Abstract
Gestational diabetes mellitus (GDM) leads to poor pregnancy outcomes. The methods for GDM early diagnosis and treatment are still unknown. This study aimed to investigate the expression and diagnostic potential of miR-34b-3p in GDM patients and further analyzed the effects of miR-34b-3p on HUVECs viability and migration. The expression of miR-34b-3p was detected in HUVECs of GDM and normal pregnant women by qRT-PCR. Then the HUVECs were isolated from normal pregnant women. High glucose (HG) was used to treat the HUVECs to mimic the GDM in vitro. The cell viability and migration were determined by MTT, wound healing assay, and transwell assay. The interaction between miR-34b-3p and PDK1 was evaluated by luciferase activity assay. Our results showed that miR-34b-3p was up-regulated in HUVECs of GDM patients. Then the HUVECs were isolated from normal pregnant women and they were treated with HG to mimic the GDM in vitro. Interestingly, knockdown of miR-34b-3p restored the impairment of HG treatment-induced effects in HUVECs. More importantly, PDK1 was proved to be a potential target of miR-34b-3p. Finally, the rescue experiments confirmed that miR-34b-3p impaired cell viability and migration ability in HUVECs by targeting PDK1. These findings concluded that miR-34b-3p impaired HUVECs viability and migration in GDM by targeting PDK1, which might provide a novel perspective for the pathogenesis and underlying therapeutic target for GDM.
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References
Adameova AD, Bhullar SK, Elimban V, Dhalla NS (2018) Activation of β 1-adrenoceptors may not be involved in arrhythmogenesis in ischemic heart disease. Rev Cardiovasc Med 19:97
Alessi DR, James SR, Downes CP, Holmes AB, Gaffney PR, Reese CB, Cohen P (1997) Characterization of a 3-phosphoinositide-dependent protein kinase which phosphorylates and activates protein kinase Balpha. Curr Biol 7:261–269. https://doi.org/10.1016/s0960-9822(06)00122-9
Bagci et al (2018) Associations between serum levels of adiponectin and resistin and metabolic parameters in pregnant women with gestational diabetes mellitus. Clin Exp Obstet Gynecol 45:539
Ben-Haroush A, Yogev Y, Hod M (2004) Epidemiology of gestational diabetes mellitus and its association with Type 2 diabetes. Diabet Med 21:103–113. https://doi.org/10.1046/j.1464-5491.2003.00985.x
Caporali A et al (2011) Deregulation of microRNA-503 contributes to diabetes mellitus-induced impairment of endothelial function and reparative angiogenesis after limb ischemia. Circulation 123:282–291. https://doi.org/10.1161/CIRCULATIONAHA.110.952325
Cordova-Rivas S et al (2019) 5p and 3p strands of miR-34 family members have differential effects in cell proliferation, migration, and invasion in cervical cancer cells. Int J Mol Sci. https://doi.org/10.3390/ijms20030545
Edu A et al (2016) Placenta changes in pregnancy with gestational diabetes. Rom J Morphol Embryol 57:507–512
Fang LL et al (2017) Potent inhibition of miR-34b on migration and invasion in metastatic prostate cancer cells by regulating the TGF-beta pathway. Int J Mol Sci. https://doi.org/10.3390/ijms18122762
Feng H, Ge F, Du L, Zhang Z, Liu D (2019) MiR-34b-3p represses cell proliferation, cell cycle progression and cell apoptosis in non-small-cell lung cancer (NSCLC) by targeting CDK4. J Cell Mol Med 23:5282–5291. https://doi.org/10.1111/jcmm.14404
Harfouche R, Gratton JP, Yancopoulos GD, Noseda M, Karsan A, Hussain SN (2003) Angiopoietin-1 activates both anti- and proapoptotic mitogen-activated protein kinases. FASEB J 17:1523–1525. https://doi.org/10.1096/fj.02-0698fje
Hashimoto N et al (2006) Ablation of PDK1 in pancreatic beta cells induces diabetes as a result of loss of beta cell mass. Nat Genet 38:589–593. https://doi.org/10.1038/ng1774
Kalimutho M et al (2011) Epigenetically silenced miR-34b/c as a novel faecal-based screening marker for colorectal cancer. Br J Cancer 104:1770–1778. https://doi.org/10.1038/bjc.2011.82
Kim I et al (2000) Angiopoietin-1 induces endothelial cell sprouting through the activation of focal adhesion kinase and plasmin secretion. Circ Res 86:952–959. https://doi.org/10.1161/01.res.86.9.952
Law KP, Zhang H (2017) The pathogenesis and pathophysiology of gestational diabetes mellitus: deductions from a three-part longitudinal metabolomics study in China. Clin Chim Acta 468:60–70. https://doi.org/10.1016/j.cca.2017.02.008
Lawlor MA et al (2002) Essential role of PDK1 in regulating cell size and development in mice. EMBO J 21:3728–3738. https://doi.org/10.1093/emboj/cdf387
Li J et al (2017) miR-26a and miR-26b inhibit esophageal squamous cancer cell proliferation through suppression of c-MYC pathway. Gene 625:1–9. https://doi.org/10.1016/j.gene.2017.05.001
Liu WW, Wang H, Chen XH, Fu SW, Liu ML (2019) miR-34b-3p may promote antiplatelet efficiency of aspirin by inhibiting thromboxane synthase expression. Thromb Haemost 119:1451–1460. https://doi.org/10.1055/s-0039-1692681
Lu TX, Rothenberg ME (2018) MicroRNA. J Allergy Clin Immunol 141:1202–1207. https://doi.org/10.1016/j.jaci.2017.08.034
Mestdagh P, Van Vlierberghe P, De Weer A, Muth D, Westermann F, Speleman F, Vandesompele J (2009) A novel and universal method for microRNA RT-qPCR data normalization. Genome Biol 10:R64. https://doi.org/10.1186/gb-2009-10-6-r64
Miao M, Dai M, Zhang Y, Sun F, Guo X, Sun G (2017) Influence of maternal overweight, obesity and gestational weight gain on the perinatal outcomes in women with gestational diabetes mellitus. Sci Rep 7:305. https://doi.org/10.1038/s41598-017-00441-z
Mizuno K et al (2017) The microRNA expression signature of small cell lung cancer: tumor suppressors of miR-27a-5p and miR-34b-3p and their targeted oncogenes. J Human Genet 62:671–678. https://doi.org/10.1038/jhg.2017.27
Mora A, Komander D, van Aalten DM, Alessi DR (2004) PDK1, the master regulator of AGC kinase signal transduction. Sem Cell Dev Biol 15:161–170. https://doi.org/10.1016/j.semcdb.2003.12.022
Muralimanoharan S, Maloyan A, Myatt L (2016) Mitochondrial function and glucose metabolism in the placenta with gestational diabetes mellitus: role of miR-143. Clin Sci 130:931–941. https://doi.org/10.1042/CS20160076
Papapetropoulos A, Garcia-Cardena G, Dengler TJ, Maisonpierre PC, Yancopoulos GD, Sessa WC (1999) Direct actions of angiopoietin-1 on human endothelium: evidence for network stabilization, cell survival, and interaction with other angiogenic growth factors. Lab Investig 79:213–223
Papapetropoulos A et al (2000) Angiopoietin-1 inhibits endothelial cell apoptosis via the Akt/survivin pathway. J Biol Chem 275:9102–9105. https://doi.org/10.1074/jbc.275.13.9102
Peng HY, Li HP, Li MQ (2018) High glucose induces dysfunction of human umbilical vein endothelial cells by upregulating miR-137 in gestational diabetes mellitus. Microvasc Res 118:90–100. https://doi.org/10.1016/j.mvr.2018.03.002
Peng HY, Li MQ, Li HP (2019) MiR-137 restricts the viability and migration of HTR-8/SVneo cells by downregulating FNDC5 in gestational diabetes mellitus. Curr Mol Med 19:494–505. https://doi.org/10.2174/1566524019666190520100422
Primo L, di Blasio L, Roca C, Droetto S, Piva R, Schaffhausen B, Bussolino F (2007) Essential role of PDK1 in regulating endothelial cell migration. J Cell Biol 176:1035–1047. https://doi.org/10.1083/jcb.200607053
Shi Z, Zhao C, Guo X, Ding H, Cui Y, Shen R, Liu J (2014) Differential expression of microRNAs in omental adipose tissue from gestational diabetes mellitus subjects reveals miR-222 as a regulator of ERalpha expression in estrogen-induced insulin resistance. Endocrinology 155:1982–1990. https://doi.org/10.1210/en.2013-2046
Spaight C, Gross J, Horsch A, Puder JJ (2016) Gestational Diabetes Mellitus Endocrine development 31:163–178. https://doi.org/10.1159/000439413
Sultan SA, Liu W, Peng Y, Roberts W, Whitelaw D, Graham AM (2015) The role of maternal gestational diabetes in inducing fetal endothelial dysfunction. J Cell Physiol 230:2695–2705. https://doi.org/10.1002/jcp.24993
Tan Y, Zhang T, Zhou L, Liu S, Liang C (2019) MiR-34b-3p represses the multidrug-chemoresistance of bladder cancer cells by regulating the CCND2 and P2RY1 genes. Med Sci Monit 25:1323–1335. https://doi.org/10.12659/MSM.913746
Tanase-Nakao K, Arata N, Kawasaki M, Yasuhi I, Sone H, Mori R, Ota E (2017) Potential protective effect of lactation against incidence of type 2 diabetes mellitus in women with previous gestational diabetes mellitus: a systematic review and meta-analysis. Diabetes. https://doi.org/10.1002/dmrr.2875
Trout KK, Averbuch T, Barowski M (2011) Promoting breastfeeding among obese women and women with gestational diabetes mellitus. Curr Diabet Rep 11:7–12. https://doi.org/10.1007/s11892-010-0159-6
Tryggestad JB et al (2016) Influence of gestational diabetes mellitus on human umbilical vein endothelial cell miRNA. Clin Sci 130:1955–1967. https://doi.org/10.1042/CS20160305
Wang P, Wang H, Li C, Zhang X, Xiu X, Teng P, Wang Z (2019) Dysregulation of microRNA-657 influences inflammatory response via targeting interleukin-37 in gestational diabetes mellitus. J Cell Physiol 234:7141–7148. https://doi.org/10.1002/jcp.27468
Westgate JA et al (2006) Hyperinsulinemia in cord blood in mothers with type 2 diabetes and gestational diabetes mellitus in New Zealand. Diabet Care 29:1345–1350. https://doi.org/10.2337/dc05-1677
Yancopoulos GD, Davis S, Gale NW, Rudge JS, Wiegand SJ, Holash J (2000) Vascular-specific growth factors and blood vessel formation. Nature 407:242–248. https://doi.org/10.1038/35025215
Zheng S et al (2015) Ang-(1–7) promotes the migration and invasion of human renal cell carcinoma cells via Mas-mediated AKT signaling pathway. Biochem Biophys Res Commun 460:333–340. https://doi.org/10.1016/j.bbrc.2015.03.035
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FS and AC designed the study, supervised the data collection, analyzed the data; QY and XC interpreted the data and prepared the manuscript for publication; LL and XH supervised the data collection, analyzed the data, and reviewed the draft of the manuscript. All the authors have read and approved the manuscript.
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Song, F., Cai, A., Ye, Q. et al. MiR-34b-3p Impaired HUVECs Viability and Migration via Targeting PDK1 in an In Vitro Model of Gestational Diabetes Mellitus. Biochem Genet 59, 1381–1395 (2021). https://doi.org/10.1007/s10528-021-10064-9
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DOI: https://doi.org/10.1007/s10528-021-10064-9