Skip to main content
SpringerLink
Log in

Cell cycle-dependent subcellular distribution of ClC-3 in HeLa cells

  • Original Paper
  • Published:
Histochemistry and Cell Biology Aims and scope Submit manuscript

Abstract

Chloride channel-3 (ClC-3) is suggested to be a component and/or a regulator of the volume-activated Cl− channel in the plasma membrane. However, ClC-3 is predominantly located inside cells and the role of intracellular ClC-3 in tumor growth is unknown. In this study, we found that the subcellular distribution of endogenous ClC-3 varied in a cell cycle-dependent manner in HeLa cells. During interphase, ClC-3 was distributed throughout the cell and it accumulated at various positions in different stages. In early G1, ClC-3 was mainly located in the nucleus. In middle G1, ClC-3 gathered around the nuclear periphery as a ring. In late G1, ClC-3 moved back into the nucleus, where it remained throughout S phase. In G2, ClC-3 was concentrated in the cytoplasm. When cells progressed from G2 to the prophase of mitosis, ClC-3 from the cytoplasm translocated into the nucleus. During metaphase and anaphase, ClC-3 was distributed throughout the cell except for around the chromosomes and was aggregated at the spindle poles and in between two chromosomes, respectively. ClC-3 was then again concentrated in the nucleus upon the progression from telophase to cytokinesis. These results reveal a cell cycle-dependent change of the subcellular distribution of ClC-3 and strongly suggest that ClC-3 has nucleocytoplasmic shuttling dynamics that may play key regulatory roles during different stages of the cell cycle in tumor cells.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

References

  • Aldehni F, Spitzner M, Martins JR, Barro-Soria R, Schreiber R, Kunzelmann K (2009) Bestrophin 1 promotes epithelial-to-mesenchymal transition of renal collecting duct cells. J Am Soc Nephrol 20:1556–1564

    Article  PubMed  CAS  Google Scholar 

  • Aoto T, Saitoh N, Sakamoto Y, Watanabe S, Nakao M (2008) Polycomb group protein-associated chromatin is reproduced in post-mitotic G1 phase and is required for S phase progression. J Biol Chem 283:18905–18915

    Article  PubMed  CAS  Google Scholar 

  • Bauerschmidt C, Pollok S, Kremmer E, Nasheuer HP, Grosse F (2007) Interactions of human Cdc45 with the Mcm2-7 complex, the GINS complex, and DNA polymerases delta and epsilon during S phase. Genes Cells 12:745–758

    PubMed  CAS  Google Scholar 

  • Bonaccorsi S, Giansanti MG, Gatti M (2000) Spindle assembly in Drosophila neuroblasts and ganglion mother cells. Nat Cell Biol 2:54–56

    Article  PubMed  CAS  Google Scholar 

  • Chen L, Wang L, Zhu L, Nie S, Zhang J, Zhong P, Cai B, Luo H, Jacob TJ (2002) Cell cycle-dependent expression of volume-activated chloride currents in nasopharyngeal carcinoma cells. Am J Physiol Cell Physiol 283:C1313–C1323

    PubMed  CAS  Google Scholar 

  • Chen LX, Zhu LY, Jacob TJ, Wang LW (2007) Roles of volume-activated Cl- currents and regulatory volume decrease in the cell cycle and proliferation in nasopharyngeal carcinoma cells. Cell Prolif 40:253–267

    Article  PubMed  CAS  Google Scholar 

  • Chen Y, Sanchez A, Rubio ME, Kohl T, Pardo LA, Stuhmer W (2011) Functional K(v)10.1 channels localize to the inner nuclear membrane. PLoS ONE 6:e19257

    Article  PubMed  CAS  Google Scholar 

  • Do CW, Lu W, Mitchell CH, Civan MM (2005) Inhibition of swelling-activated Cl- currents by functional anti-ClC-3 antibody in native bovine non-pigmented ciliary epithelial cells. Invest Ophthalmol Vis Sci 46:948–955

    Article  PubMed  Google Scholar 

  • Doyle AGJ, Newell DG (1993) Cell and tissue culture: laboratory procedures. Wiley, Chichester

    Google Scholar 

  • Duan DD (2011) The ClC-3 chloride channels in cardiovascular disease. Acta Pharmacol Sin 32:675–684

    Article  PubMed  CAS  Google Scholar 

  • Duan D, Zhong J, Hermoso M, Satterwhite CM, Rossow CF, Hatton WJ, Yamboliev I, Horowitz B, Hume JR (2001) Functional inhibition of native volume-sensitive outwardly rectifying anion channels in muscle cells and Xenopus oocytes by anti-ClC-3 antibody. J Physiol 531:437–444

    Article  PubMed  CAS  Google Scholar 

  • Gaffney EV (1975) The accumulation and selective detachment of mitotic cells. Methods Cell Biol 9:71–84

    Article  PubMed  CAS  Google Scholar 

  • Gentzsch M, Cui L, Mengos A, Chang XB, Chen JH, Riordan JR (2003) The PDZ-binding chloride channel ClC-3B localizes to the golgi and associates with cystic fibrosis transmembrane conductance regulator-interacting PDZ proteins. J Biol Chem 278:6440–6449

    Article  PubMed  CAS  Google Scholar 

  • Habela CW, Olsen ML, Sontheimer H (2008) ClC3 is a critical regulator of the cell cycle in normal and malignant glial cells. J Neurosci 28:9205–9217

    Article  PubMed  CAS  Google Scholar 

  • Hara-Chikuma M, Yang B, Sonawane ND, Sasaki S, Uchida S, Verkman AS (2005) ClC-3 chloride channels facilitate endosomal acidification and chloride accumulation. J Biol Chem 280:1241–1247

    Article  PubMed  CAS  Google Scholar 

  • Hermoso M, Satterwhite CM, Andrade YN, Hidalgo J, Wilson SM, Horowitz B, Hume JR (2002) ClC-3 is a fundamental molecular component of volume-sensitive outwardly rectifying Cl- channels and volume regulation in HeLa cells and Xenopus laevis oocytes. J Biol Chem 277:40066–40074

    Article  PubMed  CAS  Google Scholar 

  • Jentsch TJ, Maritzen T, Keating DJ, Zdebik AA, Thevenod F (2010) ClC-3—a granular anion transporter involved in insulin secretion? Cell Metab 12:307–308 (author reply 309–310)

    Article  PubMed  CAS  Google Scholar 

  • Jin NG, Kim JK, Yang DK, Cho SJ, Kim JM, Koh EJ, Jung HC, So I, Kim KW (2003) Fundamental role of ClC-3 in volume-sensitive Cl- channel function and cell volume regulation in AGS cells. Am J Physiol Gastrointest Liver Physiol 285:G938–G948

    PubMed  CAS  Google Scholar 

  • Kakino S, Sasaki K, Kurose A, Ito H (1996) Intracellular localization of cyclin B1 during the cell cycle in glioma cells. Cytometry 24:49–54

    Article  PubMed  CAS  Google Scholar 

  • Li W (2008) Expression and clinical significance of CLC-3 mRNA in cervical cancer. China Medical University Graduate School, vol Master. China Medical University, Shenyang, p 36

    Google Scholar 

  • Li X, Wang T, Zhao Z, Weinman SA (2002) The ClC-3 chloride channel promotes acidification of lysosomes in CHO-K1 and Huh-7 cells. Am J Physiol Cell Physiol 282:C1483–C1491

    PubMed  CAS  Google Scholar 

  • Majoul I, Schmidt T, Pomasanova M, Boutkevich E, Kozlov Y, Soling HD (2002) Differential expression of receptors for Shiga and Cholera toxin is regulated by the cell cycle. J Cell Sci 115:817–826

    PubMed  CAS  Google Scholar 

  • Mao J, Chen L, Xu B, Wang L, Wang W, Li M, Zheng M, Li H, Guo J, Li W, Jacob TJ (2009) Volume-activated chloride channels contribute to cell-cycle-dependent regulation of HeLa cell migration. Biochem Pharmacol 77:159–168

    Article  PubMed  CAS  Google Scholar 

  • Mao J, Xu B, Li H, Chen L, Jin X, Zhu J, Wang W, Zhu L, Zuo W, Chen W, Wang L (2011) Lack of association between stretch-activated and volume-activated Cl currents in hepatocellular carcinoma cells. J Cell Physiol 226:1176–1185

    Article  PubMed  CAS  Google Scholar 

  • Matsuda JJ, Filali MS, Volk KA, Collins MM, Moreland JG, Lamb FS (2008) Overexpression of CLC-3 in HEK293T cells yields novel currents that are pH dependent. Am J Physiol Cell Physiol 294:C251–C262

    Article  PubMed  CAS  Google Scholar 

  • Nigg EA (2001) Mitotic kinases as regulators of cell division and its checkpoints. Nat Rev Mol Cell Biol 2:21–32

    Article  PubMed  CAS  Google Scholar 

  • Pines J, Hunter T (1991) Human cyclins A and B1 are differentially located in the cell and undergo cell cycle-dependent nuclear transport. J Cell Biol 115:1–17

    Article  PubMed  CAS  Google Scholar 

  • Remillard CV, Yuan JX (2005) ClC-3: more than just a volume-sensitive Cl- channel. Br J Pharmacol 145:1–2

    Article  PubMed  CAS  Google Scholar 

  • Shen MR, Droogmans G, Eggermont J, Voets T, Ellory JC, Nilius B (2000) Differential expression of volume-regulated anion channels during cell cycle progression of human cervical cancer cells. J Physiol 529(Pt 2):385–394

    Article  PubMed  CAS  Google Scholar 

  • Shimada K, Li X, Xu G, Nowak DE, Showalter LA, Weinman SA (2000) Expression and canalicular localization of two isoforms of the ClC-3 chloride channel from rat hepatocytes. Am J Physiol Gastrointest Liver Physiol 279:G268–G276

    PubMed  CAS  Google Scholar 

  • Sontheimer H (2008) An unexpected role for ion channels in brain tumor metastasis. Exp Biol Med (Maywood) 233:779–791

    Article  CAS  Google Scholar 

  • Tang YB, Liu YJ, Zhou JG, Wang GL, Qiu QY, Guan YY (2008) Silence of ClC-3 chloride channel inhibits cell proliferation and the cell cycle via G/S phase arrest in rat basilar arterial smooth muscle cells. Cell Prolif 41:775–785

    Article  PubMed  CAS  Google Scholar 

  • Wang L, Chen L, Zhu L, Rawle M, Nie S, Zhang J, Ping Z, Kangrong C, Jacob TJ (2002) Regulatory volume decrease is actively modulated during the cell cycle. J Cell Physiol 193:110–119

    Article  PubMed  CAS  Google Scholar 

  • Wang LW, Chen LX, Jacob T (2004) ClC-3 expression in the cell cycle of nasopharyngeal carcinoma cells. Sheng Li Xue Bao 56:230–236

    PubMed  CAS  Google Scholar 

  • Wang B, Song L, Wu C (2007) Expression and distribution of ClC-3 in human pulmonary carcinoma. J Fourth Mil Med Univ 28:1509–1511

    CAS  Google Scholar 

  • Xu B, Luo Q, Wang X, Liu X, Wang Y, Sun L (2005) Expression and distribution of C1C–3 in human glioma specimen. Chinese J Pathophysiol 21:2188–2191

    CAS  Google Scholar 

  • Xu B, Mao J, Wang L, Zhu L, Li H, Wang W, Jin X, Zhu J, Chen L (2010) ClC-3 chloride channels are essential for cell proliferation and cell cycle progression in nasopharyngeal carcinoma cells. Acta Biochim Biophys Sin (Shanghai) 42:370–380

    Article  CAS  Google Scholar 

  • Zhang HN, Zhou JG, Qiu QY, Ren JL, Guan YY (2006) ClC-3 chloride channel prevents apoptosis induced by thapsigargin in PC12 cells. Apoptosis 11:327–336

    Article  PubMed  CAS  Google Scholar 

  • Zhao W, Wang Y, Han Z, Sun L, Zhang L (2009) Expression of the CLC-3 in the breast cancer and its clinical significances. Clin J Lab Diagn 13:350–352

    Google Scholar 

Download references

Acknowledgments

This work was supported by the National Natural Science Foundation of China (30800435, 81170339, 81101666, 30771106, 30871267 and 30870567).

Author information

Authors and Affiliations

  1. Department of Biology and Guangdong Provincial Key Laboratory of Pharmaceutical Bioactive Substances, Guangzhou Higher Education Mega Center, Guangdong Pharmaceutical University, Guangzhou, 510006, China

    Jianwen Mao, Xiaobo Li, Weiqiang Chen, Bin Xu, Hongzhi Li, Xiaobao Jin, Jiayong Zhu, Guixian lin & Weizhang Wang

  2. Department of Physiology, Medical College, Jinan University, Guangzhou, 510632, China

    Haifeng Zhang & Liwei Wang

  3. Department of Pharmacology, Medical College, Jinan University, Guangzhou, 510632, China

    Jianwen Mao & Lixin Chen

Authors
  1. Jianwen Mao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaobo Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Weiqiang Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bin Xu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Haifeng Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hongzhi Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Liwei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiaobao Jin
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jiayong Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Guixian lin
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Weizhang Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Lixin Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Hongzhi Li or Lixin Chen.

Additional information

J. Mao, X. Li and W. Chen contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mao, J., Li, X., Chen, W. et al. Cell cycle-dependent subcellular distribution of ClC-3 in HeLa cells. Histochem Cell Biol 137, 763–776 (2012). https://doi.org/10.1007/s00418-012-0937-0

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00418-012-0937-0

Keywords

Search

Navigation