Evaluation of protein levels of autophagy markers (Beclin 1 and SQSTM1/p62) and phosphorylation of cyclin E in the placenta of women with preeclampsia

Dilara Akcora Yildiz; Sevgi Irtegun Kandemir; Elif Agacayak; Engin Deveci

doi:10.14715/10.14715/cmb/2017.63.12.12

Issue

Vol. 63 No. 12: Issue 12

Issue Published : December 19, 2017

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Evaluation of protein levels of autophagy markers (Beclin 1 and SQSTM1/p62) and phosphorylation of cyclin E in the placenta of women with preeclampsia

https://doi.org/10.14715/10.14715/cmb/2017.63.12.12

Dilara Akcora Yildiz

Department of Biology, Faculty of Science and Arts, Mehmet Akif Ersoy University, Burdur, Turkey

Sevgi Irtegun Kandemir

Department of Medical Biology, Faculty of Medicine, Dicle University, Diyarbakir, Turkey

Elif Agacayak

Department of Gynecology and Obstetrics, Faculty of Medicine, Dicle University, Diyarbakir, Turkey

Engin Deveci

4 Department of Histology, Faculty of Medicine, Dicle University, Diyarbakir, Turkey

Corresponding Author(s) : Dilara Akcora Yildiz

dilaraakcora@mehmetakif.edu.tr

Cellular and Molecular Biology, Vol. 63 No. 12: Issue 12
Article Published : December 30, 2017

Abstract

Preeclampsia is a severe multisystem disorder, and its pathophysiology is still not completely understood. Autophagy, a recycling process that maintains cellular homoeostasis during differentiation and development, is controversial regarding increased or decreased autophagic activity in preeclampsia. The aim of this study was to determine whether autophagy is increased in the placentas of women with preeclampsia by examining the protein levels of autophagy markers (Beclin 1 and SQSTM1/p62) and phosphorylation of cyclin E. For this purpose, placentas from preeclampsia (n=10) and control (n=10) pregnancies were included in this study. The protein expression of autophagy-related markers Beclin1, SQSTM1/p62 and phosphorylation status of cyclin E were detected by Western blot. Our data showed that the protein levels of both Beclin 1 and SQSTM1/p62 were significantly increased, while the phosphorylation level of cyclin E was significantly decreased in placentas with preeclampsia compared to those derived from controls. The results of this study suggest that the autophagic activity is perpetually increased in preeclampsia and cyclin E protein stabilisation might be involved in the induction of autophagy.

Keywords

Preeclampsia Placenta Beclin 1 SQSTM1/p62 Cyclin E.

Akcora Yildiz, D., Irtegun Kandemir, S., Agacayak, E., & Deveci, E. (2017). Evaluation of protein levels of autophagy markers (Beclin 1 and SQSTM1/p62) and phosphorylation of cyclin E in the placenta of women with preeclampsia. Cellular and Molecular Biology, 63(12), 51–55. https://doi.org/10.14715/10.14715/cmb/2017.63.12.12

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References

Hutcheon J. A., Lisonkova S., Joseph K.S. Epidemiology of pre-eclampsia and the other hypertensive disorders of pregnancy. Best Pract Res Clin Obstet Gynaecol. 2011; 25: 391-403.
Trogstad L., Magnus P., Stoltenberg C. Pre-eclampsia: Risk factors and causal models. Best Pract Res Clin Obstet Gynaecol. 2011; 25: 329-342.
Velicky P, Knofler M, Pollheimer J. Function and control of human invasive trophoblast subtypes: Intrinsic vs. maternal control. Cell Adh Migr. 2016; 10: 154-162.
James J. L., Whitley G. S., Cartwright J. E. Pre-eclampsia: fitting together the placental, immune and cardiovascular pieces. J Pathol. 2010; 221: 363–378.
He C., Klionsky D.J., Regulation Mechanisms and Signaling Pathways of Autophagy. Annu Rev Genet. 2009; 43: 67–93.
Cecconi F, Levine B. The role of autophagy in mammalian development: cell makeover rather than cell death. Dev Cell. 2008;15: 344-57.
Mizushima N, Ohsumi Y, Yoshimori T. Autophagosome formation in mammalian cells. Cell Struct Funct. 2002; 27(6):421-9.
Yorimitsu T, Klionsky DJ. Autophagy: molecular machinery for self-eating. Cell Death Differ. 2005;12 Suppl 2:1542-52.
Kang R, Zeh HJ, Lotze MT, Tang D. The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ. 2011;18(4):571-80.
Seibenhener ML, Babu JR, Geetha T, Wong HC, Krishna NR, Wooten MW. Sequestosome 1/p62 is a polyubiquitin chain binding protein involved in ubiquitin proteasome degradation. Mol Cell Biol. 2004;24(18):8055-68.
Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, et al. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem. 2007;282(33):24131-45.
Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000;19(21):5720-8.
Katsuragi Y, Ichimura Y, Komatsu M. p62/SQSTM1 functions as a signaling hub and an autophagy adaptor. FEBS J. 2015 Dec;282(24):4672-8.
Kanninen TT, de Andrade Ramos BR, Witkin SS. The role of autophagy in reproduction from gametogenesis to parturition. Eur J Obstet Gynecol Reprod Biol. 2013;171(1):3-8.
Chifenti B, Locci MT, Lazzeri G, Guagnozzi M, Dinucci D, Chiellini F, et al. Autophagy-related protein LC3 and Beclin-1 in the first trimester of pregnancy. Clin Exp Reprod Med. 2013;40(1):33-7.
Avagliano L, Terraneo L, Virgili E, Martinelli C, Doi P, Samaja M, et al. Autophagy in Normal and Abnormal Early Human Pregnancies. Reprod Sci. 2015;22(7):838-44.
Signorelli P, Avagliano L, Virgili E, Gagliostro V, Doi P, Braidotti P, et al. Autophagy in term normal human placentas. Placenta. 2011;32(6):482-5.
Curtis S, Jones CJ, Garrod A, Hulme CH, Heazell AE. Identification of autophagic vacuoles and regulators of autophagy in villous trophoblast from normal term pregnancies and in fetal growth restriction. J Matern Fetal Neonatal Med. 2013;26(4):339-46.
Hwang HC, Clurman BE. Cyclin E in normal and neoplastic cell cycles. Oncogene. 2005;24(17):2776-86.
Parisi T, Beck AR, Rougier N, McNeil T, Lucian L, Werb Z, Amati B. Cyclins E1 and E2 are required for endoreplication in placental trophoblast giant cells. EMBO J. 2003;22(18):4794-803.
Irtegun S, Tekin MA, Alpaycı R. Increased Expression of E-cadherin, Endothelin-1, and CD68 in Preeclamptic Placentas. Erciyes Med J. 2016; 38(4): 149-52.
Mull BB, Cox J, Bui T, Keyomarsi K. Post-translational modification and stability of low molecular weight cyclin E. Oncogene. 2009; 28: 3167–3176.
Ugland H, Naderi S, Brech A, Collas P, Blomhoff HK. cAMP induces autophagy via a novel pathway involving ERK, cyclin E and Beclin 1. Autophagy. 2011;7(10):1199-211.
Oh SY, Choi SJ, Kim KH, Cho EY, Kim JH, Roh CR. Autophagy-related proteins, LC3 and Beclin-1, in placentas from pregnancies complicated by preeclampsia. Reprod Sci. 2008 Nov;15(9):912-20.
Gao L, Qi HB, Kamana KC, Zhang XM, Zhang H, Baker PN. Excessive autophagy induces the failure of trophoblast invasion and vasculature: possible relevance to the pathogenesis of preeclampsia. J Hypreeclampsiartens. 2015 Jan;33(1):106-17.
Akaishi R, Yamada T, Nakabayashi K, Nishihara H, Furuta I, Kojima T, et al. Autophagy in the placenta of women with hypertensive disorders in pregnancy. Placenta. 2014 Dec;35(12):974-80.
Hung TH, Chen SF, Lo LM, Li MJ, Yeh YL, Hsieh TT. et al. PLoS One. 2012;7(7):e40957.
Zhang Y, Hu X, Gao G, Wang Y, Chen P, Ye Y. Autophagy protects against oxidized low density lipoprotein-mediated inflammation associated with preeclampsia. Placenta. 2016 Dec;48:136-143.
Nakashima A, Aoki A, Kusabiraki T, Shima T, Yoshino O, Cheng SB, et al. Role of autophagy in oocytogenesis, embryogenesis, implantation, and pathophysiology of pre-eclampsia. J Obstet Gynaecol Res. 2017 Apr;43(4):633-643.
Olvera M, Harris S, Amezcua CA, McCourty A, Rezk S, Koo C, et al. Immunohistochemical expression of cell cycle proteins E2F-1, Cdk-2, Cyclin E, p27(kip1), and Ki-67 in normal placenta and gestational trophoblastic disease. Mod Pathol. 2001 Oct;14(10):1036-42.
Mull BB, Cox J, Bui T, Keyomarsi K. Post-translational modification and stability of low molecular weight cyclin E. Oncogene. 2009 Sep 3;28(35):3167-76.
Gao Q, Zhu X, Chen J, Mao C, Zhang L, Xu Z. Upregulation of P53 promoted G1 arrest and apoptosis in human umbilical cord vein endothelial cells from preeclampsia. J Hypertens. 2016 Jul;34(7):1380-8.

References

Hutcheon J. A., Lisonkova S., Joseph K.S. Epidemiology of pre-eclampsia and the other hypertensive disorders of pregnancy. Best Pract Res Clin Obstet Gynaecol. 2011; 25: 391-403.

Trogstad L., Magnus P., Stoltenberg C. Pre-eclampsia: Risk factors and causal models. Best Pract Res Clin Obstet Gynaecol. 2011; 25: 329-342.

Velicky P, Knofler M, Pollheimer J. Function and control of human invasive trophoblast subtypes: Intrinsic vs. maternal control. Cell Adh Migr. 2016; 10: 154-162.

James J. L., Whitley G. S., Cartwright J. E. Pre-eclampsia: fitting together the placental, immune and cardiovascular pieces. J Pathol. 2010; 221: 363–378.

He C., Klionsky D.J., Regulation Mechanisms and Signaling Pathways of Autophagy. Annu Rev Genet. 2009; 43: 67–93.

Cecconi F, Levine B. The role of autophagy in mammalian development: cell makeover rather than cell death. Dev Cell. 2008;15: 344-57.

Mizushima N, Ohsumi Y, Yoshimori T. Autophagosome formation in mammalian cells. Cell Struct Funct. 2002; 27(6):421-9.

Yorimitsu T, Klionsky DJ. Autophagy: molecular machinery for self-eating. Cell Death Differ. 2005;12 Suppl 2:1542-52.

Kang R, Zeh HJ, Lotze MT, Tang D. The Beclin 1 network regulates autophagy and apoptosis. Cell Death Differ. 2011;18(4):571-80.

Seibenhener ML, Babu JR, Geetha T, Wong HC, Krishna NR, Wooten MW. Sequestosome 1/p62 is a polyubiquitin chain binding protein involved in ubiquitin proteasome degradation. Mol Cell Biol. 2004;24(18):8055-68.

Pankiv S, Clausen TH, Lamark T, Brech A, Bruun JA, Outzen H, et al. p62/SQSTM1 binds directly to Atg8/LC3 to facilitate degradation of ubiquitinated protein aggregates by autophagy. J Biol Chem. 2007;282(33):24131-45.

Kabeya Y, Mizushima N, Ueno T, Yamamoto A, Kirisako T, Noda T, et al. LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J. 2000;19(21):5720-8.

Katsuragi Y, Ichimura Y, Komatsu M. p62/SQSTM1 functions as a signaling hub and an autophagy adaptor. FEBS J. 2015 Dec;282(24):4672-8.

Kanninen TT, de Andrade Ramos BR, Witkin SS. The role of autophagy in reproduction from gametogenesis to parturition. Eur J Obstet Gynecol Reprod Biol. 2013;171(1):3-8.

Chifenti B, Locci MT, Lazzeri G, Guagnozzi M, Dinucci D, Chiellini F, et al. Autophagy-related protein LC3 and Beclin-1 in the first trimester of pregnancy. Clin Exp Reprod Med. 2013;40(1):33-7.

Avagliano L, Terraneo L, Virgili E, Martinelli C, Doi P, Samaja M, et al. Autophagy in Normal and Abnormal Early Human Pregnancies. Reprod Sci. 2015;22(7):838-44.

Signorelli P, Avagliano L, Virgili E, Gagliostro V, Doi P, Braidotti P, et al. Autophagy in term normal human placentas. Placenta. 2011;32(6):482-5.

Curtis S, Jones CJ, Garrod A, Hulme CH, Heazell AE. Identification of autophagic vacuoles and regulators of autophagy in villous trophoblast from normal term pregnancies and in fetal growth restriction. J Matern Fetal Neonatal Med. 2013;26(4):339-46.

Hwang HC, Clurman BE. Cyclin E in normal and neoplastic cell cycles. Oncogene. 2005;24(17):2776-86.

Parisi T, Beck AR, Rougier N, McNeil T, Lucian L, Werb Z, Amati B. Cyclins E1 and E2 are required for endoreplication in placental trophoblast giant cells. EMBO J. 2003;22(18):4794-803.

Irtegun S, Tekin MA, Alpaycı R. Increased Expression of E-cadherin, Endothelin-1, and CD68 in Preeclamptic Placentas. Erciyes Med J. 2016; 38(4): 149-52.

Mull BB, Cox J, Bui T, Keyomarsi K. Post-translational modification and stability of low molecular weight cyclin E. Oncogene. 2009; 28: 3167–3176.

Ugland H, Naderi S, Brech A, Collas P, Blomhoff HK. cAMP induces autophagy via a novel pathway involving ERK, cyclin E and Beclin 1. Autophagy. 2011;7(10):1199-211.

Oh SY, Choi SJ, Kim KH, Cho EY, Kim JH, Roh CR. Autophagy-related proteins, LC3 and Beclin-1, in placentas from pregnancies complicated by preeclampsia. Reprod Sci. 2008 Nov;15(9):912-20.

Gao L, Qi HB, Kamana KC, Zhang XM, Zhang H, Baker PN. Excessive autophagy induces the failure of trophoblast invasion and vasculature: possible relevance to the pathogenesis of preeclampsia. J Hypreeclampsiartens. 2015 Jan;33(1):106-17.

Akaishi R, Yamada T, Nakabayashi K, Nishihara H, Furuta I, Kojima T, et al. Autophagy in the placenta of women with hypertensive disorders in pregnancy. Placenta. 2014 Dec;35(12):974-80.

Hung TH, Chen SF, Lo LM, Li MJ, Yeh YL, Hsieh TT. et al. PLoS One. 2012;7(7):e40957.

Zhang Y, Hu X, Gao G, Wang Y, Chen P, Ye Y. Autophagy protects against oxidized low density lipoprotein-mediated inflammation associated with preeclampsia. Placenta. 2016 Dec;48:136-143.

Nakashima A, Aoki A, Kusabiraki T, Shima T, Yoshino O, Cheng SB, et al. Role of autophagy in oocytogenesis, embryogenesis, implantation, and pathophysiology of pre-eclampsia. J Obstet Gynaecol Res. 2017 Apr;43(4):633-643.

Olvera M, Harris S, Amezcua CA, McCourty A, Rezk S, Koo C, et al. Immunohistochemical expression of cell cycle proteins E2F-1, Cdk-2, Cyclin E, p27(kip1), and Ki-67 in normal placenta and gestational trophoblastic disease. Mod Pathol. 2001 Oct;14(10):1036-42.

Mull BB, Cox J, Bui T, Keyomarsi K. Post-translational modification and stability of low molecular weight cyclin E. Oncogene. 2009 Sep 3;28(35):3167-76.

Gao Q, Zhu X, Chen J, Mao C, Zhang L, Xu Z. Upregulation of P53 promoted G1 arrest and apoptosis in human umbilical cord vein endothelial cells from preeclampsia. J Hypertens. 2016 Jul;34(7):1380-8.

Author biographies is not available.