.
4. Yip CH, Rhodes A. Estrogen and progesterone receptors in breast cancer. Future Oncol. 2014;10(14):2293-2301.
5. Kim N, Lukong KE. Treating ER-positive breast cancer: a review of the current FDA-approved SERMs and SERDs and their mechanisms of action. Front Oncol Rev. 2025; 15:1564642.
6. Linowiecka K, Szpotan J, Godlewska M, Gaweł D, Zarakowska E, Gackowski D, et al. Selective estrogen receptor modulators’ (SERMs) influence on TET3 expression in breast cancer cell lines with distinct biological subtypes. Int J Mol Sci. 2024;25(16):8561.
7. Jena SK, Sangamwar AT. Polymeric micelles: a promising tool for tamoxifen delivery in cancer? Ther. Deliv. 2017;8(3): 109-111.
8. Öztürk-Atar K, Kaplan M, Çalış S. Development and evaluation of polymeric micelle containing tablet formulation for poorly water-soluble drug: tamoxifen citrate. Drug Dev Ind Pharm. 2020; 46(10):1695-1704.
9. Xuan QJ, Wang JX, Nanding A, Wang ZP, Liu H, Lian X, Zhang QY. Tumor-associated macrophages are correlated with tamoxifen resistance in the postmenopausal breast cancer patients. Pathol Oncol Res. 2014;20:619-624.
10. Ventola CL. Progress in nanomedicine: approved and investigational nanodrugs. P&T. 2017;42(12):742.
11. Farjadian F, Ghasemi A, Gohari O, Roointan A, Karimi M, Hamblin MR. Nanomedicine and drug delivery: A comprehensive review of recent advances. Nanomedicine. 2019;14(1):93–126.
12. Chung S, Revia RA, Zhang M. Graphene quantum dots and their applications in bioimaging, biosensing, and therapy. Adv. Mater. 2021;33(22):1904362.
13. Some S, Gwon AR, Hwang E, Bahn GH, Yoon Y, Kim Y, Kim SH, Bak S, Yang J, Jo DG, Lee H. Cancer therapy using ultrahigh hydrophobic drug-loaded graphene derivatives. Sci Rep. 2014;4(1):6314.
14. Eanes LA, Eldeeb M, Storholt D, Patel YM. Naringenin impairs mitochondrial function via ROS to induce apoptosis in tamoxifen-resistant MCF-7 breast cancer cells. PLoS One. 2025;20(4):e0320020.
15. Jayaraman S, Wu X, Kalari KR, Tang X, Kuffel MJ, Bruinsma ES, et al. Endoxifen downregulates AKT phosphorylation through protein kinase C beta 1 inhibition in ERα⁺ breast cancer. NPJ Breast Cancer. 2023;9:101.
16. Akbarzadeh I, Farid M, Javidfar M, Zabet N, Shokoohian B, Kazem Arki M, et al. The optimized formulation of tamoxifen-loaded niosomes efficiently induced apoptosis and cell cycle arrest in breast cancer cells. AAPS Pharm Sci Tech. 2022; 23(2):56.
17. Khan R, Qureshi A, Azhar M, et al. Recent progress of fluorescent carbon dots and graphene quantum dots for biosensors: synthesis of solution methods and their medical applications. J Fluoresc. 2024; 34(3):765-782.
18. Zarepour A, Hasanzadeh M, Shadjou N. Innovative approaches for cancer treatment: graphene quantum dots for photodynamic and photothermal therapies. J Mater Chem B. 2024;12(5):1021-1035.
19. Rouhimoghadam M, Safarian S, Carroll JS, Sheibani N, Bidkhori G. Tamoxifen-induced apoptosis of MCF-7 cells via GPR30/PI3K/MAPKs interactions: verification by ODE modeling and RNA sequencing. Front Physiol. 2018;9:907
20. Yang T, Li W, Zhou J, Xu M, Huang Z, Ming J, Huang T. A novel bystander effect in tamoxifen treatment: PPIB derived from ER⁺ cells attenuates ER⁻ cells via endoplasmic reticulum stress-induced apoptosis. Cell Death Dis. 2024;15(2):147.
21. Abbasalipourkabir R, Salehzadeh A, Abdullah R. Tamoxifen-loaded solid lipid nanoparticles-induced apoptosis in breast cancer cell lines. J Exp Nanosci. 2016; 11(3):161-74.
22. Yang T, Li W, Zhou J, Xu M, Huang Z, Ming J, Huang T. A novel bystander effect in tamoxifen treatment: PPIB derived from ER+ cells attenuates ER−cells via endoplasmic reticulum stress-induced apoptosis. Cell Death Dis. 2024;15(2):147.
23. Dehghan MH, Hedayati M, Shivaee S, Shakib H, Rajabi S. Tamoxifen triggers apoptosis of papillary thyroid cancer cells by two different mechanisms. Gene Rep. 2021; 24:101266.
24. Montalto FI, De Amicis F. Cyclin D1 in cancer: a molecular connection for cell cycle control, adhesion and invasion in tumor and stroma. Cells. 2020;9(12):2648.
25. Hanf D, Müller V, Schmidt M, et al. Impact of CCND1 amplification on the prognosis of hormone receptor–positive, HER2-negative breast cancer patients. Breast Cancer Res Treat. 2025;194(2):245-256.
26. Erdyneeva DB, Babyshkina NN, Dronova TA, Vtorushin SV, Slonimskaya EM, Stegniy V.N., et al. Role of cyclin D1 in the mechanisms of tamoxifen resistance. Sib J Oncol. 2020;19(4):138-145.
27. Ishii Y, Waxman S, Germain D. Tamoxifen stimulates the growth of cyclin D1–overexpressing breast cancer cells by promoting the activation of signal transducer and activator of transcription 3. Cancer Res. 2008;68(3):852-860.
28. Ichikawa A, Ando J, Suda K. G1 arrest and expression of cyclin-dependent kinase inhibitors in tamoxifen-treated MCF-7 human breast cancer cells. Hum. Cell. 2008;21:28-37.
29. Wang X, Simpson ER, Brown KA. p53: protection against tumor growth beyond effects on cell cycle and apoptosis. Cancer Res. 2015;75(23):5001-7.
30. Qian S, Wei Z, Yang W, Huang J, Yang Y, Wang J. The role of BCL-2 family proteins in regulating apoptosis and cancer therapy. Front Oncol. 2022;12:985363.
31. Huang X, Zhang Q, Wang N. The significance of Bcl-2 expression in breast cancer: A 10-year follow-up study. J Clin Invest. 2021;4(1):1-6.
32. Kawiak A, Kostecka A. Regulation of Bcl-2 family proteins in estrogen receptor-positive breast cancer and their implications in endocrine therapy. Cancers. 2022;14(2): 279.
33. Martinez-Arribas F, Martín-Garabato E, Zapardiel I, Sánchez J, Lucas AR, Tejerina A, Schneider J. Bax expression in untreated breast cancer: an immunocytometric study of 255 cases. Anticancer Res. 2008;28(5A): 2595-2598.
34. Kim IY, Han SY, Moon A. Phthalates inhibit tamoxifen-induced apoptosis in MCF-7 human breast cancer cells. J. Toxicol. Environ. Health A. 2004;67(23-24):2025-2035.
35. Kilker RL, Planas-Silva MD. Cyclin D1 is necessary for tamoxifen-induced cell cycle progression in human breast cancer cells. Cancer Res. 2006;66(23):11478-11484.
36. Zhang Y, Wang Y, Li X. Cyclin D1 suppression enhances tamoxifen sensitivity by arresting cell cycle progression in ER-positive breast cancer. Mol Cancer Ther. 2023;22(4):512–524.
37. Shi Q, Li Y, Jin L, et al. LncRNA DILA1 inhibits Cyclin D1 degradation and contributes to tamoxifen resistance in breast cancer. Nat Commun. 2020;11:5515.
