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Proliferation of human lens epithelial cells (HLE-B3) is inhibited by blocking of voltage-gated calcium channels

  • Cell and Molecular Physiology
  • Published:
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Abstract

Calcium, as an integral part of a large number of cellular regulatory pathways, is selective in the control of specific cell functions like the start of G1 phase in cell cycle. Cell proliferation has been suggested to depend on increasing intracellular calcium levels. A major regulatory pathway for intracellular calcium is the calcium influx into the cell via voltage-gated calcium channels. T-type and L-type calcium channels are substantially present in human lens epithelial cell (hLEC), and total calcium currents are inhibited by mibefradil. Here, the hypothesis was tested if calcium influx via Cav channels regulates proliferation in epithelial cells. Cell proliferation was determined by cell culture assays using the L- and T-type Cav channel blockers mibefradil and verapamil as modulators for calcium influx. Calcium influx was investigated using the Manganese quench technique. Western blot experiments were accomplished under standard conditions using antibodies against MAPK 3. Mibefradil as well as verapamil impaired cell proliferation, but in different concentration ranges. Furthermore, the activation of MAPK 3 was reduced by both antagonists. Calcium influx was also reduced in the presence of both blockers. We conclude that the transmembrane influx of Ca2+ through Cav channels contributes to the regulation of hLEC proliferation, identifying Cav channel blockers as potential therapeutic substances in ocular diseases.

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References

  1. Bertelmann E, Kojetinsky C (2001) Posterior capsule opacification and anterior capsule opacification. Curr Opin Ophthalmol 12:35–40

    Article  PubMed  CAS  Google Scholar 

  2. Lu L, Reinach PS, Kao WW (2001) Corneal epithelial wound healing. Exp Biol Med (Maywood) 226:653–664

    CAS  Google Scholar 

  3. Whitfield JF, Bird RP, Chakravarthy BR, Isaacs RJ, Morley P (1995) Calcium-cell cycle regulator, differentiator, killer, chemopreventor, and maybe, tumor promoter. J Cell Biochem Suppl 22:74–91

    Article  PubMed  CAS  Google Scholar 

  4. Silver RB (1999) Imaging structured space-time patterns of Ca2+ signals: essential information for decisions in cell division. FASEB J 13(Suppl 2):S209–S215

    PubMed  CAS  Google Scholar 

  5. Akaike N, Brown AM, Nishi K, Tsuda Y (1981) Actions of verapamil, diltiazem and other divalent cations on the calcium-current of helix neurones. Br J Pharmacol 74:87–95

    PubMed  CAS  Google Scholar 

  6. McRory JE, Santi CM, Hamming KS et al (2001) Molecular and functional characterization of a family of rat brain T-type calcium channels. J Biol Chem 276:3999–4011

    Article  PubMed  CAS  Google Scholar 

  7. Catterall WA (1995) Structure and function of voltage-gated ion channels. Annu Rev Biochem 64:493–531

    Article  PubMed  CAS  Google Scholar 

  8. Chemin J, Monteil A, Lory P (2003) [Functional specificity of T-type calcium channels and their roles in neuronal differentiation]. J Soc Biol 197:235–247

    PubMed  CAS  Google Scholar 

  9. Perez-Reyes E (1998) Molecular characterization of a novel family of low voltage-activated, T-type, calcium channels. J Bioenerg Biomembr 30:313–318

    Article  PubMed  CAS  Google Scholar 

  10. Kuga T, Kobayashi S, Hirakawa Y, Kanaide H, Takeshita A (1996) Cell cycle-dependent expression of L- and T-type Ca2+ currents in rat aortic smooth muscle cells in primary culture. Circ Res 79:14–19

    PubMed  CAS  Google Scholar 

  11. Lory P, Bidaud I, Chemin J (2006) T-type calcium channels in differentiation and proliferation. Cell Calcium 40:135–146

    Article  PubMed  CAS  Google Scholar 

  12. Panner A, Cribbs LL, Zainelli GM, Origitano TC, Singh S, Wurster RD (2005) Variation of T-type calcium channel protein expression affects cell division of cultured tumor cells. Cell Calcium 37:105–119

    Article  PubMed  CAS  Google Scholar 

  13. Nebe B, Kunz F, Peters A, Rychly J, Noack T, Beck R (2004) Induction of apoptosis by the calcium antagonist mibefradil correlates with depolarization of the membrane potential and decreased integrin expression in human lens epithelial cells. Graefes Arch Clin Exp Ophthalmol 242:597–604

    Article  PubMed  CAS  Google Scholar 

  14. Meissner A, Nebe B, Beck R, Guthoff R, Noack T (2007) Importance of calcium and potassium currents in human lens epithelial cells and the effect of the calcium channel blocker mibefradil. Trace Elem Electrolytes 24:228–234

    CAS  Google Scholar 

  15. Rampe D, Anderson B, Rapien-Pryor V, Li T, Dage RC (1993) Comparison of the in vitro and in vivo cardiovascular effects of two structurally distinct Ca++ channel activators, BAY K 8644 and FPL 64176. J Pharmacol Exp Ther 265:1125–1130

    PubMed  CAS  Google Scholar 

  16. Enslen H, Tokumitsu H, Stork PJ, Davis RJ, Soderling TR (1996) Regulation of mitogen-activated protein kinases by a calcium/calmodulin-dependent protein kinase cascade. Proc Natl Acad Sci U S A 93:10803–10808

    Article  PubMed  CAS  Google Scholar 

  17. Hook SS, Means AR (2001) Ca(2+)/CaM-dependent kinases: from activation to function. Annu Rev Pharmacol Toxicol 41:471–505

    Article  PubMed  CAS  Google Scholar 

  18. Illario M, Cavallo AL, Bayer KU et al (2003) Calcium/calmodulin-dependent protein kinase II binds to Raf-1 and modulates integrin-stimulated ERK activation. J Biol Chem 278:45101–45108

    Article  PubMed  CAS  Google Scholar 

  19. Gazzaniga P, Gradilone A, Vercillo R et al (1996) Bcl-2/bax mRNA expression ratio as prognostic factor in low-grade urinary bladder cancer. Int J Cancer 69:100–104

    Article  PubMed  CAS  Google Scholar 

  20. Curtis TM, Scholfield CN (2001) Nifedipine blocks Ca2+ store refilling through a pathway not involving L-type Ca2+ channels in rabbit arteriolar smooth muscle. J Physiol 532:609–623

    Article  PubMed  CAS  Google Scholar 

  21. Zhao PL, Wang XT, Zhang XM et al (2002) Tubular and cellular localization of the cardiac L-type calcium channel in rat kidney. Kidney Int 61:1393–1406

    Article  PubMed  CAS  Google Scholar 

  22. Clevenger CV, Epstein AL, Bauer KD (1987) Modulation of the nuclear antigen p105 as a function of cell-cycle progression. J Cell Physiol 130:336–343

    Article  PubMed  CAS  Google Scholar 

  23. Mariot P, Vanoverberghe K, Lalevee N, Rossier MF, Prevarskaya N (2002) Overexpression of an alpha 1H (Cav3.2) T-type calcium channel during neuroendocrine differentiation of human prostate cancer cells. J Biol Chem 277:10824–10833

    Article  PubMed  CAS  Google Scholar 

  24. McCobb DP, Best PM, Beam KG (1989) Development alters the expression of calcium currents in chick limb motoneurons. Neuron 2:1633–1643

    Article  PubMed  CAS  Google Scholar 

  25. Clozel JP, Ertel EA, Ertel SI (1997) Discovery and main pharmacological properties of mibefradil (Ro 40-5967), the first selective T-type calcium channel blocker. J Hypertens Suppl 15:S17–S25

    PubMed  CAS  Google Scholar 

  26. Bezprozvanny I, Tsien RW (1995) Voltage-dependent blockade of diverse types of voltage-gated Ca2+ channels expressed in Xenopus oocytes by the Ca2+ channel antagonist mibefradil (Ro 40-5967). Mol Pharmacol 48:540–549

    PubMed  CAS  Google Scholar 

  27. Chouabe C, Drici MD, Romey G, Barhanin J, Lazdunski M (1998) HERG and KvLQT1/IsK, the cardiac K+ channels involved in long QT syndromes, are targets for calcium channel blockers. Mol Pharmacol 54:695–703

    PubMed  CAS  Google Scholar 

  28. Gomora JC, Enyeart JA, Enyeart JJ (1999) Mibefradil potently blocks ATP-activated K(+) channels in adrenal cells. Mol Pharmacol 56:1192–1197

    PubMed  CAS  Google Scholar 

  29. McNulty MM, Kyle JW, Lipkind GM, Hanck DA (2006) An inner pore residue (Asn406) in the Nav1.5 channel controls slow inactivation and enhances mibefradil block to T-type Ca2+ channel levels. Mol Pharmacol 70:1514–1523

    Article  PubMed  CAS  Google Scholar 

  30. Nilius B, Prenen J, Kamouchi M, Viana F, Voets T, Droogmans G (1997) Inhibition by mibefradil, a novel calcium channel antagonist, of Ca(2+)- and volume-activated Cl− channels in macrovascular endothelial cells. Br J Pharmacol 121:547–555

    Article  PubMed  CAS  Google Scholar 

  31. Perchenet L, Clement-Chomienne O (2000) Characterization of mibefradil block of the human heart delayed rectifier hKv1.5. J Pharmacol Exp Ther 295:771–778

    PubMed  CAS  Google Scholar 

  32. Strege PR, Bernard CE, Ou Y, Gibbons SJ, Farrugia G (2005) Effect of mibefradil on sodium and calcium currents. Am J Physiol Gastrointest Liver Physiol 289:G249–G253

    Article  PubMed  CAS  Google Scholar 

  33. Chemin J, Nargeot J, Lory P (2004) Ca(v)3.2 calcium channels control an autocrine mechanism that promotes neuroblastoma cell differentiation. Neuroreport 15:671–675

    Article  PubMed  CAS  Google Scholar 

  34. Gray LS, Perez-Reyes E, Gomora JC et al (2004) The role of voltage gated T-type Ca2+ channel isoforms in mediating “capacitative” Ca2+ entry in cancer cells. Cell Calcium 36:489–497

    Article  PubMed  CAS  Google Scholar 

  35. Bertolesi GE, Jollimore CA, Shi C et al (2003) Regulation of alpha1G T-type calcium channel gene (CACNA1G) expression during neuronal differentiation. Eur J Neurosci 17:1802–1810

    Article  PubMed  Google Scholar 

  36. Rodman DM, Reese K, Harral J et al (2005) Low-voltage-activated (T-type) calcium channels control proliferation of human pulmonary artery myocytes. Circ Res 96:864–872

    Article  PubMed  CAS  Google Scholar 

  37. Panner A, Wurster RD (2006) T-type calcium channels and tumor proliferation. Cell Calcium 40:253–259

    Article  PubMed  CAS  Google Scholar 

  38. Schreiber R (2005) Ca2+ signaling, intracellular pH and cell volume in cell proliferation. J Membr Biol 205:129–137

    Article  PubMed  CAS  Google Scholar 

  39. Bijlenga P, Liu JH, Espinos E et al (2000) T-type alpha 1H Ca2+ channels are involved in Ca2+ signaling during terminal differentiation (fusion) of human myoblasts. Proc Natl Acad Sci U S A 97:7627–7632

    Article  PubMed  CAS  Google Scholar 

  40. Choi J, Park SY, Joo CK (2004) Hepatocyte growth factor induces proliferation of lens epithelial cells through activation of ERK1/2 and JNK/SAPK. Invest Ophthalmol Vis Sci 45:2696–2704

    Article  PubMed  Google Scholar 

  41. Dai G, Zhang Z, Zhou M, Yu S (2002) [The role of mitogen-activated protein kinase (MAPK) pathway in the action mechanism of PYM-induced KB cells apoptosis]. Zhonghua Kou Qiang Yi Xue Za Zhi 37:97–98

    PubMed  CAS  Google Scholar 

  42. Rampe D, Janis RA, Triggle DJ (1984) BAY K 8644, a 1,4-dihydropyridine Ca2+ channel activator: dissociation of binding and functional effects in brain synaptosomes. J Neurochem 43:1688–1692

    Article  PubMed  CAS  Google Scholar 

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  1. Institute of physiology, University of Rostock, Gertrudenstrasse 9, Rostock, 18055, Germany

    Anja Meissner & Thomas Noack

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  1. Anja Meissner
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  2. Thomas Noack
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Correspondence to Anja Meissner.

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Meissner, A., Noack, T. Proliferation of human lens epithelial cells (HLE-B3) is inhibited by blocking of voltage-gated calcium channels. Pflugers Arch - Eur J Physiol 457, 47–59 (2008). https://doi.org/10.1007/s00424-008-0514-5

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  • DOI: https://doi.org/10.1007/s00424-008-0514-5

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