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Testosterone promotes human foreskin fibroblast growth through miR-143-3p targeting IGFBP-3

  • Qian-Long Peng1
  • Yao-Wang Zhao1
  • Wen Tian1,*,

1Department of Urology, Hunan Children’s Hospital, 410007 Changsha, Hunan, China

DOI: 10.22514/jomh.2023.082 Vol.19,Issue 9,September 2023 pp.15-25

Submitted: 18 November 2022 Accepted: 25 May 2023

Published: 30 September 2023

*Corresponding Author(s): Wen Tian E-mail:


Testosterone is an important male hormone, which could improve the maintenance and recovery of gonadal function in males as well as the repair of human hypospadias and cell fibrosis. Our study focused on investigating the regulatory effect of testosterone in human foreskin fibroblasts (HFF-1) and regulatory mechanisms involved. In this study, HFF-1 cells were treated with testosterone, and cell viability and migration were assessed by cell counting kit-8 (CCK8) and Transwell assays. The expression levels of androgen receptor (AR), miR-143-3p and insulin-like growth factor binding protein-3 (IGFBP-3) were measured by quantitative real-time PCR (qRT-PCR), Western blotting, and immunofluorescence. In addition, a potential binding site for miR-143-3p on IGFBP-3 was predicted and its direct binding was further confirmed by a dual luciferase reporter assay. These results showed that testosterone increased the viability and migration of HFF-1 cells. Testosterone could down-regulate miR-143-3p and up-regulate IGFBP-3 and AR. Overexpression of miR-143-3p hindered HFF-1 cell viability and negatively regulated IGFBP-3, whereas inhibition of IGFBP-3 impeded cell viability and migration. Furthermore, miR-143-3p was found to directly bind to IGFBP-3. Overexpression of IGFBP-3 countered the regulation of HFF-1 cells by miR-143-3p mimics. In conclusion, this study showed that testosterone promoted the proliferation and migration of HFF-1 cells and AR signaling, at least via the miR-143-3p/IGFBP-3 axis. This discovery presents a novel insight for testosterone application in male disorders like hypospadias.


Testosterone; miR-143-3p; IGFBP-3; HFF-1 cells

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Qian-Long Peng,Yao-Wang Zhao,Wen Tian. Testosterone promotes human foreskin fibroblast growth through miR-143-3p targeting IGFBP-3. Journal of Men's Health. 2023. 19(9);15-25.


[1] Ceccarelli PL, Lucaccioni L, Poluzzi F, Bianchini A, Biondini D, Iughetti L, et al. Hypospadias: clinical approach, surgical technique and long-term outcome. BMC Pediatrics. 2021; 21: 523.

[2] Gul M, Hildorf S, Silay MS. Sexual functions and fertility outcomes after hypospadias repair. International Journal of Impotence Research. 2021; 33: 149–163.

[3] Kaefer M, Rink R, Misseri R, Winchester P, Proctor C, Ben Maamar M, et al. Role of epigenetics in the etiology of hypospadias through penile foreskin DNA methylation alterations. Scientific Reports. 2023; 13: 555.

[4] Qin XY, Kojima Y, Mizuno K, Ueoka K, Muroya K, Miyado M, et al. Identification of novel low-dose bisphenol a targets in human foreskin fibroblast cells derived from hypospadias patients. PLOS ONE. 2012; 7: e36711.

[5] Wittert G, Grossmann M. Obesity, type 2 diabetes, and testosterone in ageing men. Reviews in Endocrine and Metabolic Disorders. 2022; 23: 1233–1242.

[6] Wang C, Swerdloff RS. Testosterone replacement therapy in hypogonadal men. Endocrinology and Metabolism Clinics of North America. 2022; 51: 77–98.

[7] Auerbach JM, Khera M. Testosterone replacement therapy and cardio-vascular disease. International Journal of Impotence Research. 2022; 34: 685–690.

[8] Jin B, An H. Baicalin alleviates benign prostate hyperplasia through androgen-dependent apoptosis. Aging. 2020; 12: 2142–2155.

[9] Jang YJ, Jung HY, Myeong JY, Song KH, Kwon J, Kim D, et al. Effects of alginate oligosaccharide on testosterone-induced benign prostatic hyperplasia in orchiectomized rats. Nutrients. 2023; 15: 682.

[10] Godlewski KF, Mittal S, Hyacinthe N, Fischer K, Weaver J, Van Batavia J, et al. Does preoperative testosterone administration decrease complications in distal hypospadias repair with urethroplasty? The Journal of Urology. 2023. [Preprint].

[11] Stern JM, Chen J, Peters SB, Stahl PJ, El-Chaar M, Felsen D, et al. Testosterone treatment of human foreskin in a novel transplant model. Urology. 2004; 63: 999–1003.

[12] Hussen BM, Hidayat HJ, Salihi A, Sabir DK, Taheri M, Ghafouri-Fard S. MicroRNA: a signature for cancer progression. Biomedicine & Pharmacotherapy. 2021; 138: 111528.

[13] Michlewski G, Cáceres JF. Post-transcriptional control of miRNA biogenesis. RNA. 2019; 25: 1–16.

[14] Xu X, Guan R, Gong K, Xie H, Shi L. Circ_FURIN knockdown assuages testosterone-induced human ovarian granulosa-like tumor cell disorders by sponging miR-423-5p to reduce MTM1 expression in polycystic ovary syndrome. Reproductive Biology and Endocrinology. 2022; 20: 32.

[15] Kalinina T, Kononchuk V, Alekseenok E, Abdullin G, Sidorov S, Ovchinnikov V, et al. Associations between the levels of estradiol-, progesterone-, and testosterone-sensitive MiRNAs and main clinico-pathologic features of breast cancer. Journal of Personalized Medicine. 2021; 12: 4.

[16] Wang Y, Li H, Shi Y, Wang S, Xu Y, Li H, et al. MiR-143-3p impacts on pulmonary inflammatory factors and cell apoptosis in mice with mycoplasmal pneumonia by regulating TLR4/MyD88/NF-κB pathway. Bioscience Reports. 2020; 40: BSR20193419.

[17] Jiang B, Yuan C, Han J, Shen M, Zhou X, Zhou L. MiR-143-3p inhibits the differentiation of osteoclast induced by synovial fibroblast and monocyte coculture in adjuvant-induced arthritic rats. BioMed Research International. 2021; 2021: 1–10.

[18] Tang J, Pan H, Wang W, Qi C, Gu C, Shang A, et al. MiR-495-3p and miR-143-3p co-target CDK1 to inhibit the development of cervical cancer. Clinical and Translational Oncology. 2021; 23: 2323–2334.

[19] Zhang G, Liu Z, Zhong J, Lin L. Circ‐ACAP2 facilitates the progression of colorectal cancer through mediating miR‐143‐3p/FZD4 axis. European Journal of Clinical Investigation. 2021; 51: e13607.

[20] Zhang L, Jiang H, Zhang Y, Wang C, Xia X, Sun Y. GR silencing impedes the progression of castration-resistant prostate cancer through the JAG1/NOTCH2 pathway via up-regulation of microRNA-143-3p. Cancer Biomarkers. 2020; 28: 483–497.

[21] Bonifacio LN, Jarstfer MB. MiRNA profile associated with replicative senescence, extended cell culture, and ectopic telomerase expression in human foreskin fibroblasts. PLOS ONE. 2010; 5: e12519.

[22] Song F, Zhou X, Hu Y, Li G, Wang Y. The roles of insulin-like growth factor binding protein family in development and diseases. Advances in Therapy. 2021; 38: 885–903.

[23] Song B, Xu J, Zhong P, Fang L. MiR-125a-5p silencing inhibits cerebral ischemia-induced injury through targeting IGFBP3. Folia Neuropathologica. 2021; 59: 121–130.

[24] Feng N, Wang Z, Wu Y, Zheng H, Jiang X, Wang Z, et al. ADAMTS9-as2 promotes angiogenesis of brain microvascular endothelial cells through regulating miR-185-5p/IGFBP-2 axis in ischemic stroke. Molecular Neurobiology. 2022; 59: 2593–2604.

[25] Kerr A, Baxter RC. Noncoding RNA actions through IGFs and IGF binding proteins in cancer. Oncogene. 2022; 41: 3385–3393.

[26] Jogie-Brahim S, Feldman D, Oh Y. Unraveling insulin-like growth factor binding protein-3 actions in human disease. Endocrine Reviews. 2009; 30: 417–437.

[27] Shih H, Chen C, Torng P. IGFBP3 inhibits angiogenesis through intracellular regulation of THBS1 expression. American Journal of Cancer Research. 2020; 10: 1728–1744.

[28] Hu D, Ge Y, Cui Y, Li K, Chen J, Zhang C, et al. Upregulated IGFBP3 with aging is involved in modulating apoptosis, oxidative stress, and fibrosis: a target of age-related erectile dysfunction. Oxidative Medicine and Cellular Longevity. 2022; 2022: 1–18.

[29] Chen X, Shao Y, Wei W, Shen H, Li Y, Chen Y, et al. Downregulation of LOX promotes castration-resistant prostate cancer progression via IGFBP3. Journal of Cancer. 2021; 12: 7349–7357.

[30] Yoshizawa A. Testosterone and insulin-like growth factor (IGF) I interact in controlling IGF-binding protein production in androgen-responsive foreskin fibroblasts. Journal of Clinical Endocrinology & Metabolism. 2000; 85: 1627–1633.

[31] Ouyang X, Feng L, Liu G, Yao L, Wang Z, Liu S, et al. Androgen receptor (AR) decreases HCC cells migration and invasion via miR-325/ACP5 signaling. Journal of Cancer. 2021; 12: 1915–1925.

[32] Chen Q, Zhang H, Yang Y, Zhang S, Wang J, Zhang D, et al. Metformin attenuates UVA-induced skin photoaging by suppressing mitophagy and the PI3K/AKT/mTOR pathway. International Journal of Molecular Sciences. 2022; 23: 6960.

[33] Liu X, Song YJ, Chen X, Huang MY, Zhao CX, Zhou X, et al. Asiaticoside combined with carbon ion implantation to improve the biocompatibility of silicone rubber and to reduce the risk of capsule contracture. Frontiers in Bioengineering and Biotechnology. 2022; 10: 810244.

[34] Wang BJ, Huang SH, Kao CL, Muller CJF, Wang YP, Chang KH, et al. Aspalathus linearis suppresses cell survival and proliferation of enzalutamide-resistant prostate cancer cells via inhibition of c-Myc and stability of androgen receptor. PLOS ONE. 2022; 17: e0270803.

[35] Ding M, Jiang C, Zhang Y, Zhao J, Han B, Xia S. SIRT7 depletion inhibits cell proliferation and androgen-induced autophagy by suppressing the AR signaling in prostate cancer. Journal of Experimental & Clinical Cancer Research. 2020; 39: 28.

[36] Ma L, Dong L, Zhu J, Yu J, Deng Q. Exploration of potential therapeutic targets for stroke based on the GEO database. Annals of Translational Medicine. 2021; 9: 1759–1759.

[37] Liang D, Tian C, Zhang X. lncRNA MNX1‑AS1 promotes prostate cancer progression through regulating miR-2113/MDM2 axis. Molecular Medicine Reports. 2022; 26: 231.

[38] Lucas-Herald AK, Montezano AC, Alves-Lopes R, Haddow L, Al-imussina M, O’Toole S, et al. Vascular dysfunction and increased cardiovascular risk in hypospadias. European Heart Journal. 2022; 43: 1832–1845.

[39] Jamroze A, Chatta G, Tang DG. Androgen receptor (AR) heterogeneity in prostate cancer and therapy resistance. Cancer Letters. 2021; 518: 1–9.

[40] Chao chen, Jian Y, Zhao X, Liu Y, Xie Q. The involvement of hsa_circ_0000417 in the development of hypospadias by regulating AR. Differentiation. 2020; 116: 9–15.

[41] Vottero A, Minari R, Viani I, Tassi F, Bonatti F, Neri TM, et al. Evidence for epigenetic abnormalities of the androgen receptor gene in foreskin from children with hypospadias. The Journal of Clinical Endocrinology & Metabolism. 2011; 96: E1953–E1962.

[42] Yang C, Xu X, Lin P, Luo B, Luo S, Huang H, et al. Overexpression of long noncoding RNA MCM3AP-as1 promotes osteogenic differentiation of dental pulp stem cells via miR-143-3p/IGFBP5 axis. Human Cell. 2022; 35: 150–162.

[43] An N, Peng J, He G, Fan X, Li F, Chen H. Involvement of activation of mitogen-activated protein kinase (MAPK)/Extracellular signal-regulated kinase (ERK) signaling pathway in proliferation of urethral plate fibroblasts in finasteride-induced rat hypospadias. Medical Science Monitor. 2018; 24: 8984–8992.

[44] Di Lodovico E, Facondo P, Delbarba A, Pezzaioli LC, Maffezzoni F, Cappelli C, et al. Testosterone, hypogonadism, and heart failure. Circulation: Heart Failure. 2022; 15: e008755.

[45] Jayasena CN, Anderson RA, Llahana S, Barth JH, MacKenzie F, Wilkes S, et al. Society for endocrinology guidelines for testosterone replacement therapy in male hypogonadism. Clinical Endocrinology. 2022; 96: 200–219.

[46] Garibotto G, Esposito P, Picciotto D, Verzola D. Testosterone disorders and male hypogonadism in kidney disease. Seminars in Nephrology. 2021; 41: 114–125.

[47] Corona G, Maggi M. The role of testosterone in male sexual function. Reviews in Endocrine and Metabolic Disorders. 2022; 23: 1159–1172.

[48] Anawalt BD, Matsumoto AM. Aging and androgens: physiology and clinical implications. Reviews in Endocrine and Metabolic Disorders. 2022; 23: 1123–1137.

[49] Barbonetti A, D’Andrea S, Francavilla S. Testosterone replacement therapy. Andrology. 2020; 8: 1551–1566.

[50] Wong NC, Braga LH. The influence of pre-operative hormonal stimula-tion on hypospadias repair. Frontiers in Pediatrics. 2015; 3: 31.

[51] Mittal S, Eftekharzadeh S, Christianson SS, Hyacinthe N, Tan C, Weiss DA, et al. Quantifying glans width changes in response to preoperative androgen stimulation in patients undergoing hypospadias repair. The Journal of Urology. 2022; 207: 1314–1321.

[52] Chen Z, Lin X, Wang Y, Xie H, Chen F. Dysregulated expression of androgen metabolism genes and genetic analysis in hypospadias. Molecular Genetics & Genomic Medicine. 2020; 8: e1346.

[53] Richard MA, Sok P, Canon S, Nembhard WN, Brown AL, Peckham-Gregory EC, et al. Altered mechanisms of genital development identified through integration of DNA methylation and genomic measures in hypospadias. Scientific Reports. 2020; 10: 12715.

[54] Taghavi K, O’Hagan LA, Hewitt JK, Mouriquand PD. Defining the role of pre-operative hormonal therapy in hypospadias. Journal of Paediatrics and Child Health. 2022; 58: 1508–1519.

[55] Huang Y, Yan H, Yang Y, Zhou J, Xu Q, Meng H. Downregulated miR-181a alleviates H2O2-induced oxidative stress and cellular senescence by targeting PDIA6 in human foreskin fibroblasts. Anais Brasileiros de Dermatologia. 2023; 98: 17–25.

[56] Mudali S. Effects of testosterone on body composition of the aging male. Mechanisms of Ageing and Development. 2004; 125: 297–304.

[57] Wang R, Zhang H, Ding W, Fan Z, Ji B, Ding C, et al. miR-143 promotes angiogenesis and osteoblast differentiation by targeting HDAC7. Cell Death & Disease. 2020; 11: 179.

[58] Chodari L. The effect of testosterone and voluntary exercise, alone or together, on miRNA-126 expression changes in heart of diabetic rats. Acta Endocrinologica. 2017; 13: 266–271.

[59] Rooman RPA, De Beeck LO, Martin M, van Doorn J, Mohan S, Du Caju MVL. Ethinylestradiol and testosterone have divergent effects on circulating IGF system components in adolescents with constitutional tall stature. European Journal of Endocrinology. 2005; 152: 597–604.

[60] Long Z, Gong F, Li Y, Fan Z, Li J. Circ_0000285 regulates prolifer-ation, migration, invasion and apoptosis of osteosarcoma by miR-409-3p/IGFBP3 axis. Cancer Cell International. 2020; 20: 481.

[61] Zielinska HA, Daly CS, Alghamdi A, Bahl A, Sohail M, White P, et al. Interaction between GRP78 and IGFBP-3 affects tumourigenesis and prognosis in breast cancer patients. Cancers. 2020; 12: 3821.

[62] Cai Q, Dozmorov M, Oh Y. IGFBP-3/IGFBP-3 receptor system as an anti-tumor and anti-metastatic signaling in cancer. Cells. 2020; 9: 1261.

[63] Li C, Liu B, Wang Z, Xie F, Qiao W, Cheng J, et al. Salvianolic acid B improves myocardial function in diabetic cardiomyopathy by suppressing IGFBP3. Journal of Molecular and Cellular Cardiology. 2020; 139: 98–112.

[64] Tao A, Wang X, Li C. Effect of lycopene on oral squamous cell carcinoma cell growth by inhibiting IGF1 pathway. Cancer Management and Research. 2021; 13: 723–732.

[65] Chen S, Tang Y, Liu Y, Zhang P, Lv L, Zhang X, et al. Exosomes derived from miR-375-overexpressing human adipose mesenchymal stem cells promote bone regeneration. Cell Proliferation. 2019; 52: e12669.

[66] Lv M, Zhou L, Ge P, Li Y, Zhang J, Zhou D. Over‐expression of hsa_circ_0000116 in patients with non‐obstructive azoospermia and its predictive value in testicular sperm retrieval. Andrology. 2020; 8: 1834–1843.

[67] Liu M, Liu Y, Pei L, Zhang Q, Xiao H, Chen Y, et al. Prenatal dexamethasone exposure programs the decreased testosterone synthesis in offspring rats by low level of endogenous glucocorticoids. Acta Pharmacologica Sinica. 2022; 43: 1461–1472.

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