Gestational diabetes leads to down-regulation of CDK4-pRB-E2F1 pathway genes in pancreatic islets of rat offspring

Document Type : Original Article


1 Department of Animal Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran

2 Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran

3 Stem Cell Research Center, Faculty of Medicine, Golestan University of Medical Sciences, Gorgan, Iran

4 Gorgan Congenital Malformations Research Center, Depatment of Anatomical Sciences, Golestan University of Medical Sciences, Gorgan, Iran


Objective(s): The link between a hyperglycemic intrauterine environment and the development of diabetes later in life has been observed in offspring exposed to gestational diabetes mellitus (GDM), but the underlying mechanisms for this phenomenon are still not clear. Reduced β-cells mass is a determinant in the development of diabetes (type 1 and type 2 diabetes). Some recent studies have provided evidence that the CDK4-pRB-E2F1 regulatory pathway is involved in β-cells proliferation. Therefore, we postulated that GDM exposure impacts the offspring’s β-cells by disruption in the CDK4-pRB-E2F1 pathway.
Materials and Methods: Adult Wistar rats were randomly allocated in control and diabetic group. The experimental group received 40 mg/kg/body weight of streptozotocin (STZ) on day zero of gestation. After delivery, diabetic offspring of GDM mothers and control dams at the age of 15 week were randomly scarified and pancreases were harvested. Langerhans islets of diabetic and control groups were digested by collagenase digestion technique. After RNA extraction, we investigated the expressions of the kir 6.2 and CDK4-pRB-E2F1 pathway genes by quantitative real-time PCR.
Results: GDM reduced the expression of CDK4-pRB-E2F1 pathway genes in Langerhans islets cells of offspring. CDK4, pRB and E2F1 pathway genes were downregulated in diabetic islets by 51%, 35% and 84%, respectively. Also, the expression of Kir 6.2 was significantly decreased in diabetic islets by 88%.
Conclusion: We suggest that the effect of gestational diabetes on offspring’s β-cells may be primarily caused by the suppression of CDK4-pRB-E2F1 pathway.


1.Golalipour MJ, Ghafari S, Farsi M. Effect of urtica dioica L extract on quantitative morphometric alterations of liver parenchymal cells in STZ diabetic rats. Int J Morphol 2009; 27:1339-1344.
2. American Diabetes Association (ADA). Standards of medical care in diabetes. Diabetes Care 2014; 37:14–80.
3. Mitanchez D, Yzydorczy Ck, Siddeek B, Boubred F, Benahmed M, Simeoni U. The offspring of the diabetic mother – Short- and long-term implications. Best Pract Res Clin Obstet Gynaecol 2015; 29:256-269.
4. Cohen MM, Shiota K. Teratogenesis of holopro-sencephaly. Am J Med Genet 2002; 109:1–15.
5. Correa A, Gilboa SM, Besser LM, Botto LD, Moore CA, Hobbs CA, et al. Diabetes mellitus and birth defects. Am J Obstet Gynecol 2008; 199:237–237.
6. Simmons RA, Templeton LJ, Gertz SJ. Intrauterine growth retardation leads to the development of type 2 diabetes in the rat. Diabetes 2001; 50:2279–2286.
7. Boloker J, Gertz SJ, Simmons RA. Gestational diabetes leads to the development of diabetes in adulthood in the rat. Diabetes 2002; 51:1499-1506.
8. Min Y, Lowy C, Ghebremeskel K, Thomas B, Offley-Shore B, Crawford M. Unfavourable effect of type 1 and type 2 diabetes on maternal and fetal essential fatty acid status: a potential marker of fetal insulin resistance. Am J Clin Nutr 2005; 82:1162–1168.
9. Tata VD. Age-related impairmen tof pancreatic beta-cell unction:pathophysiological and cellular mechanisms. Front Endocrinol 2014; 138:1-8.
10. Bruning JC, Winnay J, Cheatham B, Kahn CR. Differential signaling by insulin receptor substrate 1 (IRS-1) and IRS-2 in IRS-1-deficient cells. Mol Cell Biol 1997; 17:1513-1521.
11. Butler AE, Janson J, Bonner-Weir S, Ritzel R, Rizza RA, Butler PC. Beta-cell deficit and increased beta-cell apoptosis in humans with type 2 diabetes. Diabetes 2003; 52:102–110.
12. Meier JJ, Bhushan A, Butler AE, Rizza RA, Butler PC. Sustained beta cell apoptosis in patients with long-standing type 1 diabetes: indirect evidence for islet regeneration? Diabetologia 2005; 48:2221–2228.
13. Rane SG, Dubus P, Mettus RV, Galbreath EJ, Boden G, Reddy EP, et al. Loss of Cdk4 expression causes insulin-deficient diabetes and Cdk4 activation results in beta-islet cell hyperplasia. Nat Genet 1999; 22:44–52.
14. Martin J, Hunt SL, Dubus P, Sotillo R, Néhmé-Pélluard F, Magnuson MA, et al. Genetic rescue of Cdk4 null mice restores pancreatic beta-cell proliferation but not homeostatic cell number. Oncogene 2003; 22:5261–5269.
15. Dalle S, Sardet C, Fajas L. The CDK4-pRB-E2F1 pathway controls insulin secretion. Nat Cell Biol 2009; 11:1017-1023.
16. Fajas L, Blanchet E, Annicotte JS. CDK4, pRB and E2F1: connected to insulin. Cell Division 2010; 5:6.
17. Pasek, RC, Gannon M. Advancements and challenges in generating accurate animal models of gestational diabetes mellitus. Am J Physiol Endocrinol Metab 2013; 305:1327–1338.
18. Carter JD, Dula SB, Corbin KL, Wu R, Nunemaker CS. Practical guide to rodent islet isolation and assessment. Biol Proced Online 2009; 11:3-31.
19. Matveyenko AV, Veldhuis JD, Butler PC. Adaptations in pulsatile insulin secretion, hepatic insulin clearance, and beta cell mass to age-related insulin resistance in rats. Am J Physiol Endocrinol Metab 2008; 295:832–841.
20. Ahangarpour A, Teymuri H, Jabari A, Malekshahi Nia H, Heidari H. Antidiabetic and hypolipidemic effects of Dorema aucheri hydroalcoholic leave extract in streptozotocin-nicotinamide induced type 2 diabetes in male rats. Iran J Basic Med Sci 2014; 17:808-814.
 21. Georgua S, Bhushan A. Beta cell replication is the primary mechanism for maintaining postnatal beta cell mass. J Clin Invest 2004; 114:963-968.
22. Kushner JA, Ciemerych MA, Sicinska E, Wartschow LM, Teta M, Long SY, et al. Cyclins D2 and D1 are essential for postnatal pancreatic beta-cell growth. Mol Cell Biol 2005; 25:3752–3762.
23. Teta M, Long SY, Wartschow LM, Rankin MM, Kushner JA. Very slow turnover of β-cells in aged adult mice. Diabetes 2005; 54:2557–2567.
24. Meier JJ, Butler AE, Saisho Y, Monchamp T, Galasso R, Bhushan A, et al. Beta-cell replication is the primary mechanism subserving the postnatal expansion of beta-cell mass in humans. Diabetes 2008; 57:1584–1594.
25. Aerts L, Holemans K, Van AF. Maternal diabetes during pregnancy: consequences for the offspring. Diabetes Metab 1990; 6:147–167.
26. Rieck S, White P, Schug J, Fox AJ, Smirnova O, Gao N. The transcriptional response of the islet to pregnancy in mice. Mol Endocrinol 2009; 23:1702–1712.
27. Fajas L, Annicotte JS, Miard S, Sarruf D, Watanabe M, Auwerx J. Impaired pancreatic growth, beta cell mass, and beta cell function in E2F1-/- mice. J Clin Invest 2004; 113:1288-1295.