High glucose suppresses the viability and proliferation of HTR‑8/SVneo cells through regulation of the miR‑137/PRKAA1/IL‑6 axis

Peng,Hai‑Yan; Li,Ming‑Qing; Li,Hua‑Ping

doi:10.3892/ijmm.2018.3686

High glucose suppresses the viability and proliferation of HTR‑8/SVneo cells through regulation of the miR‑137/PRKAA1/IL‑6 axis

Authors:
- Hai‑Yan Peng
- Ming‑Qing Li
- Hua‑Ping Li
View Affiliations

Affiliations: Department of Gynecology and Obstetrics, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai 200233, P.R. China, Laboratory for Reproductive Immunology, Shanghai Key Laboratory of Female Reproductive Endocrine Related Diseases, Obstetrics and Gynecology Hospital of Fudan University, Shanghai 200011, P.R. China
Published online on: May 17, 2018 https://doi.org/10.3892/ijmm.2018.3686
Pages: 799-810
Copyright: © Peng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The aim of the present study was to investigate the mechanism underlying the high glucose (HG)‑associated regulation of HTR‑8/SVneo cell viability and proliferation during gestational diabetes mellitus (GDM), and to verify the association of microRNA (miR)‑137, protein kinase AMP‑activated catalytic subunit α1 (PRKAA1) and interlukin‑6 (IL‑6). miR‑137‑overexpressing and negative control HTR‑8/SVneo cells were established by lentiviral vector infection. Cell Counting Kit‑8 and colony formation assays were used to analyze the viability and proliferation of HTR‑8/SVneo cells. Reverse transcription‑quantitative polymerase chain reaction analysis was used to determine the transcriptional activity of miR‑137, PRKAA1 and Il‑6, and ELISA and western blot analysis were used to measure the protein levels of IL‑6 and PRKAA1, respectively. It was demonstrated that PRKAA1 was decreased in the placental tissues of women with GDM and HG‑treated HTR‑8/SVneo cells, and that HG upregulated miR‑137 and IL‑6 in trophoblasts. The overexpression of miR‑137 decreased levels of PRKAA1 and increased levels of IL‑6 in the HTR‑8/SVneo cells. An inhibitor of PRKAA1 promoted the secretion of IL‑6, whereas an agonist of PRKAA1 suppressed the production of IL‑6. HG treatment and the overexpression of miR‑137 reduced the viability and proliferation of HTR‑8/SVneo cells in vitro, whereas the activation of PRKAA1 or incubation with IL‑6 antibody reversed these effects. Overall, it was concluded that HG suppressed the viability and proliferation of trophoblast cells through the miR‑137/PRKAA1/IL‑6 axis, which may contribute to pathological changes of placental tissues in GDM.

View Figures

View References

1	Kleinwechter H and Demandt N: Diabetes in pregnancy-type 1/type 2 diabetes mellitus and gestational diabetes mellitus. Dtsch Med Wochenschr. 141:1296–1303. 2016.In German. PubMed/NCBI
2	Bánhidy F, Acs N, Puhó EH and Czeizel AE: Congenital abnormalities in the offspring of pregnant women with type 1, type 2 and gestational diabetes mellitus: A population-based case-control study. Congenit Anom. 50:115–121. 2010. View Article : Google Scholar
3	Callec R, Perdriolle-Galet E, Sery GA and Morel O: Type 2 diabetes in pregnancy: Rates of fetal malformations and level of preconception care. J Obstet Gynaecol. 34:648–649. 2014. View Article : Google Scholar : PubMed/NCBI
4	Kaiser J: Gearing up for a closer look at the human placenta. Science. 344:10732014. View Article : Google Scholar
5	Zong S, Li C, Luo C, Zhao X, Liu C, Wang K, Jia W, Bai M, Yin M, Bao S, et al: Dysregulated expression of IDO may cause unexplained recurrent spontaneous abortion through suppression of trophoblast cell proliferation and migration. Sci Rep. 27:199162016. View Article : Google Scholar
6	Tian FJ, Qin CM, Li XC, Wu F, Liu XR, Xu WM and Lin Y: Decreased stathmin-1 expression inhibits trophoblast proliferation and invasion and is associated with recurrent miscarriage. Am J Pathol. 185:2709–2721. 2015. View Article : Google Scholar : PubMed/NCBI
7	Liong S and Lappas M: Activation of AMPK improves inflammation and insulin resistance in adipose tissue and skeletal muscle from pregnant women. J Physiol Biochem. 71:703–717. 2015. View Article : Google Scholar : PubMed/NCBI
8	Li J and Li J, Wei T and Li J: Down-regulation of MicroRNA-137 improves high glucose induced oxidative stress injury in human umbilical vein endothelial cells by up-regulation of AMPKα1. Cell Physiol Biochem. 39:847–859. 2016. View Article : Google Scholar
9	Yao G, Zhang Y, Wang D, Yang R, Sang H, Han L, Zhu Y, Lu Y, Tan Y and Shang Z: GDM-induced macrosomia is reversed by Cav-1 via AMPK-mediated fatty acid transport and GLUT1-mediated glucose transport in placenta. PLoS One. 12:e01704902017. View Article : Google Scholar : PubMed/NCBI
10	Chen H, Lan HY, Roukos DH and Cho WC: Application of microRNAs in diabetes mellitus. J Endocrinol. 222:R1–R10. 2014. View Article : Google Scholar : PubMed/NCBI
11	Haertle L, El Hajj N, Dittrich M, Müller T, Nanda I, Lehnen H and Haaf T: Epigenetic signatures of gestational diabetes mellitus on cord blood methylation. Clin Epigenetics. 9:282017. View Article : Google Scholar : PubMed/NCBI
12	Ornoy A, Reece EA, Pavlinkova G, Kappen C and Miller RK: Effect of maternal diabetes on the embryo, fetus, and children: Congenital anomalies, geneticand epigenetic changes and developmental outcomes. Birth Defects Res C Embryo Today. 105:53–72. 2015. View Article : Google Scholar : PubMed/NCBI
13	Li J, Song L, Zhou L, Wu J, Sheng C, Chen H, Liu Y, Gao S and Huang W: A MicroRNA signature in gestational diabetes mellitus associated with risk of macrosomia. Cell Physiol Biochem. 37:243–252. 2015. View Article : Google Scholar : PubMed/NCBI
14	Lu TM, Lu W and Zhao LJ: MicroRNA-137 affects proliferation and migration of placenta trophoblast cells in preeclampsia by targeting ERRα. Reprod Sci. pii: 1933719116650754. 2016.
15	Lappas M: Double stranded viral RNA induces inflammation and insulin resistance in skeletal muscle from pregnant women in vitro. Metabolism. 64:642–653. 2015. View Article : Google Scholar : PubMed/NCBI
16	Kim SY, Jeong S, Jung E, Baik KH, Chang MH, Kim SA, Shim JH, Chun E and Lee KY: AMP-activated protein kinase-α1 as an activating kinase of TGF-β-activated kinase 1 has a key role in inflammatory signals. Cell Death Dis. 3:e3572012. View Article : Google Scholar
17	Lim R, Barker G and Lappas M: Activation of AMPK in human fetal membranes alleviates infection-induced expression of pro-inflammatory and pro-labour mediators. Placenta. 36:454–462. 2015. View Article : Google Scholar : PubMed/NCBI
18	Huang L, Chiang SH, Hsueh CH, Liang YJ and Chen YJ: Lai LP. Metformin inhibits TNF-alpha-induced Ikappa-B kinase phosphorylation, IkappaB-alpha degradation and IL-6 production in endothelial cells through I3K-dependent AMPK phosphorylation. Int J Cardiol. 134:169–175. 2009. View Article : Google Scholar
19	American Diabetes Association: Diagnosis and classification of diabetes mellitus: Diabetes Care. 35(Suppl 1): S64–S71. 2012. View Article : Google Scholar
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔC T method. Methods. 25:402–408. 2001. View Article : Google Scholar
21	Daniele G, Guardado Mendoza R, Winnier D, Fiorentino TV, Pengou Z, Cornell J, Andreozzi F, Jenkinson C, Cersosimo E, Federici M, et al: The inflammatory status score including IL-6, TNF-α, osteopontin, fractalkine, MCP-1 and adiponectin underlies whole-body insulin resistance and hyperglycemia in type 2 diabetes mellitus. Acta Diabetol. 51:123–131. 2014. View Article : Google Scholar
22	Bigham AW, Julian CG, Wilson MJ, Vargas E, Browne VA, Shriver MD and Moore LG: Maternal RKAA1 and EDNRA genotypes are associated with birth weight, and RKAA1 with uterine artery diameter and metabolic homeostasis at high altitude. Physiol Genomics. 46:687–697. 2014. View Article : Google Scholar : PubMed/NCBI
23	He C, Li H, Viollet B, Zou MH and Xie Z: AMPK suppresses vascular inflammation in vivo by inhibiting signal transducer and activator of transcription-1. Diabetes. 64:4285–4297. 2015. View Article : Google Scholar : PubMed/NCBI
24	Visiedo F, Bugatto F, Sánchez V, Cózar-Castellano I, Bartha JL and Perdomo G: High glucose levels reduce fatty acid oxidation and increase triglyceride accumulation in human placenta. Am J Physiol Endocrinol Metab. 305:E205–E212. 2013. View Article : Google Scholar : PubMed/NCBI
25	Wang LF, Wang HJ, Ao D, Liu Z, Wang Y and Yang HX: Influence of pre-pregnancy obesity on the development of macrosomia and large for gestational age in women with or without gestational diabetes mellitus in Chinese population. J Perinatol. 35:985–990. 2015. View Article : Google Scholar : PubMed/NCBI
26	Jarmuzek P, Wielgos M and Bomba-Opon D: Placental pathologic changes in gestational diabetes mellitus. Neuro Endocrinol Lett. 36:101–105. 2015.PubMed/NCBI
27	Leng J, Li W, Zhang S, Liu H, Wang L, Liu G, Li N, Redman LM, Baccarelli AA, Hou L and Hu G: GDM women's pre-pregnancy overweight/obesity and gestational weight gain on offspring overweight status. PLoS One. 10:e01295362015. View Article : Google Scholar : PubMed/NCBI
28	Sedlic F, Muravyeva MY, Sepac A, Sedlic M, Williams AM, Yang M, Bai X and Bosnjak ZJ: Targeted modification of mitochondrial ROS production converts high glucose-induced cytotoxicity to cytoprotection: Effects on anesthetic preconditioning. J Cell Physiol. 232:216–224. 2017. View Article : Google Scholar
29	Kuricová K, Pácal L, Šoupal J, Prázný M and Kaňková K: Effect of glucose variability on pathways associated with glucotoxicity in diabetes: Evaluation of a novel in vitro experimental approach. Diabetes Res Clin Pract. 114:1–8. 2016. View Article : Google Scholar : PubMed/NCBI
30	Liu J, Chen S, Ren W, Liu J, Yang P, Chen Z, Zhang Q and Yang F: Lipopolysaccharide-induced suppression of periodontal ligament cell proliferation and apoptosis are strengthened under high glucose conditions. Arch Oral Biol. 79:70–76. 2017. View Article : Google Scholar : PubMed/NCBI
31	Abdallah AM, Alves BC, Gehrke FS, Sant' Ana AV, Azzalis LA, Junqueira VB, Adami F, Pereira EC and Fonseca FL: Inability of turbidimetry method in detecting glycated hemoglobin to select diabetes mellitus patients according to their concentrations of blood glucose levels. J Clin Lab Anal. 29:312–316. 2015. View Article : Google Scholar
32	Pedersen ML, Olesen J, Jørgensen ME and Damm P: Gestational diabetes mellitus in Greenland: A national study of prevalence and testing efficacy. Int J Circumpolar Health. 75:321672016. View Article : Google Scholar : PubMed/NCBI
33	Wojtaszewski JF, Birk JB, Frøsig C, Holten M, Pilegaard H and Dela F: 5′AMP activated protein kinase expression in human skeletal muscle: Effects of strength training and type 2 diabetes. J Physiol. 564:563–573. 2005. View Article : Google Scholar : PubMed/NCBI
34	Song YM, Lee YH, Kim JW, Ham DS, Kang ES, Cha BS, Lee HC and Lee BW: Metformin alleviates hepatosteatosis by restoring SIRT1-mediated autophagy induction via an AMP-activated protein kinase-independent pathway. Autophagy. 11:46–59. 2015. View Article : Google Scholar :
35	Waker CA, Albers RE, Pye RL, Doliboa SR, Wyatt CN, Brown TL and Mayes DA: AMPK knockdown in placental labyrinthine progenitor cells results in restriction of critical energy resources and terminal differentiation failure. Stem Cells Dev. 26:808–817. 2017. View Article : Google Scholar : PubMed/NCBI
36	Zhang L, Han YJ, Zhang X, Wang X, Bao B, Qu W and Liu J: Luteolin reduces obesity-associated insulin resistance in mice by activating ampkα1 signaling in adipose tissue macrophages. Diabetologia. 59:2219–2228. 2016. View Article : Google Scholar : PubMed/NCBI
37	Floris I, Descamps B, Vardeu A, Mitić T, Posadino AM, Shantikumar S, Sala-Newby G, Capobianco G, Mangialardi G, Howard L, et al: Gestational diabetes mellitus impairs fetal endothelial cell functions through a mechanism involving microRNA-101 and histone methyltransferase enhancer of zester homolog-2. Arterioscler Thromb Vasc Biol. 35:664–674. 2015. View Article : Google Scholar : PubMed/NCBI
38	Dang J, Bian YQ, Sun JY, Chen F, Dong GY, Liu Q, Wang XW, Kjems J, Gao S and Wang QT: MicroRNA-137 promoter methylation in oral lichen planus and oral squamous cell carcinoma. J Oral Pathol Med. 42:315–321. 2013. View Article : Google Scholar
39	Langevin SM, Stone RA, Bunker CH, Lyons-Weiler MA, LaFramboise WA, Kelly L, Seethala RR, Grandis JR, Sobol RW and Taioli E: MicroRNA-137 promoter methylation is associated with poorer overall survival in patients with squamous cell carcinoma of the head and neck. Cancer. 117:1454–1462. 2011. View Article : Google Scholar : PubMed/NCBI
40	Liu LL, Lu SX, Li M, Li LZ, Fu J, Hu W, Yang YZ, Luo RZ, Zhang CZ and Yun JP: FoxD3-regulated microRNA-137 suppresses tumour growth and metastasis in human hepatocellular carcinoma by targeting AKT2. Oncotarget. 5:5113–5124. 2014. View Article : Google Scholar : PubMed/NCBI
41	Han F, Wang S, Chang Y, Li C, Yang J, Han Z, Chang B, Sun B and Chen L: Triptolide prevents extracellular matrix accumulation in experimental diabetic kidney disease by targeting microRNA-137/Notch1 pathway. J Cell Physiol. 233:2225–2237. 2018. View Article : Google Scholar
42	Tang CH, Chiu YC, Tan TW, Yang RS and Fu WM: Adiponectin enhances IL-6 production in human synovial fibroblast via an AdipoR1 receptor, AMPK, 38 and NF-kappa B pathway. J Immunol. 179:5483–5492. 2007. View Article : Google Scholar : PubMed/NCBI
43	Lihn AS, Jessen N, Pedersen SB, Lund S and Richelsen B: AICAR stimulates adiponectin and inhibits cytokines in adipose tissue. Biochem Biophys Res Commun. 316:853–858. 2004. View Article : Google Scholar : PubMed/NCBI
44	Paula FM, Leite NC, Vanzela EC, Kurauti MA, Freitas-Dias R, Carneiro EM, Boschero AC and Zoppi CC: Exercise increases pancreatic β-cell viability in a model of type 1 diabetes through IL-6 signaling. FASEB J. 29:1805–1816. 2015. View Article : Google Scholar : PubMed/NCBI
45	Almuraikhy S, Kafienah W, Bashah M, Diboun I, Jaganjac M, Al-Khelaifi F, Abdesselem H, Mazloum NA, Alsayrafi M, Mohamed-Ali V and Elrayess MA: Interleukin-6 induces impairment in human subcutaneous adipogenesis in obesity-associated insulin resistance. Diabetologia. 59:2406–2416. 2016. View Article : Google Scholar : PubMed/NCBI
46	Qu D, Liu J, Lau CW and Huang Y: IL-6 in diabetes and cardiovascular complications. Br J Pharmacol. 171:3595–3603. 2014. View Article : Google Scholar : PubMed/NCBI