Advances in Molecular OncologyAdvances in Molecular OncologyУспехи молекулярной онкологии2313-805X2413-3787Publishing House ABV Press65410.17650/2313-805X-2024-11-1-113-123EXPERIMENTAL REPORTЭКСПЕРИМЕНТАЛЬНАЯ СТАТЬЯAnalysis of miRNAs miR-125a-5p, -27a-5p, -193a-5p, -135b-5p, -451a, -495-3p and -136-5p in parental ovarian cancer cells and secreted extracellular vesiclesАнализ микроРНК miR-125a-5p, -27a-5p, -193a-5p, -135b-5p, -451а, -495-3р и -136-5р в клетках рака яичника и секретируемых ими экстраклеточных везикулахhttps://orcid.org/0000-0002-4127-6973SkryabinG. O.СкрябинГ. О.
Russian Federation

24 Kashirskoye Shosse, Moscow 115522

115522 Москва, Каширское шоссе, 24

https://orcid.org/0000-0002-9364-562XBeliaevaA. A.БеляеваА. А.
Russian Federation

24 Kashirskoye Shosse, Moscow 115522; 1–12 Leninskie Gory, Moscow 119991

115522 Москва, Каширское шоссе, 24; 119234 Москва, Ленинские горы, 1, стр. 12

https://orcid.org/0000-0002-7628-8616EnikeevA. D.ЕникеевА. Д.
Russian Federation

24 Kashirskoye Shosse, Moscow 115522

115522 Москва, Каширское шоссе, 24

https://orcid.org/0000-0002-6355-7282BagrovD. V.БагровД. В.
Russian Federation

1–12 Leninskie Gory, Moscow 119991

119234 Москва, Ленинские горы, 1, стр. 12

https://orcid.org/0009-0005-5399-914XKeremetA. M.КереметА. М.
Russian Federation

1–12 Leninskie Gory, Moscow 119991

119234 Москва, Ленинские горы, 1, стр. 12

https://orcid.org/0000-0003-0766-163XKomelkovА. V.КомельковА. В.
Russian Federation

24 Kashirskoye Shosse, Moscow 115522

115522 Москва, Каширское шоссе, 24

https://orcid.org/0000-0002-4793-6063ElkinD. S.ЕлкинД. С.
Russian Federation

24 Kashirskoye Shosse, Moscow 115522

115522 Москва, Каширское шоссе, 24

https://orcid.org/0009-0009-1536-5778SylantievaD. M.СилантьеваД. М.
Russian Federation

1 Ostrovityanova St., Moscow 117997

117997 Москва, ул. Островитянова, 1

https://orcid.org/0000-0001-8837-7969TchevkinaE. M.ЧевкинаЕ. М.
Russian Federation

24 Kashirskoye Shosse, Moscow 115522

Елена Максимовна Чевкина

115522 Москва, Каширское шоссе, 24

tchevkina@mail.ruN.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of the RussiaФГБУ «Национальный медицинский исследовательский центр онкологии им. Н.Н. Блохина» Минздрава РоссииM.V. Lomonosov Moscow State UniversityФГБОУ ВО «Московский государственный университет им. М.В. Ломоносова»Pirogov Russian National Research Medical UniversityФГАОУ ВО «Российский национальный исследовательский медицинский университет им. Н.И. Пирогова» Минздрава России150120241111131230504202405042024Copyright ©; 2024, Skryabin G.O., Beliaeva A.A., Enikeev A.D., Bagrov D.V., Keremet A.M., Komelkov А.V., Elkin D.S., Sylantieva D.M., Tchevkina E.M.Copyright ©; 2024, Скрябин Г.О., Беляева А.А., Еникеев А.Д., Багров Д.В., Керемет А.М., Комельков А.В., Елкин Д.С., Силантьева Д.М., Чевкина Е.М.2024Skryabin G.O., Beliaeva A.A., Enikeev A.D., Bagrov D.V., Keremet A.M., Komelkov А.V., Elkin D.S., Sylantieva D.M., Tchevkina E.M.Скрябин Г.О., Беляева А.А., Еникеев А.Д., Багров Д.В., Керемет А.М., Комельков А.В., Елкин Д.С., Силантьева Д.М., Чевкина Е.М.https://creativecommons.org/licenses/by/4.0https://umo.abvpress.ru/jour/article/view/654

Introduction. The identification of markers for liquid diagnostics of ovarian cancer is one of the most urgent tasks of gynecologic oncology. Currently, extracellular vesicles (EVs) are of great interest as a source of oncomarkers, including miRNA markers. We have previously shown that the levels of miR-125a-5p, -27a-5p, -193a-5p and 135b-5p are significantly elevated and miR-451a, -495-3p and -136-5p are significantly decreased in the EVs from uterine aspirates of ovarian cancer patients.

Aim. Analysis of miR-125a-5p, -27a-5p, -193a-5p, 135b-5p, 451a, 495-3p and -136-5p levels in ovarian cancer cell cultures and secreted EVs.

Material and methods. Cultivation of ovarian cancer cell lines: OVCAR-3, OVCAR-4, OVCAR-8 and SKOV3; EVs isolation from conditioned medium by ultracentrifugation; EVs validation by nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), western blot analysis of exosomal markers; isolation of miRNAs from cells and EVs; analysis of miRNAs by Stem-Loop – reverse transcription-quantitative polymerase chain reaction.

Results. In all cell lines studied, the expression of miR-125a-5p, -27a-5p, -193a-5p and -135b-5p significantly exceeds the expression of -451a, -495-3p and -136-5p. All ovarian cancer cell lines are featured by a “cells >EVs” ratio for highly expressed miRNAs and “EVs >cells” ratio for poorly expressed miRNAs.

Conclusion. The results of the study support the relation between the differential expression of studied miRNAs and the pathogenesis of ovarian cancer and confirm the high diagnostic potential of these molecules.

Введение. Поиск маркеров для жидкостной диагностики рака яичника (РЯ) является одной из наиболее актуальных задач онкогинекологии. В настоящее время большой интерес в качестве источника онкомаркеров, в том числе микроРНК, вызывают экстраклеточные везикулы (ЭКВ). Ранее мы показали, что уровень miR-125a-5p, -27a-5p, -193a-5p и -135b-5p достоверно повышен, а miR-451а, -495-3р и -136-5р значимо снижен в ЭКВ маточных аспиратов больных РЯ.

Цель исследования – анализ уровней miR-125a-5p, -27a-5p, -193a-5p, -135b-5p, -451а, -495-3р и -136-5р в клеточных линиях РЯ и секретируемых ими ЭКВ.

Материалы и методы. Проведены культивирование клеточных линий РЯ (OVCAR-3, OVCAR-4, OVCAR-8 и SKOV3), выделение ЭКВ из кондиционированной среды методом ультрацентрифугирования, валидация ЭКВ с помощью анализа траекторий наночастиц (NTA), трансмиссионной электронной микроскопии и вестерн-блот-анализа экзосомальных маркеров. Также выполнены выделение микроРНК из клеток и ЭКВ, анализ микроРНК методом полимеразной цепной реакции с обратной транскрипцией в реальном времени в модификации Stem-loop.

Результаты. В клетках исследуемых линий РЯ экспрессия молекул miR-125a-5p, -27a-5p, -193a-5p и -135b-5p значительно превышала экспрессию miR-451а, -495-3р и -136-5р. Все линии клеток РЯ характеризуются соотношением «клетки >ЭКВ» для высоко экспрессируемых микроРНК и «ЭКВ >клетки» для низко экспрессируемых микроРНК.

Заключение. Результаты исследования свидетельствуют о связи дифференциальной экспрессии исследуемых микроРНК с патогенезом РЯ и подтверждают высокий диагностический потенциал данных молекул.

ovarian cancerextracellular vesiclesexosomesmiRNAmiR-125a-5pmiR-27a-5pmiR-193a-5pmiR-135b5pmiR-451amiR-495-3pmiR-136-5ррак яичникаэкстраклеточные везикулыэкзосомымикроРНКmiR-125a-5pmiR-27a-5pmiR-193a5pmiR-135b-5pmiR-451аmiR-495-3рmiR-136-5рThis research was funded by the Russian Scientific Foundation (grant No. 22-15-00373)Работа выполнена при финансовой поддержке Российского научного фонда (грант № 22-15-00373)1. Raposo G., Stahl P.D. Extracellular vesicles – on the cusp of a new language in the biological sciences. Extracell Vesicles Circ Nucleic Acids 2023;4(2):240–54. DOI: 10.20517/evcna.2023.18Raposo G., Stahl P.D. Extracellular vesicles – on the cusp of a new language in the biological sciences. Extracell Vesicles Circ Nucleic Acids 2023;4(2):240–54. DOI: 10.20517/evcna.2023.182. Kalluri R., LeBleu V.S. The biology, function, and biomedical applications of exosomes. Science 2020;367(6478):eaau6977. DOI: 10.1126/science.aau6977Kalluri R., LeBleu V.S. The biology, function, and biomedical applications of exosomes. Science 2020;367(6478):eaau6977. DOI: 10.1126/science.aau69773. Liu Y.-J., Wang C. A review of the regulatory mechanisms of extracellular vesicles-mediated intercellular communication. Cell Commun Signal 2023;21(1):77. DOI: 10.1186/s12964-023-01103-6Liu Y.-J., Wang C. A review of the regulatory mechanisms of extracellular vesicles-mediated intercellular communication. Cell Commun Signal 2023;21(1):77. DOI: 10.1186/s12964-023-01103-64. Xu R., Rai A., Chen M. et al. Extracellular vesicles in cancer – implications for future improvements in cancer care. Nat Rev Clin Oncol 2018;15(10):617–38. DOI: 10.1038/s41571-018-0036-9Xu R., Rai A., Chen M. et al. Extracellular vesicles in cancer – implications for future improvements in cancer care. Nat Rev Clin Oncol 2018;15(10):617–38. DOI: 10.1038/s41571-018-0036-95. Logozzi M., Mizzoni D., Di Raimo R., Fais S. Exosomes: a source for new and old biomarkers in cancer. Cancers 2020;12(9):2566. DOI: 10.3390/cancers12092566Logozzi M., Mizzoni D., Di Raimo R., Fais S. Exosomes: a source for new and old biomarkers in cancer. Cancers 2020;12(9):2566. DOI: 10.3390/cancers120925666. Staicu C.E., Predescu D.V., Rusu C.M. et al. Role of microRNAs as clinical cancer biomarkers for ovarian cancer: a short overview. Cells 2020;9(1):169. DOI: 10.3390/cells9010169Staicu C.E., Predescu D.V., Rusu C.M. et al. Role of microRNAs as clinical cancer biomarkers for ovarian cancer: a short overview. Cells 2020;9(1):169. DOI: 10.3390/cells90101697. Meng X., Müller V., Milde-Langosch K. et al. Diagnostic and prognostic relevance of circulating exosomal miR-373, miR-200a, miR-200b and miR-200c in patients with epithelial ovarian cancer. Oncotarget 2016;7(13):16923–35. DOI: 10.18632/oncotarget.7850Meng X., Müller V., Milde-Langosch K. et al. Diagnostic and prognostic relevance of circulating exosomal miR-373, miR-200a, miR-200b and miR-200c in patients with epithelial ovarian cancer. Oncotarget 2016;7(13):16923–35. DOI: 10.18632/oncotarget.78508. Pan C., Stevic I., Müller V. et al. Exosomal microRNAs as tumor markers in epithelial ovarian cancer. Mol Oncol 2018;12(11):1935–48. DOI: 10.1002/1878-0261.12371Pan C., Stevic I., Müller V. et al. Exosomal microRNAs as tumor markers in epithelial ovarian cancer. Mol Oncol 2018;12(11):1935–48. DOI: 10.1002/1878-0261.123719. Théry C., Witwer K.W., Aikawa E. et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles 2018;7(1):1535750. DOI: 10.1080/20013078.2018.1535750Théry C., Witwer K.W., Aikawa E. et al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles 2018;7(1):1535750. DOI: 10.1080/20013078.2018.153575010. Salmond N., Williams K.C. Isolation and characterization of extracellular vesicles for clinical applications in cancer – time for standardization? Nanoscale Adv 2021;3(7):1830–52. DOI: 10.1039/d0na00676aSalmond N., Williams K.C. Isolation and characterization of extracellular vesicles for clinical applications in cancer – time for standardization? Nanoscale Adv 2021;3(7):1830–52. DOI: 10.1039/d0na00676a11. Skryabin G.O., Komelkov A.V., Zhordania K.I. et al. Extracellular vesicles from uterine aspirates represent a promising source for screening markers of gynecologic cancers. Cells 2022;11(7):1064. DOI: 10.3390/cells11071064Skryabin G.O., Komelkov A.V., Zhordania K.I. et al. Extracellular vesicles from uterine aspirates represent a promising source for screening markers of gynecologic cancers. Cells 2022;11(7):1064. DOI: 10.3390/cells1107106412. Kramer F. Stem-Loop RT-qPCR for miRNAs. Curr Protoc Mol Biol 2011;Chapter 15:Unit15.10. DOI: 10.1002/0471142727.mb1510s95Kramer F. Stem-Loop RT-qPCR for miRNAs. Curr Protoc Mol Biol 2011;Chapter 15:Unit15.10. DOI: 10.1002/0471142727.mb1510s9513. Skryabin G.O., Komelkov A.V., Galetsky S.A. et al. Stomatin is highly expressed in exosomes of different origin and is a promising candi-date as an exosomal marker. J Cell Biochem 2021;122(1):100–15. DOI: 10.1002/jcb.29834Skryabin G.O., Komelkov A.V., Galetsky S.A. et al. Stomatin is highly expressed in exosomes of different origin and is a promising candi-date as an exosomal marker. J Cell Biochem 2021;122(1):100–15. DOI: 10.1002/jcb.2983414. Burdiel M., Jiménez J., Rodríguez-Antolín C. et al. MiR-151a: a robust endogenous control for normalizing small extracellular vesicle cargo in human cancer. Biomark Res 2023;11(1): 94. DOI: 10.1186/s40364-023-00526-0Burdiel M., Jiménez J., Rodríguez-Antolín C. et al. MiR-151a: a robust endogenous control for normalizing small extracellular vesicle cargo in human cancer. Biomark Res 2023;11(1): 94. DOI: 10.1186/s40364-023-00526-015. Xie F., Wang J., Zhang B. RefFinder: a web-based tool for comprehensively analyzing and identifying reference genes. Funct Integr Genomics 2023;23(2):125. DOI: 10.1007/s10142-023-01055-7Xie F., Wang J., Zhang B. RefFinder: a web-based tool for comprehensively analyzing and identifying reference genes. Funct Integr Genomics 2023;23(2):125. DOI: 10.1007/s10142-023-01055-716. Wang X., Huang J., Chen W. et al. The updated role of exosomal proteins in the diagnosis, prognosis, and treatment of cancer. Exp Mol Med 2022;54(9):1390–400. DOI: 10.1038/s12276-022-00855-4Wang X., Huang J., Chen W. et al. The updated role of exosomal proteins in the diagnosis, prognosis, and treatment of cancer. Exp Mol Med 2022;54(9):1390–400. DOI: 10.1038/s12276-022-00855-417. Zhang J., Li S., Li L. et al. Exosome and exosomal microRNA: trafficking, sorting, and function. Genomics Proteomics Bioinformatics 2015;13(1):17–24. DOI: 10.1016/j.gpb.2015.02.001Zhang J., Li S., Li L. et al. Exosome and exosomal microRNA: trafficking, sorting, and function. Genomics Proteomics Bioinformatics 2015;13(1):17–24. DOI: 10.1016/j.gpb.2015.02.00118. Liu Q.-W., He Y., Xu W.W. Molecular functions and therapeutic applications of exosomal noncoding RNAs in cancer. Exp Mol Med 2022;54(3):216–25. DOI: 10.1038/s12276-022-00744-wLiu Q.-W., He Y., Xu W.W. Molecular functions and therapeutic applications of exosomal noncoding RNAs in cancer. Exp Mol Med 2022;54(3):216–25. DOI: 10.1038/s12276-022-00744-w19. Skryabin G.O., Vinokurova S.V., Elkina N.V. et al. Comparison of methods for microRNA isolation from extracellular vesicles obtained from ascitic fluids. Biochemistry 2022;87(11):1354–66. DOI: 10.1134/S0006297922110141Skryabin G.O., Vinokurova S.V., Elkina N.V. et al. Comparison of methods for microRNA isolation from extracellular vesicles obtained from ascitic fluids. Biochemistry 2022;87(11):1354–66. DOI: 10.1134/S000629792211014120. Koutsaki M., Libra M., Spandidos D.A., Zaravinos A. The miR-200 family in ovarian cancer. Oncotarget 2017;8(39):66629–40. DOI: 10.18632/oncotarget.18343Koutsaki M., Libra M., Spandidos D.A., Zaravinos A. The miR-200 family in ovarian cancer. Oncotarget 2017;8(39):66629–40. DOI: 10.18632/oncotarget.1834321. Liu X., Li J., Qin F., Dai S. miR-152 as a tumor suppressor microRNA: target recognition and regulation in cancer. Oncol Lett 2016;11(6):3911–6. DOI: 10.3892/ol.2016.4509Liu X., Li J., Qin F., Dai S. miR-152 as a tumor suppressor microRNA: target recognition and regulation in cancer. Oncol Lett 2016;11(6):3911–6. DOI: 10.3892/ol.2016.450922. Xuan J., Liu Y., Zeng X., Wang H. Sequence requirements for miR-424-5p regulating and function in cancers Int J Mol Sci 2022;23(7):4037. DOI: 10.3390/ijms23074037Xuan J., Liu Y., Zeng X., Wang H. Sequence requirements for miR-424-5p regulating and function in cancers Int J Mol Sci 2022;23(7):4037. DOI: 10.3390/ijms2307403723. Timofeeva A.V., Fedorov I.S., Asaturova A.V. et al. Blood plasma small non-coding RNAs as diagnostic molecules for the progesterone-receptor-negative phenotype of serous ovarian tumors. Int J Mol Sci 2023;24(15):12214. DOI: 10.3390/ijms241512214Timofeeva A.V., Fedorov I.S., Asaturova A.V. et al. Blood plasma small non-coding RNAs as diagnostic molecules for the progesterone-receptor-negative phenotype of serous ovarian tumors. Int J Mol Sci 2023;24(15):12214. DOI: 10.3390/ijms24151221424. Gadducci A., Sergiampietri C., Lanfredini N., Guiggi I. MicroRNAs and ovarian cancer: the state of art and perspectives of clinical research. Gynecol Endocrinol 2014;30(4):266–71. DOI: 10.3109/09513590.2013.871525Gadducci A., Sergiampietri C., Lanfredini N., Guiggi I. MicroRNAs and ovarian cancer: the state of art and perspectives of clinical research. Gynecol Endocrinol 2014;30(4):266–71. DOI: 10.3109/09513590.2013.87152525. Jiang Y., Shi Y., Lyu T. et al. Identification and functional validation of differentially expressed microRNAs in ascites-derived ovarian cancer cells compared with primary tumour tissue. Cancer Manag Res 2021;13:6585–97. DOI: 10.2147/CMAR.S320834Jiang Y., Shi Y., Lyu T. et al. Identification and functional validation of differentially expressed microRNAs in ascites-derived ovarian cancer cells compared with primary tumour tissue. Cancer Manag Res 2021;13:6585–97. DOI: 10.2147/CMAR.S32083426. Wang J., Zhang R., Zhang B. et al. MiR-135b improves proliferation and regulates chemotherapy resistance in ovarian cancer. J Mol Histol 2022;53(4):699–712. DOI: 10.1007/s10735-022-10080-yWang J., Zhang R., Zhang B. et al. MiR-135b improves proliferation and regulates chemotherapy resistance in ovarian cancer. J Mol Histol 2022;53(4):699–712. DOI: 10.1007/s10735-022-10080-y27. Chen H., Mao M., Jiang J.D. et al. Circular RNA CDR1as acts as a sponge of miR-135b-5p to suppress ovarian cancer progression. OncoTargets Ther 2019;12:3869–79. DOI: 10.2147/OTT.S207938Chen H., Mao M., Jiang J.D. et al. Circular RNA CDR1as acts as a sponge of miR-135b-5p to suppress ovarian cancer progression. OncoTargets Ther 2019;12:3869–79. DOI: 10.2147/OTT.S20793828. Yu S., Yu M., Chen J. et al. Circ_0000471 suppresses the progression of ovarian cancer through mediating mir-135b-5p/dusp5 axis. Am J Reprod Immunol 2023;89(4):e13651. DOI: 10.1111/aji.13651Yu S., Yu M., Chen J. et al. Circ_0000471 suppresses the progression of ovarian cancer through mediating mir-135b-5p/dusp5 axis. Am J Reprod Immunol 2023;89(4):e13651. DOI: 10.1111/aji.1365129. Cao Y., Shen T., Zhang C. et al. MiR-125a-5p inhibits EMT of ovarian cancer cells by regulating TAZ/EGFR signaling pathway. Eur Rev Med Pharmacol Sci 2019;23(19):8249–56. DOI: 10.26355/eurrev_201910_19134Cao Y., Shen T., Zhang C. et al. MiR-125a-5p inhibits EMT of ovarian cancer cells by regulating TAZ/EGFR signaling pathway. Eur Rev Med Pharmacol Sci 2019;23(19):8249–56. DOI: 10.26355/eurrev_201910_1913430. Lee M., Kim E.J., Jeon M.J. MicroRNAs 125a and 125b inhibit ovarian cancer cells through post-transcriptional inactivation of EIF4EBP1. Oncotarget 2015;7(8):8726–42. DOI: 10.18632/oncotarget.6474Lee M., Kim E.J., Jeon M.J. MicroRNAs 125a and 125b inhibit ovarian cancer cells through post-transcriptional inactivation of EIF4EBP1. Oncotarget 2015;7(8):8726–42. DOI: 10.18632/oncotarget.647431. Yang J., Li G., Zhang K. MiR-125a regulates ovarian cancer proliferation and invasion by repressing GALNT14 expression. Biomed Pharmacother 2016;80:381–7. DOI: 10.1016/j.biopha.2015.12.027Yang J., Li G., Zhang K. MiR-125a regulates ovarian cancer proliferation and invasion by repressing GALNT14 expression. Biomed Pharmacother 2016;80:381–7. DOI: 10.1016/j.biopha.2015.12.02732. Wang Y., Li N., Zhao J., Dai C. MiR-193a-5p serves as an inhibitor in ovarian cancer cells through RAB11A. Reprod Toxicol 2022;110:105–12. DOI: 10.1016/j.reprotox.2022.04.003Wang Y., Li N., Zhao J., Dai C. MiR-193a-5p serves as an inhibitor in ovarian cancer cells through RAB11A. Reprod Toxicol 2022;110:105–12. DOI: 10.1016/j.reprotox.2022.04.00333. Zhang S., Liu J., He J., Yi N. MicroRNA-193a-5p exerts a tumor suppressive role in epithelial ovarian cancer by modulating RBBP6. Mol Med Rep 2021;24(2):582. DOI: 10.3892/mmr.2021.12221Zhang S., Liu J., He J., Yi N. MicroRNA-193a-5p exerts a tumor suppressive role in epithelial ovarian cancer by modulating RBBP6. Mol Med Rep 2021;24(2):582. DOI: 10.3892/mmr.2021.1222134. Khordadmehr M., Shahbazi R., Sadreddini S., Baradaran B. miR-193: a new weapon against cancer. J Cell Physiol 2019;234(10): 6861–72. DOI: 10.1002/jcp.28368Khordadmehr M., Shahbazi R., Sadreddini S., Baradaran B. miR-193: a new weapon against cancer. J Cell Physiol 2019;234(10): 6861–72. DOI: 10.1002/jcp.2836835. Eitan R., Kushnir M., Lithwick-Yanai G. et al. Tumor microRNA expression patterns associated with resistance to platinum based chemotherapy and survival in ovarian cancer patients. Gynecol Oncol 2009;114(2):253–9. DOI: 10.1016/j.ygyno.2009.04.024Eitan R., Kushnir M., Lithwick-Yanai G. et al. Tumor microRNA expression patterns associated with resistance to platinum based chemotherapy and survival in ovarian cancer patients. Gynecol Oncol 2009;114(2):253–9. DOI: 10.1016/j.ygyno.2009.04.02436. Wambecke A., Ahmad M., Morice P.M. et al. The lncRNA ‘UCA1’ modulates the response to chemotherapy of ovarian cancer through direct binding to miR-27a-5p and control of UBE2N levels. Mol Oncol 2021;15(12):3659–78. DOI: 10.1002/1878-0261.13045Wambecke A., Ahmad M., Morice P.M. et al. The lncRNA ‘UCA1’ modulates the response to chemotherapy of ovarian cancer through direct binding to miR-27a-5p and control of UBE2N levels. Mol Oncol 2021;15(12):3659–78. DOI: 10.1002/1878-0261.1304537. Che X., Jian F., Chen C. et al. PCOS serum-derived exosomal miR-27a-5p stimulates endometrial cancer cells migration and invasion. J Mol Endocrinol 2020;64(1):1–12. DOI: 10.1530/JME-19-0159Che X., Jian F., Chen C. et al. PCOS serum-derived exosomal miR-27a-5p stimulates endometrial cancer cells migration and invasion. J Mol Endocrinol 2020;64(1):1–12. DOI: 10.1530/JME-19-015938. Regis S., Caliendo F., Dondero A. et al. TGF-β1 downregulates the expression of CX3CR1 by inducing miR-27a-5p in primary human NK cells. Front Immunol 2017;8. Available at: https://www.frontiersin.org/articles/10.3389/fimmu.2017.00868Regis S., Caliendo F., Dondero A. et al. TGF-β1 downregulates the expression of CX3CR1 by inducing miR-27a-5p in primary human NK cells. Front Immunol 2017;8. Available at: https://www.frontiersin.org/articles/10.3389/fimmu.2017.0086839. Huldani H., Malviya J., Rodrigues P. et al. miR-495–3p as a promising tumor suppressor in human cancers. Pathol Res Pract 2023;248:154610. DOI: 10.1016/j.prp.2023.154610Huldani H., Malviya J., Rodrigues P. et al. miR-495–3p as a promising tumor suppressor in human cancers. Pathol Res Pract 2023;248:154610. DOI: 10.1016/j.prp.2023.15461040. Chen H., Wang X., Bai J., He A. Expression, regulation and function of miR-495 in healthy and tumor tissues. Oncol Lett 2017;13(4):2021–6. DOI: 10.3892/ol.2017.5727Chen H., Wang X., Bai J., He A. Expression, regulation and function of miR-495 in healthy and tumor tissues. Oncol Lett 2017;13(4):2021–6. DOI: 10.3892/ol.2017.572741. Liu S., Xi X. LINC01133 contribute to epithelial ovarian cancer metastasis by regulating miR-495-3p/TPD52 axis. Biochem Biophys Res Commun 2020;533(4):1088–94. DOI: 10.1016/j.bbrc.2020.09.074Liu S., Xi X. LINC01133 contribute to epithelial ovarian cancer metastasis by regulating miR-495-3p/TPD52 axis. Biochem Biophys Res Commun 2020;533(4):1088–94. DOI: 10.1016/j.bbrc.2020.09.07442. Zhu J., Luo J.E., Chen Y., Wu Q. Circ_0061140 knockdown inhibits tumorigenesis and improves PTX sensitivity by regulating miR-136/ CBX2 axis in ovarian cancer. J Ovarian Res 2021;14(10):136. DOI: 10.1186/s13048-021-00888-9Zhu J., Luo J.E., Chen Y., Wu Q. Circ_0061140 knockdown inhibits tumorigenesis and improves PTX sensitivity by regulating miR-136/ CBX2 axis in ovarian cancer. J Ovarian Res 2021;14(10):136. DOI: 10.1186/s13048-021-00888-943. Zhao H., Liu S., Wang G. et al. Expression of miR-136 is associated with the primary cisplatin resistance of human epithelial ovarian cancer. Oncol Rep 2015;33(2):591–8. DOI: 10.3892/or.2014.3640Zhao H., Liu S., Wang G. et al. Expression of miR-136 is associated with the primary cisplatin resistance of human epithelial ovarian cancer. Oncol Rep 2015;33(2):591–8. DOI: 10.3892/or.2014.364044. Ling S., Ruiqin M., Guohong Z., Ying W. Expression and prognostic significance of microRNA-451 in human epithelial ovarian cancer. Eur J Gynaecol Oncol 2015;36(4):463–8.Ling S., Ruiqin M., Guohong Z., Ying W. Expression and prognostic significance of microRNA-451 in human epithelial ovarian cancer. Eur J Gynaecol Oncol 2015;36(4):463–8.45. Zhu H., Wu H., Liu X. et al. Role of microRNA miR-27a and miR451 in the regulation of MDR1/P-glycoprotein expression in human cancer cells. Biochem Pharmacol 2008;76(5):582–8. DOI: 10.1016/j.bcp.2008.06.007Zhu H., Wu H., Liu X. et al. Role of microRNA miR-27a and miR451 in the regulation of MDR1/P-glycoprotein expression in human cancer cells. Biochem Pharmacol 2008;76(5):582–8. DOI: 10.1016/j.bcp.2008.06.00746. Bagnoli M., Canevari S., Califano D. et al. Development and validation of a microRNA-based signature (MiROvaR) to predict early relapse or progression of epithelial ovarian cancer: a cohort study. Lancet Oncol 2016;17(8):1137–46. DOI: 10.1016/S1470-2045(16)30108-5Bagnoli M., Canevari S., Califano D. et al. Development and validation of a microRNA-based signature (MiROvaR) to predict early relapse or progression of epithelial ovarian cancer: a cohort study. Lancet Oncol 2016;17(8):1137–46. DOI: 10.1016/S1470-2045(16)30108-547. De Cecco L., Bagnoli M., Chiodini P. et al. Prognostic evidence of the miRNA-based ovarian cancer signature MiROvaR in independent datasets. Cancers 2021;13(7):1544. DOI: 10.3390/cancers13071544De Cecco L., Bagnoli M., Chiodini P. et al. Prognostic evidence of the miRNA-based ovarian cancer signature MiROvaR in independent datasets. Cancers 2021;13(7):1544. DOI: 10.3390/cancers1307154448. Pucci M., Reclusa Asiáin P., Duréndez Sáez E. et al. Extracellular vesicles as miRNA nano-shuttles: dual role in tumor progression. Target Oncol 2018;13(2):175–87. DOI: 10.1007/s11523-018-0551-8Pucci M., Reclusa Asiáin P., Duréndez Sáez E. et al. Extracellular vesicles as miRNA nano-shuttles: dual role in tumor progression. Target Oncol 2018;13(2):175–87. DOI: 10.1007/s11523-018-0551-849. Guduric-Fuchs J., O’Connor A., Camp B. et al. Selective extracellular vesicle-mediated export of an overlapping set of microRNAs from multiple cell types. BMC Genomics 2012;13:357. DOI: 10.1186/1471-2164-13-357Guduric-Fuchs J., O’Connor A., Camp B. et al. Selective extracellular vesicle-mediated export of an overlapping set of microRNAs from multiple cell types. BMC Genomics 2012;13:357. DOI: 10.1186/1471-2164-13-35750. Ohshima K., Inoue K., Fujiwara A. et al. Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. PLoS One 2010;5(10): DOI: 10.1371/journal.pone.0013247Ohshima K., Inoue K., Fujiwara A. et al. Let-7 microRNA family is selectively secreted into the extracellular environment via exosomes in a metastatic gastric cancer cell line. PLoS One 2010;5(10): DOI: 10.1371/journal.pone.001324751. Bordanaba-Florit G., Madarieta I., Olalde B. et al. 3D cell cultures as prospective models to study extracellular vesicles in cancer. Cancers 2021;13(2):307. DOI: 10.3390/cancers13020307Bordanaba-Florit G., Madarieta I., Olalde B. et al. 3D cell cultures as prospective models to study extracellular vesicles in cancer. Cancers 2021;13(2):307. DOI: 10.3390/cancers1302030752. Kusuma G.D., Li A., Zhu D. et al. Effect of 2D and 3D culture microenvironments on mesenchymal stem cell-derived extracellular vesicles potencies. Front Cell Dev Biol 2022;10:819726. DOI: 10.3389/fcell.2022.819726Kusuma G.D., Li A., Zhu D. et al. Effect of 2D and 3D culture microenvironments on mesenchymal stem cell-derived extracellular vesicles potencies. Front Cell Dev Biol 2022;10:819726. DOI: 10.3389/fcell.2022.81972653. Rocha S., Carvalho J., Oliveira P. et al. 3D cellular architecture affects microRNA and protein cargo of extracellular vesicles. Adv Sci Weinh Baden-Wurtt Ger 2019;6(4):1800948. DOI: 10.1002/advs.201800948Rocha S., Carvalho J., Oliveira P. et al. 3D cellular architecture affects microRNA and protein cargo of extracellular vesicles. Adv Sci Weinh Baden-Wurtt Ger 2019;6(4):1800948. DOI: 10.1002/advs.20180094854. Thippabhotla S., Zhong C., He M. 3D cell culture stimulates the secretion of in vivo like extracellular vesicles. Sci Rep 2019;9(1):13012. DOI: 10.1038/s41598-019-49671-3Thippabhotla S., Zhong C., He M. 3D cell culture stimulates the secretion of in vivo like extracellular vesicles. Sci Rep 2019;9(1):13012. DOI: 10.1038/s41598-019-49671-3