Advertisement

Molecular and Cellular Biochemistry

, Volume 397, Issue 1–2, pp 305–312 | Cite as

Altered expression of genes associated with telomere maintenance and cell function of human vascular endothelial cell at elevated temperature

  • Toyoki Maeda
  • Jing-Zhi Guan
  • Masamichi Koyanagi
  • Naoki Makino
Article

Abstract

The pathophysiological alterations of vascular endothelial cells induced by heat were studied. Human umbilical venous endothelial cells were cultured for 1 day at three different temperatures (37, 39, and 42 °C). The telomere lengths, the expressions of proteins associated with telomere length maintenance, apoptosis, heat shock, and vascular function were analyzed. The cell growth was not suppressed at 39 °C but suppressed at 42 °C. The mean telomere length did not change, whereas the telomere length distribution altered at 42 °C. Long telomere decreased and middle-sized telomere increased in the telomere length distribution at 42 °C. The telomerase activity did not show any heat-associated alterations. However, of the components of telomerase, telomerase reverse transcriptase was up-regulated along temperature elevation. In contrast, the expression level of RNA component TERC did not altered. Among the analyzed apoptosis-associated proteins, p21 was down-regulated and phosphorylated p53 was up-regulated. Heat shock proteins and NO synthase were up-regulated at 42 °C. These results suggested that induced growth suppression or cell senescence was induced by strong heat stress rather than mild one predominantly in cells bearing long telomeres with p53 activation, and simultaneously activated some telomere-associated factors, heat shock proteins, and NO synthesis probably for heat-resistant cell survival.

Keywords

Heat stress Vascular endothelial cell Telomere Telomerase Apoptosis 

Abbreviations

HUVECs

Human umbilical venous endothelial cells

TERT

Telomerase reverse transcriptase

TERC

Telomerase RNA component

Hsp

Heat shock proteins

eNOS

Endothelial nitric oxide synthase

PD

Population doubling

SA-β-Gal

Senescence-associated β-galactosidase

TL

Telomere length

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

TRAP

Telomerase repeat amplification protocol

ANOVA

Analysis of variance

Notes

Acknowledgments

We would like to thank Ms. K. Tsuchida, Ms. S. Taguchi, and Ms. Y. Ueda for their expert technical assistance. This work was supported by Grants from the Ministry of Education, Science, and Culture of Japan (#23590885), the National Natural Science Fund (NSFC) (81170329/H2501), and 2012 Health and Labour Sciences Research Grants Comprehensive Research on Life Style Related Diseases including Cardiovascular Diseases and Diabetes Mellitus.

Conflict of interest

The authors have no financial competing interest to declare in relation to this manuscript.

References

  1. 1.
    Blackburn EH (1991) Structure and function of telomeres. Nature (London) 350:569–573CrossRefGoogle Scholar
  2. 2.
    Maeda T, Guan JZ, Oyama J, Higuchi Y, Makino N (2009) Aging-associated alteration in subtelomeric methylation in Parkinson’s disease. J Gerontol A Biol Sci Med Sci 64:949–955PubMedCrossRefGoogle Scholar
  3. 3.
    Aubert G, Lansdorp PM (2008) Telomeres and aging. Physiol Rev 88:557–579PubMedCrossRefGoogle Scholar
  4. 4.
    Njemini R, Bautmans I, Onyema OO, Van Puyvelde K, Demanet C, Mets T (2011) Circulating heat shock protein 70 in health, aging and disease. BMC Immunol 12:24PubMedCrossRefPubMedCentralGoogle Scholar
  5. 5.
    Xu Q, Metzler B, Jahangiri M, Mandal K (2012) Molecular chaperones and heat shock proteins in atherosclerosis. Am J Physiol Heart Circ Physiol 302:H506–H514PubMedCrossRefPubMedCentralGoogle Scholar
  6. 6.
    Kellogg DL Jr, Crandall CG, Liu Y, Charkoudian N, Johnson JM (1998) Nitric oxide and cutaneous active vasodilation during heat stress in humans. J Appl Physiol 85:824–829PubMedGoogle Scholar
  7. 7.
    Guan JZ, Maeda T, Sugano M, Oyama J, Higuchi Y, Makino N (2007) Change in the telomere length distribution with age in the Japanese population. Mol Cell Biochem 304:253–260CrossRefGoogle Scholar
  8. 8.
    Tao Q, Lv B, Qiao B, Zheng CQ, Chen ZF (2009) Immortalization of ameloblastoma cells via reactivation of telomerase function: phenotypic and molecular characteristics. Oral Oncol 45:e239–e244PubMedCrossRefGoogle Scholar
  9. 9.
    Fuiwara M, Kamma H, Wu W, Yano Y, Homma S, Satoh H (2006) Alternative lengthening of telomeres in the human adrenocortical carcinoma cell line H295R. Int J Oncol 29:445–451Google Scholar
  10. 10.
    Makino N, Maeda T, Oyama J-I, Higuchi Y, Mimori K (2009) Improving insulin sensitivity via activation of PPAR-γ increases telomerase activity in the heart of OLETF rats. Am J Physiol Heart Circ Physiol 297:H2188–H2195PubMedCrossRefGoogle Scholar
  11. 11.
    Sugano M, Tsuchida K, Maeda T, Makino N (2007) siRNA targeting SHP-1 accelerates angiogenesis in a rat model of hindlimb ischemia. Atherosclerosis 191:33–39PubMedCrossRefGoogle Scholar
  12. 12.
    Maeda T, Kurita R, Yokoo T, Tani K, Makino N (2011) Telomerase inhibition promotes an initial step of cell differentiation of primate embryonic stem cell. Biochem Biophys Res Commun 407:491–494PubMedCrossRefGoogle Scholar
  13. 13.
    Zakian VA (1995) Telomeres: beginning to understand the end. Science 270:1601–1607PubMedCrossRefGoogle Scholar
  14. 14.
    Leel J, Sunjg YH, Cheong C, Choi YS, Jeon HK, Sun W, Hahn WC, Ishikawa F, Lee HW (2008) TERT promotes cellular and organismal survival independently of telomerase activity. Oncogene 27:3754–3760CrossRefGoogle Scholar
  15. 15.
    Biroccio A, Rizzo A, Elli R, Koering CE, Belleville A, Benassi B, Leonetti C, Stevens MF, D’Incalci M, Zupi G, Gilson E (2006) TRF2 inhibition triggers apoptosis and reduces tumorigenicity of human melanoma cells. Eur J Cancer 42:1881–1888PubMedCrossRefGoogle Scholar
  16. 16.
    Smogorzewska A, de Lange T (2002) Different telomere damage signaling pathways in human and mouse cells. EMBO J 21:4338–4348PubMedCrossRefPubMedCentralGoogle Scholar
  17. 17.
    Zhang P, Dilley C, Mattson MP (2007) DNA damage responses in neural cells: focus on the telomere. Neuroscience 145:1439–1448PubMedCrossRefPubMedCentralGoogle Scholar
  18. 18.
    Cheng A, Shin-ya K, Wan R, Tang SC, Miura T, Tang H, Khatri R, Gleichman M, Ouyang X, Liu D, Park HR, Chiang JY, Mattson MP (2007) Telomere protection mechanisms change during neurogenesis and neuronal maturation: newly generated neurons are hypersensitive to telomere and DNA damage. J Neurosci 27:3722–3733PubMedCrossRefGoogle Scholar
  19. 19.
    Madan E, Gogna R, Pati U (2012) p53 Ser15 phosphorylation disrupts the p53-RPA70 complex and induces RPA70-mediated DNA repair in hypoxia. Biochem J 443:811–820PubMedCrossRefGoogle Scholar
  20. 20.
    Douville JM, Cheung DY, Herbert KL, Moffatt T, Wigle JT (2011) Mechanisms of MEOX1 and MEOX2 regulation of the cyclin dependent kinase inhibitors p21 and p16 in vascular endothelial cells. PLoS ONE 6:e29099PubMedCrossRefPubMedCentralGoogle Scholar
  21. 21.
    Chen YC, Lin-Shiau SY, Lin JKP (1999) Involvement of heat-shock protein 70 and P53 proteins in attenuation of UVC-induced apoptosis by thermal stress in hepatocellular carcinoma cells. Photochem Photobiol 70:78–86PubMedCrossRefGoogle Scholar
  22. 22.
    Matsumoto H, Shimura M, Omatsu T, Okaichi K, Majima H, Ohnishi T (1994) p53 proteins accumulated by heat stress associate with heat shock proteins HSP72/HSC73 in human glioblastoma cell lines. Cancer Lett 87:39–46PubMedCrossRefGoogle Scholar
  23. 23.
    Li PC, Yang CC, Hsu SP, Chien CT (2012) Repetitive progressive thermal preconditioning hinders thrombosis by reinforcing phosphatidylinositol 3-kinase/Akt-dependent heat-shock protein/endothelial nitric oxide synthase signaling. J Vasc Surg 56:159–170PubMedCrossRefGoogle Scholar
  24. 24.
    Bree RT, Stenson-Cox C, Grealy M, Byrnes L, Gorman AM, Samali A (2002) Cellular longevity: role of apoptosis and replicative senescence. Biogerontology 3:195–206PubMedCrossRefGoogle Scholar
  25. 25.
    Richardson PG, Mitsiades CS, Laubach JP, Lonial S, Chanan-Khan AA, Anderson KC (2011) Inhibition of heat shock protein 90 (HSP90) as a therapeutic strategy for the treatment of myeloma and other cancers. Br J Haematol 152:367–379PubMedCrossRefGoogle Scholar
  26. 26.
    Ota H, Eto M, Kano MR, Kahyo T, Setou M, Ogawa S, Iijima K, Akishita M, Ouchi Y (2010) Induction of endothelial nitric oxide synthase, by statins inhibits endothelial senescence through the Akt pathway. Arterioscler Thromb Vasc Biol 30:2205–2211PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Toyoki Maeda
    • 1
  • Jing-Zhi Guan
    • 2
  • Masamichi Koyanagi
    • 1
  • Naoki Makino
    • 1
  1. 1.The Department of Cardiovascular, Respiratory, and Geriatric MedicineKyushu University Beppu HospitalBeppuJapan
  2. 2.The 309th Hospital of Chinese People’s Liberation ArmyBeijingChina

Personalised recommendations