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Ghorbani, M., Farimani, M. S., Khodadadi, I., Mohagheghi, S., Amiri, I., & Tayebinia, H. (2024). The regulatory roles of Smad2/3 protein and SMURF2 gene expression in granulosa cells of germinal vesicle and metaphase II oocytes in polycystic ovarian syndrome: A case-control study. International Journal of Reproductive BioMedicine (IJRM), 22(6), 441–450. https://doi.org/10.18502/ijrm.v22i6.16794
https://doi.org/10.18502/ijrm.v22i6.16794
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Marzieh Ghorbani
Marzieh Ghorbani
Department of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
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Marzieh Sanoee Farimani
Marzieh Sanoee Farimani
Department of Obstetrics and Gynecology, Medicine School, Hamadan University of Medical Sciences, Hamadan, Iran.
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Iraj Khodadadi
Iraj Khodadadi
Department of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
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Sina Mohagheghi
Sina Mohagheghi
Department of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
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Iraj Amiri
Iraj Amiri
Fertility and Infertility Research Centre, Hamadan University of Medical Sciences, Hamadan, Iran.
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Heidar Tayebinia
Heidar Tayebinia
Department of Clinical Biochemistry, School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
Published Date
- Jul 31, 2024
The regulatory roles of Smad2/3 protein and SMURF2 gene expression in granulosa cells of germinal vesicle and metaphase II oocytes in polycystic ovarian syndrome: A case-control study
Abstract
Background: The impaired functions of granulosa cells (GCs) in the delayed development and immaturity of oocytes have been reported in polycystic ovary syndrome (PCOs). Even with ovarian stimulation, a large number of oocytes in these patients are still in the stage germinal vesicle (GV).
Objective: The levels of Smad2/3, phosphorylated Smad2/3 (P-Smad2/3), the expression of SARA, Smad4, and SMURF2 genes in the GCs surrounding metaphase II (MII) or GV oocytes in PCOs women were investigated.
Materials and Methods: GCs of MII and GV oocytes were isolated from 38 women with PCOs and the expression levels of SARA, Smad4, and SMURF2 in surrounding GCs of MII and GV oocytes were determined using reverse-transcription polymerase chain reaction. Also, Smad2/3 and P-Smad2/3 proteins were determined using western blotting.
Results: The expression level of SMURF2 was significantly higher in GCs surrounding GV oocytes compared with that of GCs encompassing MII oocytes (p < 0.001). At the same time, no significant differences were observed in SARA and Smad4 expression levels in GCs surrounding GV and MII oocytes. A lower level of P-Smad2/3 was also found in GCs GV oocytes compared with GCs of MII oocytes (p < 0.001).
Conclusion: It seems that P-Smad2/3 plays a role in oocyte development, and the downregulation of this protein is associated with a defect in the maturation of GV oocytes. On the other hand, the upregulation of the SMURF2 gene also affects the growth process of GCs and the maturation of GV oocytes.
Key words: Polycystic ovary, Oocytes, Granulosa cell.
References
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[2] Zeng X, Xie Y-J, Liu Y-T, Long S-l, Mo Z-C. Polycystic ovarian syndrome: Correlation between hyperandrogenism, insulin resistance and obesity. Clin Chim Acta 2020; 502: 214–221.
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[5] Pedroso DC, Santana VP, Donaires FS, Picinato MC, Giorgenon RC, Santana BA, et al. Telomere length and telomerase activity in immature oocytes and cumulus cells of women with polycystic ovary syndrome. Reprod Sci 2020; 27: 1293–1303.
[6] de Moraes FP, Missio D, Lazzari J, Rovani MT, Ferreira R, Gonçalves PBD, et al. Local regulation of antral follicle development and ovulation in monovulatory species. Anim Reprod 2023; 19: e20220099.
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[10] Wang Z, Wei H, Wu Z, Zhang X, Sun Y, Gao L, et al. The oocyte cumulus complex regulates mouse sperm migration in the oviduct. Commun Biol 2022; 5: 1327.
[11] Rozés-Salvador V, Siri SO, Musri MM, Conde C. New player in endosomal trafficking: Differential roles of Smad anchor for receptor activation (SARA) protein. Mol Cell Biol 2018; 38: e00446.
[12] Tan Y, Chen Y, Du M, Peng Z, Xie P. USF2 inhibits the transcriptional activity of Smurf1 and SMURF2 to promote breast cancer tumorigenesis. Cell Signal 2019; 53: 49–58.
[13] Rotterdam ESHRE/ASRM-Sponsored PCOS consensus workshop group. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome (PCOS). Hum Reprod 2004; 19: 41–47.
[14] Gilchrist RB, Smitz J. Oocyte in vitro maturation: Physiological basis and application to clinical practice. Fertil Steril 2023; 119: 524–539.
[15] Li Y, Xiong G, Tan J, Wang S, Wu Q, Wan L, et al. Aberrant activation of the Hedgehog signaling pathway in granulosa cells from patients with polycystic ovary syndrome. Bioengineered 2021; 12: 12123–12134.
[16] Huang R-H, Zhou W-H. Granulosa cell biomarkers to predict oocyte and embryo quality in assisted reproductive technology. Reprod Dev Med 2021; 5: 30–37.
[17] Qin N, Fan X-C, Xu X-X, Tyasi TL, Li S-J, Zhang Y-Y, et al. Cooperative effects of FOXL2 with the members of TGF-?? superfamily on FSH receptor mRNA expression and granulosa cell proliferation from hen prehierarchical follicles. PLoS One 2015; 10: e0141062.
[18] Bai L, Chang HM, Zhang L, Zhu YM, Leung PC. BMP2 increases the production of BDNF through the upregulation of proBDNF and furin expression in human granulosa-lutein cells. FASEB J 2020; 34: 16129–16143.
[19] Tzavlaki K, Moustakas A. TGF-?? Signaling. Biomolecules 2020; 10: 487.
[20] Clark KL, George JW, Przygrodzka E, Plewes MR, Hua G, Wang C, et al. Hippo signaling in the ovary: Emerging roles in development, fertility, and disease. Endocr Rev 2022; 43: 1074–1096.
[21] Gong Z, Yang J, Bai S, Wei S. MicroRNAs regulate granulosa cells apoptosis and follicular development- A review. Asian-Australas J Anim Sci 2020; 33: 1714–1724.
[22] Shi XY, Guan ZQ, Yu JN, Liu HL. Follicle stimulating hormone inhibits the expression of p53 up-regulated modulator of apoptosis induced by reactive oxygen species through PI3K/AKT in mouse granulosa cells. Physiol Res 2020; 69: 687–694.
[23] Shen M, Liu Z, Li B, Teng Y, Zhang J, Tang Y, et al. Involvement of FoxO1 in the effects of follicle-stimulating hormone on inhibition of apoptosis in mouse granulosa cells. Cell Death Dis 2014; 5: e1475.
[24] Bhardwaj JK, Paliwal A, Saraf P, Sachdeva SN. Role of autophagy in follicular development and maintenance of primordial follicular pool in the ovary. J Cell Physiol 2022; 237: 1157–1170.
[25] de Ceuninck van Capelle C, Spit M, Ten Dijke P. Current perspectives on inhibitory Smad7 in health and disease. Crit Rev Biochem Mol 2020; 55: 691–715.
[26] Baddela VS, Michaelis M, Sharma A, Plinski C, Viergutz T, Vanselow J. Estradiol production of granulosa cells is unaffected by the physiological mix of nonesterified fatty acids in follicular fluid. Biol Chem 2022; 298: 691–715.
[27] Du X, Li Q, Yang L, Liu L, Cao Q, Li Q. Smad4 activates Wnt signaling pathway to inhibit granulosa cell apoptosis. Cell Death Dis 2020; 11: 373.
[28] Liu L, Li Q, Yang L, Li Q, Du X. Smad4 feedback activates the canonical TGF-?? family signaling pathways. Int J Mol Sci 2021; 22: 10024.
[29] Piotrowska H, Kempisty B, Sosinska P, Ciesiolka S, Bukowska D, Antosik P, et al. The role of TGF superfamily gene expression in the regulation of folliculogenesis and oogenesis in mammals: a review. Vet Med 2013; 58: 505– 515.
[30] Chandhoke AS, Karve K, Dadakhujaev S, Netherton S, Deng L, Bonni S. The ubiquitin ligase SMURF2 suppresses TGF??-induced epithelial- mesenchymal transition in a sumoylation-regulated manner. Cell Death Differ 2016; 23: 876–888.
[2] Zeng X, Xie Y-J, Liu Y-T, Long S-l, Mo Z-C. Polycystic ovarian syndrome: Correlation between hyperandrogenism, insulin resistance and obesity. Clin Chim Acta 2020; 502: 214–221.
[3] Cunha A, Póvoa AM. Infertility management in women with polycystic ovary syndrome: A review. Porto Biomed J 2021; 6: e116.
[4] Fatemi H, Bilger W, Denis D, Griesinger G, La Marca A, Longobardi S, et al. Dose adjustment of folliclestimulating hormone (FSH) during ovarian stimulation as part of medically-assisted reproduction in clinical studies: A systematic review covering 10 years (2007–2017). J Endocrinol Reprod 2021; 19: 68.
[5] Pedroso DC, Santana VP, Donaires FS, Picinato MC, Giorgenon RC, Santana BA, et al. Telomere length and telomerase activity in immature oocytes and cumulus cells of women with polycystic ovary syndrome. Reprod Sci 2020; 27: 1293–1303.
[6] de Moraes FP, Missio D, Lazzari J, Rovani MT, Ferreira R, Gonçalves PBD, et al. Local regulation of antral follicle development and ovulation in monovulatory species. Anim Reprod 2023; 19: e20220099.
[7] Coxir SA, Costa GMJ, Santos CFd, Alvarenga RdLLS, Lacerda SMdSN. From in vivo to in vitro: Exploring the key molecular and cellular aspects of human female gametogenesis. Hum Cell 2023; 36: 1283–1311.
[8] Kumariya S, Ubba V, Jha RK, Gayen JR. Autophagy in ovary and polycystic ovary syndrome: Role, dispute and future perspective. Autophagy 2021; 17: 2706–2733.
[9] Wang Y, Teng X, Liu J. Research progress on the effect of traditional Chinese medicine on signal pathway related to premature ovarian insufficiency. Evid Based Complement Alternat Med 2022; 2022: 7012978.
[10] Wang Z, Wei H, Wu Z, Zhang X, Sun Y, Gao L, et al. The oocyte cumulus complex regulates mouse sperm migration in the oviduct. Commun Biol 2022; 5: 1327.
[11] Rozés-Salvador V, Siri SO, Musri MM, Conde C. New player in endosomal trafficking: Differential roles of Smad anchor for receptor activation (SARA) protein. Mol Cell Biol 2018; 38: e00446.
[12] Tan Y, Chen Y, Du M, Peng Z, Xie P. USF2 inhibits the transcriptional activity of Smurf1 and SMURF2 to promote breast cancer tumorigenesis. Cell Signal 2019; 53: 49–58.
[13] Rotterdam ESHRE/ASRM-Sponsored PCOS consensus workshop group. Revised 2003 consensus on diagnostic criteria and long-term health risks related to polycystic ovary syndrome (PCOS). Hum Reprod 2004; 19: 41–47.
[14] Gilchrist RB, Smitz J. Oocyte in vitro maturation: Physiological basis and application to clinical practice. Fertil Steril 2023; 119: 524–539.
[15] Li Y, Xiong G, Tan J, Wang S, Wu Q, Wan L, et al. Aberrant activation of the Hedgehog signaling pathway in granulosa cells from patients with polycystic ovary syndrome. Bioengineered 2021; 12: 12123–12134.
[16] Huang R-H, Zhou W-H. Granulosa cell biomarkers to predict oocyte and embryo quality in assisted reproductive technology. Reprod Dev Med 2021; 5: 30–37.
[17] Qin N, Fan X-C, Xu X-X, Tyasi TL, Li S-J, Zhang Y-Y, et al. Cooperative effects of FOXL2 with the members of TGF-?? superfamily on FSH receptor mRNA expression and granulosa cell proliferation from hen prehierarchical follicles. PLoS One 2015; 10: e0141062.
[18] Bai L, Chang HM, Zhang L, Zhu YM, Leung PC. BMP2 increases the production of BDNF through the upregulation of proBDNF and furin expression in human granulosa-lutein cells. FASEB J 2020; 34: 16129–16143.
[19] Tzavlaki K, Moustakas A. TGF-?? Signaling. Biomolecules 2020; 10: 487.
[20] Clark KL, George JW, Przygrodzka E, Plewes MR, Hua G, Wang C, et al. Hippo signaling in the ovary: Emerging roles in development, fertility, and disease. Endocr Rev 2022; 43: 1074–1096.
[21] Gong Z, Yang J, Bai S, Wei S. MicroRNAs regulate granulosa cells apoptosis and follicular development- A review. Asian-Australas J Anim Sci 2020; 33: 1714–1724.
[22] Shi XY, Guan ZQ, Yu JN, Liu HL. Follicle stimulating hormone inhibits the expression of p53 up-regulated modulator of apoptosis induced by reactive oxygen species through PI3K/AKT in mouse granulosa cells. Physiol Res 2020; 69: 687–694.
[23] Shen M, Liu Z, Li B, Teng Y, Zhang J, Tang Y, et al. Involvement of FoxO1 in the effects of follicle-stimulating hormone on inhibition of apoptosis in mouse granulosa cells. Cell Death Dis 2014; 5: e1475.
[24] Bhardwaj JK, Paliwal A, Saraf P, Sachdeva SN. Role of autophagy in follicular development and maintenance of primordial follicular pool in the ovary. J Cell Physiol 2022; 237: 1157–1170.
[25] de Ceuninck van Capelle C, Spit M, Ten Dijke P. Current perspectives on inhibitory Smad7 in health and disease. Crit Rev Biochem Mol 2020; 55: 691–715.
[26] Baddela VS, Michaelis M, Sharma A, Plinski C, Viergutz T, Vanselow J. Estradiol production of granulosa cells is unaffected by the physiological mix of nonesterified fatty acids in follicular fluid. Biol Chem 2022; 298: 691–715.
[27] Du X, Li Q, Yang L, Liu L, Cao Q, Li Q. Smad4 activates Wnt signaling pathway to inhibit granulosa cell apoptosis. Cell Death Dis 2020; 11: 373.
[28] Liu L, Li Q, Yang L, Li Q, Du X. Smad4 feedback activates the canonical TGF-?? family signaling pathways. Int J Mol Sci 2021; 22: 10024.
[29] Piotrowska H, Kempisty B, Sosinska P, Ciesiolka S, Bukowska D, Antosik P, et al. The role of TGF superfamily gene expression in the regulation of folliculogenesis and oogenesis in mammals: a review. Vet Med 2013; 58: 505– 515.
[30] Chandhoke AS, Karve K, Dadakhujaev S, Netherton S, Deng L, Bonni S. The ubiquitin ligase SMURF2 suppresses TGF??-induced epithelial- mesenchymal transition in a sumoylation-regulated manner. Cell Death Differ 2016; 23: 876–888.