Archives of Toxicology

, Volume 93, Issue 11, pp 3099–3109 | Cite as

Arsenic is more potent than cadmium or manganese in disrupting the INS-1 beta cell microRNA landscape

  • Rowan Beck
  • Mohit Chandi
  • Matt Kanke
  • Miroslav Stýblo
  • Praveen SethupathyEmail author
Inorganic Compounds


Diabetes is a metabolic disorder characterized by fasting hyperglycemia and impaired glucose tolerance. Laboratory and population studies have shown that inorganic arsenic (iAs) can impair these pathways. Other metals including cadmium (Cd) and manganese (Mn) have also been linked to diabetes phenotypes. MicroRNAs, short non-coding RNAs that regulate gene expression, have emerged as potential drivers of metabolic dysfunction. MicroRNAs responsive to metal exposures in vitro have also been reported in independent studies to regulate insulin secretion in vivo. We hypothesize that microRNA dysregulation may associate with and possibly contribute to insulin secretion impairment upon exposure to iAs, Cd, or Mn. We exposed insulin secreting rat insulinoma cells to non-cytotoxic concentrations of iAs (1 µM), Cd (5 µM), and Mn (25 µM) for 24 h followed by small RNA sequencing to identify dysregulated microRNAs. RNA sequencing was then performed to further investigate changes in gene expression caused by iAs exposure. While all three metals significantly inhibited glucose-stimulated insulin secretion, high-throughput sequencing revealed distinct microRNA profiles specific to each exposure. One of the most significantly upregulated microRNAs post-iAs treatment is miR-146a (~ + 2-fold), which is known to be activated by nuclear factor κB (NF-κB) signaling. Accordingly, we found by RNA-seq analysis that genes upregulated by iAs exposure are enriched in the NF-κB signaling pathway and genes down-regulated by iAs exposure are enriched in miR-146a binding sites and are involved in regulating beta cell function. Notably, iAs exposure caused a significant decrease in the expression of Camk2a, a calcium-dependent protein kinase that regulates insulin secretion, has been implicated in type 2 diabetes, and is a likely target of miR-146a. Further studies are needed to elucidate potential interactions among NF-kB, miR-146a, and Camk2a in the context of iAs exposure.


miRNA Arsenic Diabetes RNA Cadmium Manganese 



This work was funded by NIH (Grant Number R01DK105965), NIEHS (Grant Number R01ES022697), UNC Superfund by NIEHS (Grant Number 3P42ES005948-22S2), and NIGMS (Grant Number 5T32 GM007092).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

204_2019_2574_MOESM1_ESM.docx (131 kb)
Supplementary material 1 (DOCX 131 kb)


  1. Abiola M, Favier M, Christodoulou-Vafeiadou E et al (2009) Activation of Wnt/β-catenin signaling increases insulin sensitivity through a reciprocal regulation of Wnt10b and SREBP-1c in skeletal muscle cells. PLoS One 4:e8509. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Alipoor B, Ghaedi H, Meshkani R et al (2017) Association of MiR-146a expression and type 2 diabetes mellitus: a meta-analysis. Int J Mol Cell Med 6:156–163. CrossRefPubMedPubMedCentralGoogle Scholar
  3. Bagge A, Clausen TR, Larsen S et al (2012) MicroRNA-29a is up-regulated in beta-cells by glucose and decreases glucose-stimulated insulin secretion. Biochem Biophys Res Commun 426:266–272. CrossRefPubMedGoogle Scholar
  4. Baly DL, Curry DL, Keen CL, Hurley LS (1984) Effect of manganese deficiency on insulin secretion and carbohydrate homeostasis in rats. J Nutr 114:1438–1446. CrossRefPubMedGoogle Scholar
  5. Beck RF, Bommarito P, Douillet C et al (2018) Circulating miRNAs associated with arsenic exposure. Environ Sci Technol. CrossRefPubMedGoogle Scholar
  6. Belgardt B, Ahmed K, Spranger M et al (2015) The microRNA-200 family regulates pancreatic beta cell survival in type 2 diabetes. Nat Med 21:619–627. CrossRefPubMedGoogle Scholar
  7. Bhatt K, Lanting LL, Jia Y et al (2016) Anti-inflammatory role of microRNA-146a in the pathogenesis of diabetic nephropathy. J Am Soc Nephrol 27:2277–2288. CrossRefPubMedGoogle Scholar
  8. Bloomston M, Frankel WL, Petrocca F et al (2007) MicroRNA expression patterns to differentiate pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis. JAMA 297:1901. CrossRefPubMedGoogle Scholar
  9. Bouwens L, Rooman I (2005) Regulation of pancreatic beta-cell mass. Physiol Rev 85:1255–1270. CrossRefPubMedGoogle Scholar
  10. Chang K-C, Hsu C-C, Liu S-H et al (2013) Cadmium induces apoptosis in pancreatic β-cells through a mitochondria-dependent pathway: the role of oxidative stress-mediated c-Jun N-terminal kinase activation. PLoS One 8:e54374. CrossRefPubMedPubMedCentralGoogle Scholar
  11. Chen EY, Xu H, Gordonov S et al (2012) Expression2Kinases: mRNA profiling linked to multiple upstream regulatory layers. Bioinformatics 28:105–111. CrossRefPubMedGoogle Scholar
  12. Chen S, Feng B, Thomas AA, Chakrabarti S (2017) miR-146a regulates glucose induced upregulation of inflammatory cytokines extracellular matrix proteins in the retina and kidney in diabetes. PLoS One 12:e0173918. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Chi SW, Zang JB, Mele A, Darnell RB (2009) Argonaute HITS-CLIP decodes microRNA? mRNA interaction maps. Nature 460:479. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Dadi PK, Vierra NC, Ustione A et al (2014) Inhibition of pancreatic β-Cell Ca2+/calmodulin-dependent protein kinase II reduces glucose-stimulated calcium influx and insulin secretion, impairing glucose tolerance. J Biol Chem 289:12435–12445. CrossRefPubMedPubMedCentralGoogle Scholar
  15. Del Razo LM, García-Vargas GG, Valenzuela OL et al (2011) Exposure to arsenic in drinking water is associated with increased prevalence of diabetes: a cross-sectional study in the Zimapán and Lagunera regions in Mexico. Environ Health 10:73. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Díaz-Villaseñor A, Burns AL, Salazar AM et al (2008) Arsenite reduces insulin secretion in rat pancreatic β-cells by decreasing the calcium-dependent calpain-10 proteolysis of SNAP-25. Toxicol Appl Pharmacol 231:291–299. CrossRefPubMedGoogle Scholar
  17. Dixit SS, Wang T, Manzano EJQ et al (2013) Effects of CaMKII-mediated phosphorylation of ryanodine receptor type 2 on islet calcium handling, insulin secretion, and glucose tolerance. PLoS One 8:e58655. CrossRefPubMedPubMedCentralGoogle Scholar
  18. Douillet C, Currier J, Saunders J et al (2013) Methylated trivalent arsenicals are potent inhibitors of glucose stimulated insulin secretion by murine pancreatic islets. Toxicol Appl Pharmacol 267:11–15. CrossRefPubMedGoogle Scholar
  19. Douillet C, Huang MC, Saunders RJ et al (2017) Knockout of arsenic (+3 oxidation state) methyltransferase is associated with adverse metabolic phenotype in mice: the role of sex and arsenic exposure. Arch Toxicol 91:2617–2627. CrossRefPubMedGoogle Scholar
  20. Dover EN, Beck R, Huang MC et al (2017) Arsenite and methylarsonite inhibit mitochondrial metabolism and glucose-stimulated insulin secretion in INS-1 832/13 β cells. Arch Toxicol. CrossRefPubMedPubMedCentralGoogle Scholar
  21. Dover EN, Patel NY, Stýblo M (2018) Impact of in vitro heavy metal exposure on pancreatic β-cell function. Toxicol Lett 299:137–144. CrossRefPubMedGoogle Scholar
  22. Ediger BN, Du A, Liu J et al (2014) Islet-1 Is essential for pancreatic β-cell function. Diabetes 63:4206–4217. CrossRefPubMedPubMedCentralGoogle Scholar
  23. El Muayed M, Raja MR, Zhang X et al (2012) Accumulation of cadmium in insulin-producing β cells. Islets 4:405–416. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Fabricio G, Malta A, Chango A, De Freitas Mathias PC (2016) Environmental contaminants and pancreatic beta-cells. J Clin Res Pediatr Endocrinol 8:257–263. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Fang L, Cai J, Chen B et al (2015) Aberrantly expressed miR-582-3p maintains lung cancer stem cell-like traits by activating Wnt/β-catenin signalling. Nat Commun 6:8640. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Gribble MO, Howard BV, Umans JG et al (2012) Arsenic exposure, diabetes prevalence, and diabetes control in the Strong Heart Study. Am J Epidemiol 176:865–874. CrossRefPubMedPubMedCentralGoogle Scholar
  27. Hectors TLM, Vanparys C, van der Ven K et al (2011) Environmental pollutants and type 2 diabetes: a review of mechanisms that can disrupt beta cell function. Diabetologia 54:1273–1290. CrossRefPubMedGoogle Scholar
  28. Hohmeier HE, Mulder H, Chen G et al (2000) Isolation of INS-1-derived cell lines with robust ATP-sensitive K+ channel-dependent and-independent glucose-stimulated insulin secretion. Diabetes 49:424–430. CrossRefPubMedGoogle Scholar
  29. Huang MC, Douillet C, Su M et al (2016) Metabolomic profiles of arsenic (+3 oxidation state) methyltransferase knockout mice: effect of sex and arsenic exposure. Arch Toxicol. CrossRefPubMedPubMedCentralGoogle Scholar
  30. Huang MC, Douillet C, Dover EN, Stýblo M (2018) Prenatal arsenic exposure and dietary folate and methylcobalamin supplementation alter the metabolic phenotype of C57BL/6J mice in a sex-specific manner. Arch Toxicol 92:1925–1937. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Huang M, Douillet C, Stýblo M (2019) Arsenite and its trivalent methylated metabolites inhibit glucose-stimulated calcium influx and insulin secretion in murine pancreatic islets. Arch Toxicol. CrossRefPubMedGoogle Scholar
  32. IDF (2017) Diabetes atlas, 8th edn. International Diabetes Federation, BrusselsGoogle Scholar
  33. Jo S, Chen J, Xu G et al (2018) miR-204 controls glucagon-like peptide 1 receptor expression and agonist function. Diabetes 67:256–264. CrossRefPubMedGoogle Scholar
  34. Kitamura YI, Kitamura T, Kruse J-P et al (2005) FoxO1 protects against pancreatic β cell failure through NeuroD and MafA induction. Cell Metab 2:153–163. CrossRefPubMedGoogle Scholar
  35. Korc M (1983) Manganese action on pancreatic protein synthesis in normal and diabetic rats. Am J Physiol Liver Physiol 245:G628–G634. CrossRefGoogle Scholar
  36. Kuo C-C, Howard BV, Umans JG et al (2015) Arsenic exposure, arsenic metabolism, and incident diabetes in the Strong Heart Study. Diabetes Care. CrossRefPubMedPubMedCentralGoogle Scholar
  37. Lachmann A, Xu H, Krishnan J et al (2010) ChEA: transcription factor regulation inferred from integrating genome-wide ChIP-X experiments. Bioinformatics 26:2438–2444. CrossRefPubMedPubMedCentralGoogle Scholar
  38. Leung AKL, Young AG, Bhutkar A et al (2011) Genome-wide identification of Ago2 binding sites from mouse embryonic stem cells with and without mature microRNAs. Nat Struct Mol Biol 18:237–244. CrossRefPubMedPubMedCentralGoogle Scholar
  39. Li XT, Yu PF, Gao Y et al (2017) Association between plasma metal levels and diabetes risk: a case-control study in China. Biomed Environ Sci 30:482–491. CrossRefPubMedGoogle Scholar
  40. Liu S, Guo X, Wu B et al (2014) Arsenic induces diabetic effects through beta-cell dysfunction and increased gluconeogenesis in mice. Sci Rep 4:6894. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Liu B, Feng W, Wang J et al (2016) Association of urinary metals levels with type 2 diabetes risk in coke oven workers. Environ Pollut 210:1–8. CrossRefPubMedGoogle Scholar
  42. Lorenz MA, El Azzouny MA, Kennedy RT, Burant CF (2013) Metabolome response to glucose in the β-cell line INS-1 832/13. J Biol Chem 288:10923–10935. CrossRefPubMedPubMedCentralGoogle Scholar
  43. Makaji E, Raha S, Wade MG, Holloway AC (2011) Effect of environmental contaminants on beta cell function. Int J Toxicol 30:410–418. CrossRefPubMedGoogle Scholar
  44. Maull EA, Ahsan H, Edwards J et al (2012) Evaluation of the association between arsenic and diabetes: a National Toxicology Program workshop review. Environ Health Perspect 120:1658–1670. CrossRefPubMedPubMedCentralGoogle Scholar
  45. Mendez MA, González-Horta C, Sánchez-Ramírez B et al (2016) Chronic exposure to arsenic and markers of cardiometabolic risk: a cross-sectional study in Chihuahua, Mexico. Environ Health Perspect 124:104–111. CrossRefPubMedGoogle Scholar
  46. Ogata H, Goto S, Fujibuchi W, Kanehisa M (1998) Computation with the KEGG pathway database. Biosystems 47:119–128CrossRefGoogle Scholar
  47. Park K-G, Lee K-M, Seo H-Y et al (2007) Glucotoxicity in the INS-1 rat insulinoma cell line is mediated by the orphan nuclear receptor small heterodimer partner. Diabetes 56:431–437CrossRefGoogle Scholar
  48. Paul DS, Devesa V, Hernandez-Zavala A et al (2008) Environmental arsenic as a disruptor of insulin signaling. Met Ions Biol Med 10:1–7PubMedPubMedCentralGoogle Scholar
  49. Poy MN, Eliasson L, Krutzfeldt J et al (2004) A pancreatic islet-specific microRNA regulates insulin secretion. Nature 432:226–230. CrossRefPubMedGoogle Scholar
  50. Rhee SY, Hwang Y-C, Woo J et al (2013) Arsenic exposure and prevalence of diabetes mellitus in Korean adults. J Korean Med Sci 28:861. CrossRefPubMedPubMedCentralGoogle Scholar
  51. Rodríguez-Comas J, Moreno-Asso A, Moreno-Vedia J et al (2017) Stress-induced MicroRNA-708 impairs β-cell function and growth. Diabetes 66:3029–3040. CrossRefPubMedGoogle Scholar
  52. Roggli E, Britan A, Gattesco S et al (2010) Involvement of microRNAs in the cytotoxic effects exerted by proinflammatory cytokines on pancreatic β-cells. Diabetes 59:978–986. CrossRefPubMedPubMedCentralGoogle Scholar
  53. Ruepp A, Brauner B, Dunger-Kaltenbach I et al (2008) CORUM: the comprehensive resource of mammalian protein complexes. Nucleic Acids Res 36:D646–D650. CrossRefPubMedGoogle Scholar
  54. Santos GJD, Ferreira SM, Ortis F et al (2014) Metabolic memory of β-cells controls insulin secretion and is mediated by CaMKII. Mol Metab 3:484–489. CrossRefPubMedPubMedCentralGoogle Scholar
  55. Schug J, McKenna LB, Walton G et al (2013) Dynamic recruitment of microRNAs to their mRNA targets in the regenerating liver. BMC Genom 14:264. CrossRefGoogle Scholar
  56. Schwartz GG, Il’yasova D, Ivanova A (2003) Urinary cadmium, impaired fasting glucose, and diabetes in the NHANES III. Diabetes Care 26:468–470. CrossRefPubMedGoogle Scholar
  57. Shan Z, Chen S, Sun T et al (2016) U-shaped association between plasma manganese levels and type 2 diabetes. Environ Health Perspect 124:1876–1881. CrossRefPubMedPubMedCentralGoogle Scholar
  58. Skelin M, Rupnik M, Cencic A (2010) Pancreatic beta cell lines and their applications in diabetes mellitus research. Altex 27:105–113 (doi:20686743) CrossRefGoogle Scholar
  59. Sung T-C, Huang J-W, Guo H-R (2015) Association between arsenic exposure and diabetes: a meta-analysis. Biomed Res Int 2015:368087. CrossRefPubMedPubMedCentralGoogle Scholar
  60. Taganov KD, Boldin MP, Chang K-J, Baltimore D (2006) NF-kappaB-dependent induction of microRNA miR-146, an inhibitor targeted to signaling proteins of innate immune responses. Proc Natl Acad Sci USA 103:12481–12486. CrossRefPubMedGoogle Scholar
  61. Trajkovski M, Hausser J, Soutschek J et al (2011) MicroRNAs 103 and 107 regulate insulin sensitivity. Nature 474:649–653. CrossRefPubMedGoogle Scholar
  62. Treviño S, Waalkes MP, Flores Hernández JA et al (2015) Chronic cadmium exposure in rats produces pancreatic impairment and insulin resistance in multiple peripheral tissues. Arch Biochem Biophys 583:27–35. CrossRefPubMedGoogle Scholar
  63. Vrijens K, Bollati V, Nawrot TS (2015) MicroRNAs as potential signatures of environmental exposure or effect: a systematic review. Environ Health Perspect. CrossRefPubMedPubMedCentralGoogle Scholar
  64. Waldsee R, Eftekhari S, Ahnstedt H et al (2014) CaMKII and MEK1/2 inhibition time-dependently modify inflammatory signaling in rat cerebral arteries during organ culture. J Neuroinflamm 11:90. CrossRefGoogle Scholar
  65. Wang W, Xie Z, Lin Y, Zhang D (2014) Association of inorganic arsenic exposure with type 2 diabetes mellitus: a meta-analysis. J Epidemiol Community Health 68:176–184. CrossRefPubMedGoogle Scholar
  66. Wang H, Ren Y, Hu X et al (2017) Effect of Wnt signaling on the differentiation of Islet β-cells from adipose-derived stem cells. Biomed Res Int. CrossRefPubMedPubMedCentralGoogle Scholar
  67. Yajima H, Komatsu M, Schermerhorn T et al (1999) cAMP enhances insulin secretion by an action on the ATP-sensitive K+ channel-independent pathway of glucose signaling in rat pancreatic islets. Diabetes 48:1006–1012. CrossRefPubMedGoogle Scholar
  68. Yang W, Wang J, Chen Z et al (2017) NFE2 induces miR-423-5p to promote gluconeogenesis and hyperglycemia by repressing the hepatic FAM3A-ATP-Akt pathway. Diabetes 66:1819–1832. CrossRefPubMedGoogle Scholar
  69. Zhang P (2017) CaMKII: the molecular villain that aggravates cardiovascular disease. Exp Ther Med 13:815–820. CrossRefPubMedPubMedCentralGoogle Scholar
  70. Zhong P, Huang H (2017) CaMKII as a pathological mediator of inflammation, oxidative stress, ER stress, autophagy and mitochondrial dysfunction in free fatty acids/hyperlipidemia-induced cardiac remodeling both in vitro and vivo. J Mol Cell Cardiol 112:134. CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Genetics, School of MedicineUniversity of North Carolina at Chapel HillChapel HillUSA
  2. 2.Department of Biomedical Sciences, College of Veterinary MedicineCornell UniversityIthacaUSA
  3. 3.Department of Nutrition, Gillings School of Global Public HealthUniversity of North Carolina at Chapel HillChapel HillUSA

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