یافته های نوین در علوم زیستی

اثر اگزوزوم‌های مشتق از سلول‌های سرطانی اپیتلیالی تخمدان انسانی بر ترشح هورمون‌های AMH و Inhibin سلول‌های گرانولوزا

نویسندگان

نجمه نیکدل 1
جواد بهارآرا 2
سعید ذاکر بستان آباد 3
مریم طهرانی پور 1

1 گروه زیست شناسی، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران

2 گروه زیست شناسی، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران؛ مرکز تحقیقات بیولوژی کاربردی تکوین جانوری، واحد مشهد، دانشگاه آزاد اسلامی، مشهد، ایران

3 گروه میکروب شناسی، واحد پرند، دانشگاه آزاد اسلامی، تهران، ایران

https://doi.org/10.52547/nbr.8.1.31
چکیده
اگزوزوم از انواع مختلف سلول­ها ترشح می­شود و به­عنوان بسته­های زیستی شناخته می­شوند. اگزوزوم­ها در ارتباطات بین سلولی و همچنین در ایجاد و پیشرفت بیماری­های مختلف از جمله سرطان نقش مهمی دارند. هورمون­های Inhibin و آنتی­مولرین هورمون (AMH) از نشانگرهای تومور سلول گرانولوزا (GCT) است. در این مطالعه تجربی آزمایشگاهی اثر اگزوزوم­های مشتق از سلول­های سرطان تخمدان انسانی بر ترشح هورمون Inhibin B و AMH از سلول­های گرانولوزا بررسی شد. ابتدا سلول­های رده A2780 سرطان تخمدان کشت داده شدند سپس مایع رویی جهت استخراج اگزوزوم­ها توسط اولتراسانتریفیوژ جمع­آوری و با استفاده از روش DLS و میکروسکوپ الکترونی روبشی (SEM) بررسی شدند. همچنین سلول­های گرانولوزا از تخمدان موش ماده نابالغ نژاد Balb/C استخراج و کشت داده شد و سپس توسط غلظت μg/ml 25 از اگزوزوم­های استخراج شده سلول­های سرطان تخمدان تیمار شد و مقدار هورمون­های Inhibin B و AMH اندازه­گیری شد. مقدار هورمون­های Inhibin B و AMH در سلول­های تیمار شده در مقایسه با شاهد افزایش معنی­دار نشان داد 05/0P<. از آنجا که در این مطالعه اگزوزوم­های استخراج شده از سلول های سرطانی تخمدان باعث افزایش معنی­دار هورمون­های Inhibin B و AMH شده است به­نظر می­رسد که در ایجاد تومور سلول گرانولوزا موثر می­باشند و لذا ضرورت مطالعات کلینیکی کاملا محسوس و پیشنهاد می­شود.



کلیدواژه‌ها

اگزوزوم
اولتراسانتریفیوژ
تومور سلول گرانولوزا
Inhibin B
AMH

عنوان مقاله English

The effect of exosomes derived from human ovarian epithelial cancer cells on the secretion of AMH and Inhibin in granulosa cells

نویسندگان English

Najme Nikdel 1
Javad Baharara 2
Saeed Zakerbostanabad 3
Maryam Tehranipour 1
1 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
2 Department of Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran; Research Center for Animal Development Applied Biology, Mashhad Branch, Islamic Azad University, Mashhad, Iran
3 Department of Microbiology, Parand Branch, Islamic Azad University, Tehran, Iran
چکیده English

Exosomes are secreted by different types of cells and known as biological packages. Exosomes have significant role in intercellular communications and involved in the development and progression of various diseases such as cancer. Inhibin B and anti-mullerian hormone (AMH) are markers of granulosa cell tumors (GCT) and due to the role of exosomes in the progression of cancer, in this experimental study, the effect of exosomes derived from human ovarian cancer cells on the secretion of Inhibin B and antimullerian hormone (AMH) by granulosa cells was investigated. First, A2780 human ovarian cancer cells were cultured, then the supernatant was collected to extract the exosomes by ultracentrifugation and subsequently, the extracted exosomes were checked out using dynamic light scattering (DLS) and Scanning electron microscopy (SEM). In addition, granulosa cells were isolated and cultured from the ovaries immature female Balb / C mice and treated with 25 μg/ml of exosomes derived from the ovarian cancer cell-line. Inhibin B and AMH hormones levels were then measured. The results showed the significant (P<0/05) increase of the level of inhibin B and AMH hormones in the treated cells in comparison with the control group. According to the results, exosomes increased the secretion of Inhibin B and AMH hormones and seems to be effective in the proliferation of granulosa cell tumors.



کلیدواژه‌ها English

exosome
ultracentrifugation
granulosa cell tumor
Inhibin B
AMH
Amini, E., Baharara, J., Nikdel, N., & Salek Abdollahi, F. 2015. Cytotoxic and pro-apoptotic effects of honey bee venom and chrysin on human ovarian cancer cells. Asia Pacific Journal of Medical Toxicology 4: 68-73.
Baharara J., Ramezani T., Saghiri N. & Salek F. 2019. Investigating the apoptotic effects of silver nanoparticles coated with Achillea biebersteinii extract on A2780 ovarian cancer cells. Nova Biologica Reperta 6: 140-147
Chuo, S.T., Chien, J.C. & Lai, C.P. 2018. Imaging extracellular vesicles: current and emerging methods. Journal of Biomedical Science 25: 1-10.
Ding, T., Luo, A., Yang, S., Lai, Z., Wang, Y., Shen, W., Jiang, J., Lu, Y., Ma, D. & Wang, S. 2012. Effects of basal media and supplements on diethylstilbestrol‐treated immature mouse primary granulosa cell growth and regulation of steroidogenesis in vitro. Reproduction in Domestic Animals 47: 355-364.
Franzen C.A., Blackwell R.H., Todorovic V., Greco K.A., Foreman K.E., Flanigan R.C., Kuo PC. & Gupta G.N. 2015. Urothelial cells undergo epithelial-to-mesenchymal transition after exposure to muscle invasive bladder cancer exosomes. Oncogenesis 4: 151-163.
Geerts I., Vergote I., Neven P., & Billen J. 2009. The role of inhibins B and antimüllerian hormone for diagnosis and follow-up of granulosa cell tumors. International Journal of Gynecological Cancer 19: 847-55.
Haltia U.M, Hallamaa M., Tapper J., Hynninen J., Alfthan H., Kalra B., Ritvos O., Heikinheimo M., Unkila-Kallio L., Perheentupa A. & Färkkilä A. 2017. Roles of human epididymis protein 4, carbohydrate antigen 125, inhibin B and anti-Müllerian hormone in the differential diagnosis and follow-up of ovarian granulosa cell tumors. Gynecologic Oncology 144: 83-89.
Hasanzadeh Mofrad, M., Hosseini Hoshyar A.S., Ghazanfari SM. & Boroumand Noghabi S. 2012. ‘Granulosa cell tumor associated with chondrosarcoma of iliac: A case report. Quarterly Journal of Sabzevar University of Medical Sciences 19: 304-309.
Healy, D.L., Burger H., Mamers P., Jobling T., Bangah M. & Quinn M. 1993. ‘Elevated serum Inhibin concentrations in postmenopausal women with ovarian tumors’. The New England Journal of Medicine 329: 1539-1542.
Kim, S.-Y., Ebbert, K., H. Cordeiro, M., Romero, M.M., Whelan, K.A. Suarez, A.A., Woodruff, T.K. & Kurita, T. 2016. Constitutive activation of PI3K in aocyte induces ovarian granulosa cell tumors. Cancer Research 76: 3851-3861.
Li, J., Bao R., Peng, S., & Zhang, C. 2018. The molecular mechanism of ovarian granulosa cell tumors. Journal of Ovarian Research 11: 1-13.
Lappöhn, R.E. & Burger, H.G. 1989. Bouma, J., Bangah, M., Krans, M. & de Bruijn, H.W. Inhibin as a marker for granulosa-cell tumors. The New England Journal of Medicine 21: 790-3.
La Marca A. & Volpe A. 2007. The Anti-Mullerian hormone and ovarian cancer. Human Reproduction Update 13: 265-273
McCue, P.M, Roser, J.F., Munro, C.J., Liu, I.K. & Lasley, B.L. 2006. Granulosa cell tumors of the equine ovary. Veterinary Clinics of North America: Equine Practice 22: 799-817.
Meimandi, K. & Yaghoobi, M.M. 2019. Effect of aqueous and ethanolic extracts of Sedum album L. on human stomach and breast carcinoma cell lines in vitro. Nova Biologica Reperta 6: 10-19
Mom, C.H., Engelen, M.J.A., Willemse, P.H.B., Gietema, J.A., ten Hoor, K.A., de Vries, E.G.E. & van der Zee, A.G.J. 2007. Granulosa cell tumors of the ovary: The clinical value of serum inhibin A and B levels in a large single center cohort. Gynecologic Oncology 105: 365-372
Monsefi, M., Nadi, A. & Alinejad, Z. 2017. The effects of Salvia officinalis L. on granulosa cells and in vitro maturation of oocytes in mice. International Journal of Reproductive BioMedicine 15: 649-660
Parisse, P., Rago, I., Ulloa Severino, L., Perissinotto, F., Ambrosetti, E., Paoletti, P., Ricci, M., Beltrami, A.P., Cesselli, D. & Casalis, L. 2017. Atomic force microscopy analysis of extracellular vesicles. European Biophysics Journal 46: 813-820.
Peak, T.C, Praharaj, P.P., Panigrahi, G.K., Doyle, M., Su, Y., Schlaepfer, I.R., Singh, R., Vander Griend, D.J., Alickson, J., Hemal, A., Atala, A. & Deep, G. 2018. Exosomes secreted by placental stem cells selectively inhibit growth of aggressive prostate cancer cells. Biochemical and Biophysical Research Communications 499: 1004-1010.
Phyoe-Battaglia, T., Bartels, C., Nulsen, J. & Grow, D.R. 2018. In vitro fertilization with granulosa cell tumor: a report of two cases. Journal of Assisted Reproduction and Genetics 35: 1919-1921.
Sharma, S., LeClaire, M. & Gimzewski, J.K. 2018. Ascent of atomic force microscopy as a nanoanalytical tool for exosomes and other extracellular vesicles. Nanotechnology 29: 1-20.
Sokolova, V., Ludwig, A.K., Hornung, S., Rotan, O., Horn, P.A., Epple, M. & Giebel, B. 2011. Characterisation of exosomes derived from human cells by nanoparticle tracking analysis and scanning electron microscopy. Colloids and Surfaces B: Biointerfaces 87: 146-50.
Stefanius, K., Servage, K., Santos, M.D.S., Gray, H.F., E Toomb, J., Chimalapati S., Kim, M.S., Malladi, V.S., Brekken, R. & Orth, K. 2019. Human pancreatic cancer cell exosomes, but not human normal cell exosomes, act as an initiator in cell transformation. eLife 8: 1-26
Van der Pol, E., Hoekstra, A.G., Sturk, A., Otto, C., van Leeuwen, T.G. & Nieuwland, R. 2010. Optical and non-optical methods for detection and characterization of microparticles and exosomes. Journal of Thrombosis and Haemostasis 8: 2596-607.
Vanderfiyden, B C., Telfer, E.E. & Eppig, J.J. 1992. Mouse oocytes promote proliferation of granulosa cells from preantral and antral follicles in vitro. Biology of Reproduction 46, 1196-1204.
Wu, Y., Deng, W. & Klinke, D.J. 2015. Exosomes: improved methods to characterize their morphology, RNA content, and surface protein biomarkers. Analyst 140: 6631-42.
Zhu, H., Yang, S.Y., Wang, J., Wang, L. & Han, S.Y. 2016. Evidence for miR-17-92 and miR-134 gene cluster regulation of ovarian cancer drug resistance. European Review for Medical and Pharmacological Sciences 20: 25-31.

  • تاریخ دریافت 18 خرداد 1405
  • تاریخ انتشار 18 خرداد 1405

صفحه اصلی

راهنمای نویسندگان

ارسال مقاله

داوران

تماس با ما