Abstract
The role of protein kinase C (PKC) in in vitro invasiveness of four different human renal cell carcinoma (RCC) cell lines of the clear cell type was investigated. Different PKC-inhibitors markedly inhibited invasiveness of the highly invasive cell lines, suggesting an invasion-promoting role of PKC in human RCC. Analysis of PKC-isoenzyme expression by protein fractionation and immunoblotting revealed that all cell lines expressed PKC-α, -ɛ, -ζ, -μ and -ι as known from normal kidney tissue. Interestingly, PKC-δ, known to be expressed by normal kidney epithelial cells of the rat, was absent on protein and RNA levels in all RCC cell lines investigated and in normal human kidney epithelial cells. PKC-ɛ expression levels correlated positively with a high proliferation activity, but no obvious correlation between expression levels of distinct PKC-isoenzymes and in vitro invasiveness was observed. However, by immunofluorescence microscopy, membrane localisation of PKC-α and PKC-ɛ reflecting activation of the enzymes, was associated with a highly invasive potential. In conclusion, our results suggest a role for PKC in invasion of human RCCs and might argue in favour of a particular role of PKC-α and PKC-ɛ. Our results further suggest that organ-specific expression patterns of PKC-isoenzymes are not necessarily conserved during evolution. © 2000 Cancer Research Campaign
Keywords: protein kinase C, renal cell carcinoma, invasion
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- Anglard P., Tory K., Brauch H., Weiss G. H., Latif F., Merino M. J., Lerman M. I., Zbar B., Linehan W. M. Molecular analysis of genetic changes in the origin and development of renal cell carcinoma. Cancer Res. 1991 Feb 15;51(4):1071–1077. [PubMed] [Google Scholar]
- Bareggi R., Grill V., Zweyer M., Narducci P., Martelli A. M. Distribution of the extended family of protein kinase C isoenzymes in fetal organs of mice: an immunohistochemical study. Cell Tissue Res. 1995 Jun;280(3):617–625. doi: 10.1007/BF00318364. [DOI] [PubMed] [Google Scholar]
- Blobe G. C., Obeid L. M., Hannun Y. A. Regulation of protein kinase C and role in cancer biology. Cancer Metastasis Rev. 1994 Dec;13(3-4):411–431. doi: 10.1007/BF00666107. [DOI] [PubMed] [Google Scholar]
- Borner C., Ueffing M., Jaken S., Parker P. J., Weinstein I. B. Two closely related isoforms of protein kinase C produce reciprocal effects on the growth of rat fibroblasts. Possible molecular mechanisms. J Biol Chem. 1995 Jan 6;270(1):78–86. doi: 10.1074/jbc.270.1.78. [DOI] [PubMed] [Google Scholar]
- Buchner K. Protein kinase C in the transduction of signals toward and within the cell nucleus. Eur J Biochem. 1995 Mar 1;228(2):211–221. [PubMed] [Google Scholar]
- Budworth J., Gescher A. Differential inhibition of cytosolic and membrane-derived protein kinase C activity by staurosporine and other kinase inhibitors. FEBS Lett. 1995 Apr 3;362(2):139–142. doi: 10.1016/0014-5793(95)00227-z. [DOI] [PubMed] [Google Scholar]
- Cacace A. M., Guadagno S. N., Krauss R. S., Fabbro D., Weinstein I. B. The epsilon isoform of protein kinase C is an oncogene when overexpressed in rat fibroblasts. Oncogene. 1993 Aug;8(8):2095–2104. [PubMed] [Google Scholar]
- Disatnik M. H., Winnier A. R., Mochly-Rosen D., Arteaga C. L. Distinct responses of protein kinase C isozymes to c-erbB-2 activation in SKBR-3 human breast carcinoma cells. Cell Growth Differ. 1994 Aug;5(8):873–880. [PubMed] [Google Scholar]
- Dong L., Stevens J. L., Fabbro D., Jaken S. Protein kinase C isozyme expression and down-modulation in growing, quiescent, and transformed renal proximal tubule epithelial cells. Cell Growth Differ. 1994 Aug;5(8):881–890. [PubMed] [Google Scholar]
- Dong L., Stevens J. L., Jaken S. Transformation-sensitive localization of alpha-protein kinase C at cell-cell contacts in rat renal proximal tubule epithelial cells. Cell Growth Differ. 1993 Oct;4(10):793–798. [PubMed] [Google Scholar]
- Druck T., Kastury K., Hadaczek P., Podolski J., Toloczko A., Sikorski A., Ohta M., LaForgia S., Lasota J., McCue P. Loss of heterozygosity at the familial RCC t(3;8) locus in most clear cell renal carcinomas. Cancer Res. 1995 Nov 15;55(22):5348–5353. [PubMed] [Google Scholar]
- Engers R., Gerharz C. D., Donner A., Mrzyk S., Krause-Paulus R., Petek O., Gabbert H. E. In vitro invasiveness of human epithelioid-sarcoma cell lines: association with cell motility and inverse correlation with the expression of tissue inhibitor of metalloproteinases. Int J Cancer. 1999 Jan 29;80(3):406–412. doi: 10.1002/(sici)1097-0215(19990129)80:3<406::aid-ijc12>3.0.co;2-l. [DOI] [PubMed] [Google Scholar]
- Engers R., Gerharz C. D., Moll R., Pohl A., Sarbia M., Gabbert H. E. Interclonal heterogeneity in a human epithelioid-sarcoma cell line (GRU-1). Int J Cancer. 1994 Nov 15;59(4):548–553. doi: 10.1002/ijc.2910590419. [DOI] [PubMed] [Google Scholar]
- Gabbert H. E., Gerharz C. D., Biesalski H. K., Engers R., Luley C. Terminal differentiation and growth inhibition of a rat rhabdomyosarcoma cell line (BA-HAN-1C) in vitro after exposure to retinoic acid. Cancer Res. 1988 Sep 15;48(18):5264–5269. [PubMed] [Google Scholar]
- Gerharz C. D., Moll R., Störkel S., Ramp U., Thoenes W., Gabbert H. E. Ultrastructural appearance and cytoskeletal architecture of the clear, chromophilic, and chromophobe types of human renal cell carcinoma in vitro. Am J Pathol. 1993 Mar;142(3):851–859. [PMC free article] [PubMed] [Google Scholar]
- Goldstein D. R., Cacace A. M., Weinstein I. B. Overexpression of protein kinase C beta 1 in the SW480 colon cancer cell line causes growth suppression. Carcinogenesis. 1995 May;16(5):1121–1126. doi: 10.1093/carcin/16.5.1121. [DOI] [PubMed] [Google Scholar]
- Goodnight J. A., Mischak H., Kolch W., Mushinski J. F. Immunocytochemical localization of eight protein kinase C isozymes overexpressed in NIH 3T3 fibroblasts. Isoform-specific association with microfilaments, Golgi, endoplasmic reticulum, and nuclear and cell membranes. J Biol Chem. 1995 Apr 28;270(17):9991–10001. doi: 10.1074/jbc.270.17.9991. [DOI] [PubMed] [Google Scholar]
- Hoelting T., Duh Q. Y., Clark O. H., Herfarth C. Tamoxifen antagonizes proliferation and invasion of estrogen receptor-negative metastatic follicular thyroid cancer cells via protein kinase C. Cancer Lett. 1996 Feb 27;100(1-2):89–93. doi: 10.1016/0304-3835(95)04074-9. [DOI] [PubMed] [Google Scholar]
- Huppi K., Siwarski D., Goodnight J., Mischak H. Assignment of the protein kinase C delta polypeptide gene (PRKCD) to human chromosome 3 and mouse chromosome 14. Genomics. 1994 Jan 1;19(1):161–162. doi: 10.1006/geno.1994.1028. [DOI] [PubMed] [Google Scholar]
- Jaken S. Protein kinase C isozymes and substrates. Curr Opin Cell Biol. 1996 Apr;8(2):168–173. doi: 10.1016/s0955-0674(96)80062-7. [DOI] [PubMed] [Google Scholar]
- Johannes F. J., Prestle J., Dieterich S., Oberhagemann P., Link G., Pfizenmaier K. Characterization of activators and inhibitors of protein kinase C mu. Eur J Biochem. 1995 Jan 15;227(1-2):303–307. doi: 10.1111/j.1432-1033.1995.tb20389.x. [DOI] [PubMed] [Google Scholar]
- Johannes F. J., Prestle J., Eis S., Oberhagemann P., Pfizenmaier K. PKCu is a novel, atypical member of the protein kinase C family. J Biol Chem. 1994 Feb 25;269(8):6140–6148. [PubMed] [Google Scholar]
- Kiley S. C., Parker P. J. Differential localization of protein kinase C isozymes in U937 cells: evidence for distinct isozyme functions during monocyte differentiation. J Cell Sci. 1995 Mar;108(Pt 3):1003–1016. doi: 10.1242/jcs.108.3.1003. [DOI] [PubMed] [Google Scholar]
- Kovacs G., Erlandsson R., Boldog F., Ingvarsson S., Müller-Brechlin R., Klein G., Sümegi J. Consistent chromosome 3p deletion and loss of heterozygosity in renal cell carcinoma. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1571–1575. doi: 10.1073/pnas.85.5.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liu J. P. Protein kinase C and its substrates. Mol Cell Endocrinol. 1996 Jan 15;116(1):1–29. doi: 10.1016/0303-7207(95)03706-3. [DOI] [PubMed] [Google Scholar]
- Mapelli E., Banfi P., Sala E., Sensi M., Supino R., Zunino F., Gambetta R. A. Effect of protein kinase C inhibitors on invasiveness of human melanoma clones expressing different levels of protein kinase C isoenzymes. Int J Cancer. 1994 Apr 15;57(2):281–286. doi: 10.1002/ijc.2910570225. [DOI] [PubMed] [Google Scholar]
- Mareel M. M. Recent aspects of tumor invasiveness. Int Rev Exp Pathol. 1980;22:65–129. [PubMed] [Google Scholar]
- Marte B. M., Meyer T., Stabel S., Standke G. J., Jaken S., Fabbro D., Hynes N. E. Protein kinase C and mammary cell differentiation: involvement of protein kinase C alpha in the induction of beta-casein expression. Cell Growth Differ. 1994 Mar;5(3):239–247. [PubMed] [Google Scholar]
- Meyer T., Regenass U., Fabbro D., Alteri E., Rösel J., Müller M., Caravatti G., Matter A. A derivative of staurosporine (CGP 41 251) shows selectivity for protein kinase C inhibition and in vitro anti-proliferative as well as in vivo anti-tumor activity. Int J Cancer. 1989 May 15;43(5):851–856. doi: 10.1002/ijc.2910430519. [DOI] [PubMed] [Google Scholar]
- Morita R., Ishikawa J., Tsutsumi M., Hikiji K., Tsukada Y., Kamidono S., Maeda S., Nakamura Y. Allelotype of renal cell carcinoma. Cancer Res. 1991 Feb 1;51(3):820–823. [PubMed] [Google Scholar]
- Mulders P., Figlin R., deKernion J. B., Wiltrout R., Linehan M., Parkinson D., deWolf W., Belldegrun A. Renal cell carcinoma: recent progress and future directions. Cancer Res. 1997 Nov 15;57(22):5189–5195. [PubMed] [Google Scholar]
- Prestle J., Pfizenmaier K., Brenner J., Johannes F. J. Protein kinase C mu is located at the Golgi compartment. J Cell Biol. 1996 Sep;134(6):1401–1410. doi: 10.1083/jcb.134.6.1401. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schwartz G. K., Jiang J., Kelsen D., Albino A. P. Protein kinase C: a novel target for inhibiting gastric cancer cell invasion. J Natl Cancer Inst. 1993 Mar 3;85(5):402–407. doi: 10.1093/jnci/85.5.402. [DOI] [PubMed] [Google Scholar]
- Selbie L. A., Schmitz-Peiffer C., Sheng Y., Biden T. J. Molecular cloning and characterization of PKC iota, an atypical isoform of protein kinase C derived from insulin-secreting cells. J Biol Chem. 1993 Nov 15;268(32):24296–24302. [PubMed] [Google Scholar]
- Toullec D., Pianetti P., Coste H., Bellevergue P., Grand-Perret T., Ajakane M., Baudet V., Boissin P., Boursier E., Loriolle F. The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C. J Biol Chem. 1991 Aug 25;266(24):15771–15781. [PubMed] [Google Scholar]
- Ways D. K., Kukoly C. A., deVente J., Hooker J. L., Bryant W. O., Posekany K. J., Fletcher D. J., Cook P. P., Parker P. J. MCF-7 breast cancer cells transfected with protein kinase C-alpha exhibit altered expression of other protein kinase C isoforms and display a more aggressive neoplastic phenotype. J Clin Invest. 1995 Apr;95(4):1906–1915. doi: 10.1172/JCI117872. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wetsel W. C., Khan W. A., Merchenthaler I., Rivera H., Halpern A. E., Phung H. M., Negro-Vilar A., Hannun Y. A. Tissue and cellular distribution of the extended family of protein kinase C isoenzymes. J Cell Biol. 1992 Apr;117(1):121–133. doi: 10.1083/jcb.117.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
