Skip to main content
NCBI home page
Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1989 Apr;80(4):373–379. doi: 10.1111/j.1349-7006.1989.tb02322.x

Altered Expression of Epidermal Growth Factor Receptor Gene in a Classical Multidrug‐resistant Variant of a Human Cancer Cell Line, KB

Hiroshi Takano 1, Kimitoshi Kohno 1,✉, Norio Shiraishi 1, Shin‐ichi Sato 1, Kuni‐ichi Asoh 1, Miki Yakushinji 1, Mayumi Ono 1, Michihiko Kuwano 1
PMCID: PMC5917741  PMID: 2501254

Abstract

A variant clone resistant to high doses of colchicine (KB‐C1) derived from human cancer KB cell line is resistant to various anticancer agents. The KB‐C1 cells were much more resistant to epidermal growth factor and a chimeric toxin, EGF‐Pseudomottas exotoxin (PE), than the parental KB cells. KB‐C1 cells have decreased numbers of EGF‐receptors, though the affinity of the receptors is similar to that in the parental KB cells. A drug‐sensitive revertant (C1‐R2) partially recovered its EGF‐receptor activity. Northern blot analysis showed a decreased level of EGF‐receptor mRNA in KB‐C1 cells, while the multidrug‐resistance gene, mdr‐1, was expressed at very high levels in KB‐C1 cells, but not in KB or C1‐R2 cells. The drug‐resistant cells were less tumorigenic than the parental cells when injected into nude mice. A decreased expression of EGF‐receptor in these cells may be one of the pleiotropic properties of multidrug‐resistant cells and may perhaps represent the basis for their reduced tumorigenicity.

Keywords: Multidrug resistance, Growth factor, Tumorigenicity

Full Text

The Full Text of this article is available as a PDF (810.4 KB).

REFERENCES

  • 1. ) Lane , M.Clinical problems of resistance to cancer chemotherapeutic agents . Fed. Proc. , 38 , 103 – 107 ( 1979. ). [PubMed] [Google Scholar]
  • 2. ) Ling , V. and Baker , R. M.Dominance of colchicine resistance in hybrid CHO cells . Somatic Cell Genet. , 4 , 193 – 200 ( 1978. ). [DOI] [PubMed] [Google Scholar]
  • 3. ) Debenham , P. G. , Kartner , N. , Siminovitch , L. , Riordan , J. R. and Ling , V.DNA mediated transfer of multidrug‐resistance and plasma membrane gloycoprotein expression . Mol. Cell Biol. , 2 , 881 – 889 ( 1982. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. ) Juliano , R. L. and Ling , V.A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants . Biochim. Biophys. Acta. 455 , 152 – 162 ( 1976. ). [DOI] [PubMed] [Google Scholar]
  • 5. ) Kartner , N. , Riordan , J. and Ling , V.Cell surface P‐glycoprotein associated with multidrug resistance in mammalian cell lines . Sience , 221 , 1285 – 1288 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 6. ) Riordan , J. R. and Ling , V.Purification of P‐glycoprotein from plasma membrane vesicles of Chinese hamster ovary cell mutants with reduced colchicine permeability . J. Biol. Chem. , 254 , 12701 – 12705 ( 1979. ). [PubMed] [Google Scholar]
  • 7. ) Gros , P. , Croop , J. and Housman , D.Mammalian multidrug‐resistance gene: complete cDNA sequence indicates strong homology to bacterial transport proteins . Cell , 47 , 371 – 380 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 8. ) Chen , C. J. , Chin , E. J. , Ueda , K. , Clark , P. D. , Pastan , I. , Gottesman , M. M. and Roninson , B. I.Internal duplication and homology with bacterial transport proteins in the mdr‐1 (P‐glycoprotein) gene from multidrug‐resistant human cells . Cell , 47 , 381 – 389 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 9. ) Shen , D. W. , Fojo , A. , Chin , J. E. , Roninson , I. B. , Richert , N. , Pastan , I. and Gottesman , M. M.Human multidrug‐resistant cell lines: increased mdr‐1 expression can precede gene amplification . Science , 232 , 643 – 645 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 10. ) Danks , M. K. , Metzger , D. W. , Ashmun , R. A. and Beck , W. T.Monoclonal antibodies to glycoproteins of Vinca alkaloid‐resistant human leukemic cells . Cancer Res. , 45 , 3220 – 3224 ( 1985. ). [PubMed] [Google Scholar]
  • 11. ) Beck , W. T. , Cirtain , C. M. , Ashmun , A. R. and Mirro , J.Differentiation and the multiple drug resistance phenotype in human leukemic cells . Cancer Res. , 46 , 4571 – 4575 ( 1986. ). [PubMed] [Google Scholar]
  • 12. ) Richert , N. , Akiyama , S. , Shen , D. , Gottesman , M. M. and Pastan , I.Multiply drug‐resistant human KB carcinoma cells have decreased amounts of a 75‐kDa glycoprotein . Proc. Natl. Acad. Sci. USA , 82 , 2330 – 2333 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13. ) Peterson , R. H. R , Beutler , W. J. and Biedler , J. L.Ganglioside composition of malignant and actinomycin D‐resistant nonmalignant Chinese hamster cells . Biochem. Pharmacol. , 28 , 579 – 582 ( 1979. ). [DOI] [PubMed] [Google Scholar]
  • 14. ) Biedler , L. J. , Riehm , H. , Peterson , H. E. R. and Spengler , A. B.Membrane‐mediated drug resistance and phenotypic reversion to normal growth behavior of Chinese hamster cells . J. Natl. Cancer Inst. , 55 , 671 – 677 ( 1975. ). [DOI] [PubMed] [Google Scholar]
  • 15. ) Meyers , B. M. , Merlnzzi , J. V. , Spengler , A. B. and Biedler , L. J.Epidermal growth factor receptor is increased in multidrug resistant Chinese hamster and mouse tumor cells . Proc. Natl. Acad. Sci. USA , 83 , 5521 – 5525 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16. ) Akiyama , S. , Fojo , A. , Hanover , J. A. , Pastan , I. and Gottesman , M. M.Isolation and genetic characterization of human KB cell line resistant to multiple drugs . Somatic Cell Mol. Genet. , 11 , 117 – 126 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 17. ) Shiraishi , N. , Akiyama , S. , Nakagawa , M. , Kobayashi , M. and Kuwano , M.Effect of bisbenzylisoquinoline (biscoclaurine) alkaloids on multidrug resistance in KB human cancer cells . Cancer Res. , 47 , 2413 – 2416 ( 1987. ). [PubMed] [Google Scholar]
  • 18. ) Shiraishi , N. , Akiyama , S. , Hagino , Y. , Kohno , K. , Kobayashi , M. , Kuwano , M. and Akiyama , S.Potentiation by a biscoclaurine alkaloid, cepharanthine, of the toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin in HeLa cells . Cancer Res. , 48 , 1307 – 1311 ( 1988. ). [PubMed] [Google Scholar]
  • 19. ) Kuratomi , Y. , Akiyama , S. , Ono , M. , Shiraishi , N. , Shimada , T. , Ohkuma , S. and Kuwano , M.Thioridazine enhances lysosomal accumulation of epidermal growth factor and toxicity of conjugates of epidermal growth factor with Pseudomonas exotoxin . Exp. Cell Res. , 162 , 436 – 447 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 20. ) Das , M. , Miyakawa , T. , Fox , C. F. , Pruso , R. M. , Aharonov , A. and Herschman , H. R.Specific radiolabeling of a cell surface receptor for epidermal growth factor . Proc. Natl. Acad. Sci. USA , 74 , 2790 – 2794 ( 1977. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. ) Maniatis , T. , Fritsch , F. E. and Sambrook , J. “ Molecular Cloning: A Laboratory Manual ,” pp. 191 – 193 ( 1982. ). Cold Spring Harbor Laboratory; , Cold Spring Harbor , New York . [Google Scholar]
  • 22. ) Merlino , G. T. , Ishii , S. , Whang‐Peng , J. , Knutsen , T. , Xu , Y. H. , Clark , A. J. L. , Stratton , R. H. , Wilson , R. K. , Ma , D. P. , Roe , B. A. , Hunts , J. H. , Shimizu , N. and Pastan , I.Structure and localization of genes encoding aberrant and normal epidermal growth factor RNAs from A431 human carcinoma cells . Mol. Cell. Biol. , 5 , 1722 – 1734 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. ) Roninson , I. B. , Chin , J. E. , Choi , K. , Gros , P. , Housman , D. E. , Fojo , A. , Shen , D. W. , Gottesman , M. M. and Pastan , I.Isolation of the human mdr DNA sequences amplified in multidrug‐resistant KB carcinoma cells . Proc. Natl. Acad. Sci. USA , 83 , 4538 – 542 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24. ) Iijirma , N. S. , Hamada , H. , Reddy , P , and Kakunaga , T.Molecular structure of the human cytoplasmic β‐actin gene: interspecies homology of sequences in the introns . Proc. Natl. Acad. Sci. USA , 82 , 6133 – 6137 ( 1985. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. ) Feinberg , A. and Vogelstein , B.A technique for redio‐labeling DNA restriction endonuclease fragments to high specific activity . Anal. Biochem. , 132 , 6 – 13 ( 1983. ). [DOI] [PubMed] [Google Scholar]
  • 26. ) Shen , D. W. , Cardarelli , C. , Hwang , J. , Cornwell Richert , N. , Ishii , S. , Pastan , I. and Gottesman , M. M.Multiple drug‐resistant human KB carcinoma cells independently selected for high‐level resistance to colchicine, adriamycin, or vinblastine show changes in expression of specific proteins . J. Biol. Chem. , 261 , 7762 – 7770 ( 1986. ). [PubMed] [Google Scholar]
  • 27. ) Hwang , J. , Richert , N. , Pastan , I. and Gottesman , M. M.Mutant KB cells with decreased EGF receptor expression: biochemical characterization . J. Cell. Physiol. , 133 , 127 – 134 ( 1987. ). [DOI] [PubMed] [Google Scholar]
  • 28. ) Lyall , R. M. , Pastan , I. and Willingham , M. C.EGF induces receptor down‐regulation with no receptor recycling in KB cells . J. Cell. Physiol. , 122 , 166 – 170 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 29. ) Biedler , J. L. , Meyers , M. B. and Spengler , B. A.Cellular concomitants of multidrug resistance . In “ Mechanisms of Drug Resistance in Neoplastic Cells ,” ed , Woolley P. V., III and Tew K. D. , pp. 41 – 68 ( 1988. ). Academic Press, Inc. , New York . [Google Scholar]
  • 30. ) Ganapathi , R. , Grabowski , D. , Schmidt , H. , Bell , D. and Melia , M.Characterization in vitro and in vivo of progressively adriamycin‐resistant B16‐BL6 mouse melanoma cells . Cancer Res. , 47 , 3464 – 3468 ( 1987. ). [PubMed] [Google Scholar]
  • 31. ) Hirai , M. , Gamou , S. , Minoshima , S. and Shimizu , N.Two independent mechanisms for escaping epidermal growth factor mediated growth inhibition in epidermal growth factor receptor‐hyperproducing human tumors. J . Cell. Biol. , 107 , 791 – 799 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32. ) Shimizu , M. , Behzadian , A. M. and Shimizu , Y.Genetics of cell surface receptors for bioactive polypeptides: binding of epidermal growth factor is associated with the presence of human chromosome 7 in human‐mouse cell hybrids . Proc. Natl Acad. Sci. USA , 77 , 3600 – 3604 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. ) Koprowski , H. , Herlyn , M. , Balaban , G. , Parmiter , A. , Ross , A. and Nowell , P.Expression of the receptor for epidermal growth factor correlates with increased dosage of chromosome 7 in malignant melanoma . Somatic Cell Mol. Genet. , 11 , 297 – 301 ( 1985. ). [DOI] [PubMed] [Google Scholar]
  • 34. ) Slovak , L. M. , Hoeltge , A. G. and Trent , M. J.Cytogenetic alterations associated with the acquisition of doxorubicin resistance: possible significance of chromosome 7 alterations . Cancer Res. , 47 , 6646 – 6652 ( 1987. ). [PubMed] [Google Scholar]
  • 35. ) Collard , G. J. , van de Poll , M. , Schetter , A. , Roos , E. , Hopman , H. M. A. , Geurts van Kessel , H. M. A. and van Dongen , J. M. J.Location of genes involved in invasion and metastasis on human chromosome 7 . Cancer Res. , 47 , 6666 – 6670 ( 1987. ). [PubMed] [Google Scholar]

Articles from Japanese Journal of Cancer Research : Gann are provided here courtesy of Wiley

RESOURCES