Home LiteratureArticle Details
PMID: 9763633 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Involvement of PKC-alpha in regulatory volume decrease responses and activation of volume-sensitive chloride channels in human cervical cancer HT-3 cells.

The Journal of physiology ·Vol. 512 ( Pt 2) ·1998-10-15 ·Pages 435-48

Chou CY, Shen MR, Hsu KS, Huang HY, Lin HC

Abstract

1. The present study was carried out to identify the specific protein kinase C (PKC) isoform involved in regulatory volume decrease (RVD) responses, and to investigate the signal transduction pathways underlying the activation of volume-sensitive chloride channels in human cervical cancer HT-3 cells. The role of Ca2+ in RVD and in the activation of chloride currents was also studied. 2. The time course of RVDs was prolonged by microinjection of PKC-alpha antibody but not by PKC-beta or PKC-gamma antibody, and also by exposure to Ca2+-free medium, in particular when combined with microinjection of EDTA. Immunofluorescence staining showed that hypotonic superfusion evoked the translocation of PKC-alpha to the cell membrane, whereas PKC-beta or PKC-gamma remained unaffected. The translocation of PKC-alpha was observed a few minutes after hypotonic stress, reaching peak intensity at 30 min, and returned to the cytoplasm 60 min after hypotonic exposure. Western blot analyses showed an increased PKC-alpha level in terms of intensity and phosphorylation in the cell membrane, while neither PKC-beta nor PKC-gamma was activated upon hyposmotic challenge. 3. Whole-cell patch-clamp studies demonstrated that neomycin and PKC blockers such as staurosporine and H7 inhibited volume-sensitive chloride currents. The inhibitory effect of neomycin on chloride currents can be reversed by the PKC activator phorbol 12-myristate, 13-acetate (PMA). Moreover, the PKC inhibitor and PKC-alpha antibody, but not PKC-beta or PKC-gamma antibody, significantly attenuated the chloride currents. The activation of volume-sensitive chloride currents were insensitive to the changes of intracellular Ca2+ but required the presence of extracellular Ca2+. 4. Our results suggest the involvement of PKC-alpha and extracellular Ca2+ in RVD responses and the activation of volume-sensitive chloride channels in HT-3 cells.

MeSH Terms
Blotting, Western Calcium Signaling/physiology Cell Size Chloride Channels/physiology Electric Stimulation Electrophysiology Female Fluorescent Antibody Technique, Direct Humans Isoenzymes/physiology Membrane Potentials/physiology Patch-Clamp Techniques Protein Kinase C/physiology Protein Kinase C-alpha Signal Transduction/physiology Subcellular Fractions/physiology Tumor Cells, Cultured Uterine Cervical Neoplasms/pathology
Chemicals
Chloride Channels Isoenzymes PRKCA protein, human Protein Kinase C Protein Kinase C-alpha
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Chou C Y
Department of Obstetrics and Gynecology, National Cheng Kung University Medical College, Tainan 704, Taiwan. chougyn@mail.ncku.edu.tw
Shen M R
Hsu K S
Huang H Y
Lin H C
References (48)
48 references, click to expand
  1. Volume-activated taurine transport is differentially activated in human cervical cancer HT-3 cells but not in human papillomavirus-immortalized Z183A and normal cervical epithelial cells.
    Clin Exp Pharmacol Physiol. 1997 Dec;24(12):935-9 PMID: 9406659
  2. Cl- and K+ conductances activated by cell swelling in primary cultures of rabbit distal bright convoluted tubules.
    Am J Physiol. 1997 Nov;273(5 Pt 2):F680-97 PMID: 9374831
  3. Differential osmosensing signalling pathways and G-protein involvement in human cervical cells with different tumour potential.
    Cell Signal. 1998 Feb;10(2):113-20 PMID: 9481486
  4. Microinjection of tissue culture cells.
    Methods Enzymol. 1983;101:482-92 PMID: 6310338
  5. Autophosphorylation of rat brain Ca2+-activated and phospholipid-dependent protein kinase.
    J Biol Chem. 1986 Sep 15;261(26):12134-40 PMID: 3462187
  6. Domain structure and phosphorylation of protein kinase C.
    J Biol Chem. 1987 Feb 15;262(5):2291-7 PMID: 3818597
  7. A thousand and one protein kinases.
    Cell. 1987 Sep 11;50(6):823-9 PMID: 3113737
  8. Autophosphorylation of the protein kinase dependent on double-stranded RNA.
    J Biol Chem. 1987 Nov 15;262(32):15538-44 PMID: 3479429
  9. Membrane mechanisms in volume and pH regulation in vertebrate cells.
    Physiol Rev. 1989 Apr;69(2):315-82 PMID: 2538851
  10. Biosynthesis and posttranslational modifications of protein kinase C in human breast cancer cells.
    J Biol Chem. 1989 Aug 15;264(23):13902-9 PMID: 2474538
  11. A point mutation at the putative ATP-binding site of protein kinase C alpha abolishes the kinase activity and renders it down-regulation-insensitive. A molecular link between autophosphorylation and down-regulation.
    J Biol Chem. 1990 Apr 15;265(11):6296-300 PMID: 2318854
  12. Hypotonicity stimulates phosphatidylcholine hydrolysis and generates diacylglycerol in erythrocytes.
    J Biol Chem. 1990 Aug 5;265(22):13055-9 PMID: 2376588
  13. Down-regulation of a kinase defective PKC-alpha.
    FEBS Lett. 1991 Jun 17;284(1):120-2 PMID: 2060619
  14. Signalling targets for anticancer drug development.
    Trends Pharmacol Sci. 1991 May;12(5):188-94 PMID: 1862534
  15. Epidermal growth factor enhances a microsomal 12-lipoxygenase activity in A431 cells.
    J Biol Chem. 1992 Feb 25;267(6):3657-66 PMID: 1740418
  16. Studies on the phosphorylation of protein kinase C-alpha.
    Biochem J. 1992 Apr 15;283 ( Pt 2):515-8 PMID: 1575696
  17. Reversal of the malignant phenotype by an anti-ras ribozyme.
    Antisense Res Dev. 1992 Spring;2(1):3-15 PMID: 1422085
  18. Calcium signaling in cell volume regulation.
    Physiol Rev. 1992 Oct;72(4):1037-61 PMID: 1332089
  19. Overexpression of protein kinase C-delta and -epsilon in NIH 3T3 cells induces opposite effects on growth, morphology, anchorage dependence, and tumorigenicity.
    J Biol Chem. 1993 Mar 25;268(9):6090-6 PMID: 8454583
  20. Protein kinase C isoenzymes: divergence in signal transduction?
    Biochem J. 1993 Apr 15;291 ( Pt 2):329-43 PMID: 8484714
  21. Cloning and expression of a protein kinase C-regulated chloride channel abundantly expressed in rat brain neuronal cells.
    Neuron. 1994 Mar;12(3):597-604 PMID: 8155321
  22. Activation of a Cl- current by hypotonic volume increase in human endothelial cells.
    J Gen Physiol. 1994 May;103(5):787-805 PMID: 7913485
  23. Correlation between protein kinase C binding proteins and substrates in REF52 cells.
    Cell Growth Differ. 1994 May;5(5):495-502 PMID: 8049156
  24. Protein kinase C-mediated phosphorylation of the human multidrug resistance P-glycoprotein regulates cell volume-activated chloride channels.
    EMBO J. 1995 Jan 3;14(1):68-75 PMID: 7828597
  25. Role of cell volume and protein kinase C in regulation of a Cl- conductance in single proximal tubule cells of Rana temporaria.
    J Physiol. 1994 Oct 1;480 ( Pt 1):1-7 PMID: 7853214
  26. PKC-sensitive Cl- channels associated with ciliary epithelial homologue of pICln.
    Am J Physiol. 1995 Mar;268(3 Pt 1):C572-9 PMID: 7534980
  27. Volume-activated, gadolinium-sensitive whole-cell currents in single proximal cells of frog kidney.
    Pflugers Arch. 1994 Nov;429(1):98-106 PMID: 7535920
  28. Protein kinase C and lipid signaling for sustained cellular responses.
    FASEB J. 1995 Apr;9(7):484-96 PMID: 7737456
  29. Activation of extracellular signal-regulated kinases Erk-1 and Erk-2 by cell swelling in H4IIE hepatoma cells.
    Biochem J. 1995 Jul 1;309 ( Pt 1):13-7 PMID: 7619047
  30. Membrane mechanisms and intracellular signalling in cell volume regulation.
    Int Rev Cytol. 1995;161:173-262 PMID: 7558691
  31. Microinjection of Rho and Rac into quiescent Swiss 3T3 cells.
    Methods Enzymol. 1995;256:313-20 PMID: 7476447
  32. Characterization of a volume-sensitive chloride current in rat osteoblast-like (ROS 17/2.8) cells.
    J Physiol. 1995 Jun 15;485 ( Pt 3):671-82 PMID: 7562609
  33. Activation of the plasma membrane chloride channel by protein kinase C in isolated guinea-pig hepatocytes.
    J Physiol. 1995 Sep 1;487 ( Pt 2):379-94 PMID: 8558471
  34. Volume-sensitive chloride channels associated with human cervical carcinogenesis.
    Cancer Res. 1995 Dec 15;55(24):6077-83 PMID: 8521396
  35. Volume-activated chloride currents in pancreatic duct cells.
    J Membr Biol. 1995 Sep;147(2):173-83 PMID: 8568853
  36. Activation of the volume-sensitive chloride current in vascular endothelial cells requires a permissive intracellular Ca2+ concentration.
    Pflugers Arch. 1996 Jan;431(3):467-9 PMID: 8584445
  37. Volume-sensitive chloride channels in the primary culture cells of human cervical carcinoma.
    Biochim Biophys Acta. 1996 Mar 1;1315(2):138-44 PMID: 8608171
  38. The role of cellular hydration in the regulation of cell function.
    Biochem J. 1996 Feb 1;313 ( Pt 3):697-710 PMID: 8611144
  39. Cellular and molecular physiology of volume-sensitive anion channels.
    Am J Physiol. 1996 Mar;270(3 Pt 1):C711-30 PMID: 8638650
  40. Identification of a major protein kinase C-binding protein and substrate in rat embryo fibroblasts. Decreased expression in transformed cells.
    J Biol Chem. 1996 Mar 15;271(11):6417-22 PMID: 8626441
  41. Modulation of the hyperpolarization-activated Cl- current in human intestinal T84 epithelial cells by phosphorylation.
    J Physiol. 1996 Jan 1;490 ( Pt 1):115-28 PMID: 8745282
  42. Chloride-channel block inhibits T lymphocyte activation and signalling.
    Cell Signal. 1996 Feb;8(2):141-9 PMID: 8730516
  43. Protein kinase C isozymes and substrates.
    Curr Opin Cell Biol. 1996 Apr;8(2):168-73 PMID: 8791416
  44. Involvement of protein kinase Cepsilon in the stimulation of anionic amino acid transport in cultured human fibroblasts.
    J Biol Chem. 1996 Oct 18;271(42):26124-30 PMID: 8824256
  45. Volume-activated Cl- channels.
    Gen Pharmacol. 1996 Oct;27(7):1131-40 PMID: 8981057
  46. Regulation of protein kinase C.
    Curr Opin Cell Biol. 1997 Apr;9(2):161-7 PMID: 9069266
  47. Volume expansion-sensing outward-rectifier Cl- channel: fresh start to the molecular identity and volume sensor.
    Am J Physiol. 1997 Sep;273(3 Pt 1):C755-89 PMID: 9316396
  48. Functional significance of cell volume regulatory mechanisms.
    Physiol Rev. 1998 Jan;78(1):247-306 PMID: 9457175
Article Info
Journal
The Journal of physiology
Abbr.
J Physiol
ISSN
0022-3751
DOI
10.1111/j.1469-7793.1998.435be.x
Email
chougyn@mail.ncku.edu.tw
Published
1998-10-15
Pages
435-48
Language
English
Region
England
NLM ID
0266262
PMCID
PMC2231219
Subset
IM
External Links
PubMed source DOI: 10.1111/j.1469-7793.1998.435be.x PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2