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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 2001 Aug;92(8):896–903. doi: 10.1111/j.1349-7006.2001.tb01178.x

Anticancer Drug‐mediated Induction of Multidrug Resistance‐associated Genes and Protein Kinase C Isozymes in the T‐Lymphoblastoid Cell Line CCRF‐CEM and in Blasts from Patients with Acute Lymphoblastic Leukemias

James F Beck 1,✉, Dorothee Brügger 2, Klaus Brischwein 1, Chao Liu 2, Peter Bader 2, Dietrich Niethammer 2, Volker Gekeler 3
PMCID: PMC5926830  PMID: 11509123

Abstract

The major determinants mediating drug resistance in acute lymphoblastic leukemias (ALL) unresponsive to chemotherapy, are still unclear. For example, it is still unknown whether selection or induction processes are responsible for drug resistance here or whether protein kinase C (PKC) isozymes contribute to the resistant phenotype. Therefore, inducibility of resistance factors or PKC isozymes genes was examined in CCRF‐CEM cells treated with diverse anticancer drugs‐ adriamycin, camptothecin, etoposide or vincristine‐at sublethal concentrations for 24 h. MDR1, MRP1, LRP and PKC isozyme α, β1, β2, ε, ι, η, θ, ζ gene expression was determined by cDNA‐PCR. We found significant dose‐dependent, mostly combined, induction of the MDR1, MRP1 and LRP genes. Significantly enhanced gene expression of the majority of PKC isozyme genes was found after treatment with camptothecin. PKCζ was upregulated throughout by each anticancer drug applied in this setting. A series of selected CCRF‐CEM‐derived multidrug resistance (MDR) sublines also showed enhanced expression of the PKC isozymes compared to the parental cell line. MDR1 and PKCη gene expression levels were correlated highly significantly. Blasts from two patients with ALL during the first week of monotherapy with steroids revealed combined induction of the MDR1, multidrug resistance‐associated protein 1 (MRP1), lung cancer resistance‐related protein (LRP) and most PKC isozymes, predominantly PKCζ. Another patient with T‐ALL, who failed to respond to four months of intensive chemotherapy, showed an enhanced MRP1 gene expression combined with markedly overexpression of PKCη and PKCθ. Furthermore, the camptothecin and etoposide‐mediated induction of resistance factors in the CCRF‐CEM cell line could be suppressed by staurosporine, a rather unspecific inhibitor of protein kinases. However, selective inhibitors of PKC isozymes (bisindolylmaleimide GÖ 6850, indolocarbazole GÖ 6976) produced no significant effects here. Therefore, the PKC isozymes η, θ and ζ are of interest as potential targets to overcome drug resistance in ALL.

Keywords: Acute lymphoblastic leukemia, Anticancer drugs, Gene induction, MDR, Protein kinase C

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References

  • 1.Germann , U. A.P‐Glycoprotein‐a mediator of multidrug resistance in tumour cells . Eur. J. Cancer. , 32A , 927 – 944 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 2.Loe , D. W. , Deeley , R. G. and Cole , S. P.Biology of the multidrug resistance‐associated protein, MRP . Eur. J. Cancer. , 32A , 945 – 957 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 3.Izquierdo , M. A. , Scheffer , G. L. , Flens , M. J. , Schroeijers , A. B. , van der Valk , P. and Scheper , R. J.Major vault protein LRP‐related multidrug resistance . Eur. J. Cancer. , 32A , 979 – 984 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 4.Gekeler , V. , Frese , G. , Noller , A. , Handgretinger , R. , Wilisch , A. , Schmidt , H. , Muller , C. P. , Dopfer , R. , Klingebiel , T. , Diddens , H. , Probst , H. and Niethammer , D . Mdrl/P‐glycoprotein, topoisomerase, and glutathione‐S‐transferase Jt gene expression in primary and relapsed state adult and childhood leukaemias . Br. J. Cancer , 66 , 507 – 517 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.Beck , J. , Niethammer , D. and Gekeler , V . High mdrl‐ and mrp‐, but low topoisomerase IIα‐gene expression in B‐cell chronic lymphocytic leukaemias . Cancer Lett. , 86 , 135 – 142 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 6.Beck , J. , Handgretinger , R. , Dopfer , R. , Klingebiel , T. , Niethammer , D. and Gekeler , V . Expression of MDR1, MRP, topoisomerase IIα/P, and cyclin A in primary or relapsed states of acute lymphoblastic leukaemias . Br. J. Haematol. , 89 , 356 – 363 ( 1995. ). [DOI] [PubMed] [Google Scholar]
  • 7.Burt , R. K. , Garfield , S. , Johnson , K. and Thorgeirsson , S. S . Transformation of rat liver epithelial cells with v‐H‐ras or v‐raf causes expression of MDR‐1, glutathione‐S‐trans‐ferase‐P and increased resistance to cytotoxic chemicals . Carcinogenesis , 9 , 2329 – 2332 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 8.Beck , J. , Niethammer , D. and Gekeler , V . MDR1, MRP, topoisomerase IIα/β, and cyclin A gene expression in acute and chronic leukemias . Leukemia , 10 ( Suppl. 3 ), 39 – 45 ( 1996. ). [PubMed] [Google Scholar]
  • 9.Beck , J. , Bohnet , B. , Brügger , D. , Bader , P. , Dietl , J. , Scheper , R. J. , Kandolf , R. , Liu , C. , Niethammer , D. and Gekeler , V . Multiple gene expression analysis reveals distinct differences between G2 and G3 breast cancers, and correlations of PKCη with MDR1, MRP and LRP gene expression . Br. J. Cancer , 77 , 87 – 91 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Fairchild , C. R. , Ivy , S. P. , Rushmore , T. , Lee , G. , Koo , P. , Goldsmith , M. E. , Myers , C. E. , Farber , E. and Cowan , K. H . Carcinogen‐induced mdr overexpression is associated with xenobiotic resistance in rat preneoplastic liver nodules and hepatocellular carcinomas . Proc. Natl. Acad. Sci. USA , 84 , 7701 – 7705 ( 1987. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Burt , R. K. and Thorgeirsson , S. S . Coinduction of MDR‐1 multidrug‐resistance and cytochrome P‐450 genes in rat liver by xenobiotics . J. Natl. Cancer Inst. , 80 , 1383 – 1386 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 12.Gekeler , V. , Frese , G. , Diddens , H. and Probst , H . Expression of a P‐glycoprotein gene is inducible in a multidrug‐resistant human leukemia cell line . Biochem. Biophys. Res. Commun. , 155 , 754 – 760 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 13.Kohno , K. , Sato , S. , Takano , H. , Matsuo , K. and Kuwano , M . The direct activation of human multidrug resistance gene (MDR1) by anticancer agents . Biochem. Biophys. Res. Commun. , 165 , 1415 – 1421 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 14.Chaudhary , P. M and Roninson , I. B . Induction of multi‐drug resistance in human cells by transient exposure to different chemotherapeutic drugs . J. Natl. Cancer Inst. , 85 , 632 – 639 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 15.Licht , T. , Fiebig , H. H. , Bross , K. J. , Herrmann , F. , Berger , D. P. , Shoemaker , R. and Mertelsmann , R.Induction of multiple‐drug resistance during anti‐neoplastic chemotherapy in vitro . Int. J. Cancer , 49 , 630 – 637 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 16.Gekeler , V. , Beck , J. , Noller , A. , Wilisch , A. , Frese , G. , Neumann , M. , Handgretinger , R. , Ehninger , G. , Probst , H. and Niethammer , D . Drug‐induced changes in the expression of MDR‐associated genes: investigations on cultured cell lines and chemotherapeutically treated leukemias . Ann. Hematol , 69 , 19 – 24 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 17.Fine , R. L. , Patel , J. and Chabner , B. A . Phorbol esters induce multidrug resistance in human breast cancer cells . Proc. Natl. Acad. Sci. USA , 85 , 582 – 586 ( 1988. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Mickley , L. A. , Bates , S. E. , Richert , N. D. , Currier , S. , Tanaka , S. , Foss , F. , Rosen , N. and Fojo , A. T . Modulation of the expression of a multidrug resistance gene (mdr‐l/P‐glycoprotein) by differentiating agents . J. Biol. Chem. , 264 , 18031 – 18040 ( 1989. ). [PubMed] [Google Scholar]
  • 19.Chaudhary , P. M. and Roninson , I. B . Activation of MDR1 (P‐glycoprotein) gene expression in human cells by protein kinase C agonists . Oncol. Res. , 4 , 281 – 290 ( 1992. ). [PubMed] [Google Scholar]
  • 20.Chin , K. V. , Tanaka , S. , Darlington , G. , Pastan , I. and Gottesman , M. M . Heat shock and arsenite increase expression of the multidrug resistance (MDR1) gene in human renal carcinoma cells . J. Biol. Chem. , 265 , 221 – 226 ( 1990. ). [PubMed] [Google Scholar]
  • 21.Miyazaki , M. , Kohno , K. , Uchiumi , T. , Tanimura , H. , Matsuo , K. , Nasu , M. and Kuwano , M . Activation of human multidrug resistance‐1 gene promoter in response to heat shock stress . Biochem. Biophys. Res. Commun. , 187 , 677 – 684 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 22.Bates , S. E. , Lee , J. S. , Dickstein , B. , Spolyar , M. and Fojo , A. T . Differential modulation of P‐glycoprotein transport by protein kinase inhibition . Biochemistry , 32 , 9156 – 9164 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 23.Sampson , K. E. , Wolf , C. L. and Abraham , I . Staurosporine reduces P‐glycoprotein expression and modulates multidrug resistance . Cancer Lett. , 68 , 7 – 14 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 24.Uchiumi , T. , Kohno , K. , Tanimura , H. , Hidaka , K. , Asakuno , K. , Abe , H. , Uchida , Y. and Kuwano , M . Involvementof proteinkinase in environmental stress‐induced activation of human multidrug resistance 1 (MDR1) gene promoter . FEBS Lett. , 326 , 11 – 16 ( 1993. ). [DOI] [PubMed] [Google Scholar]
  • 25.Toullec , D. , Pianetti , P. , Coste , H. , Bellevergue , P. , Grand , P. T. , Ajakane , M. , Baudet , V. , Boissin , P. , Boursier , E. , Loriolle , F. , Duhamel , L. , Charon , D. and Kirilovsky , J . The bisindolylmaleimide GF 109203X is a potent and selective inhibitor of protein kinase C . J. Biol. Chem. , 266 , 15771 – 15781 ( 1991. ). [PubMed] [Google Scholar]
  • 26.Martiny‐Baron , G. , Kazanietz , M. G. , Mischak , H. , Blumberg , P. M. , Kochs , G. , Hug , H. , Marme , D. and Schächtele , C . Selective inhibition of PKC isozymes by the indolocarbazole Gö 6976 . J. Biol. Chem. , 268 , 9194 – 9197 ( 1993. ). [PubMed] [Google Scholar]
  • 27.Grunicke , H. , Hofmann , J. , Utz , I. and Überall , F . Role of protein kinases in antitumor drug resistance . Ann. Hema-tol , 69 , 1 – 6 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 28.Kimmig , A. , Gekeler , V. , Neumann , M. , Frese , G. , Hand‐gretinger , R. , Kardos , G. , Diddens , H. and Niethammer , D . Susceptibility of multidrug‐resistant human leukemia cell lines to human interleukin 2‐activated killer cells . Cancer Res. , 50 , 6793 – 6799 ( 1990. ). [PubMed] [Google Scholar]
  • 29.Riehm , H . Multicenter trial of the German Society for Pediatric Oncology and Hematology ALL‐BFM‐95 for treatment of children with acute lymphoblastic leukemia . Treatment Protocol. , 1 , 1 – 101 ( 1995. ). [Google Scholar]
  • 30.Tamaoki , T. , Nomoto , H. , Takahashi , I. , Kato , Y. , Morimoto , M. and Tomita , F.Staurosporine, a potent inhibitor of phospholipid/Ca++ ‐dependent protein kinase . Biochem. Biophys. Res. Commun. , 135 , 397 – 402 ( 1986. ). [DOI] [PubMed] [Google Scholar]
  • 31.Sato , W. , Yusa , K. , Naito , M. and Tsuruo , T . Staurosporine, a potent inhibitor of C‐kinase, enhances drug accumulation in multidrug‐resistant cells . Biochem. Biophys. Res. Commun. , 173 , 1252 – 1257 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 32.Chirgwin , J. M. , Przybyla , A. E. , MacDonald , R. J. and Rutter , W. J . Isolation of biological active ribonucleic acid from sources enriched in ribonuclease . Biochemistry , 18 , 5294 – 5299 ( 1979. ). [DOI] [PubMed] [Google Scholar]
  • 33.Gekeler , V. , Boer , R. , Überall , F. , Ise , W. , Schubert , C. , Utz , I. , Hofmann , J. , Sanders , K. H. , Schächtele , C. , Klemm , K. and Grunicke , H . Effects of the selective bisindolylmaleimide protein kinase C inhibitor GF109203X on P‐glycoprotein mediated multidrug resistance . Br. J. Cancer , 74 , 897 – 905 ( 1996. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Mischak , H. , Kolchm , W. , Goodnight , J. , Davidson , W. F. , Rapp , U. , Rose‐John , S. and Mushinski , J. F . Expression of protein kinase C genes in hematopoietic cells is cell‐type and B cell‐differentiation stage specific . J. Immunol. , 147 , 3981 – 3987 ( 1991. ). [PubMed] [Google Scholar]
  • 35.Chen , A. Y. , Yu , C. , Potmesil , M. , Wall , M. E. , Wani , M. C. and Liu , L. F . Camptothecin overcomes MDRl‐medi‐ated resistance in human KB carcinoma cells . Cancer Res. , 51 , 6039 – 6044 ( 1991. ). [PubMed] [Google Scholar]
  • 36.Ma , J. , Maliepaard , M. , Nooter , K. , Loos , W. J. , Kolker , H. J. , Verweij , J. , Stoter , G. and Schellens , J. H . Reduced cellular accumulationof topotecan: a novel mechanism of resistance in a human ovarian cancer cell line . Br. J. Cancer , 77 , 1645 – 1652 ( 1998. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Komarov , P. G. , Shtil , A. A. , Holian , O. , Tee , L. , Buckingham , L. , Mechetner , E. B. , Roninson , I. B. and Coon , J. S . Activation of the LRP (lung resistance‐related protein) gene by short‐term exposure of human leukemia cells to phorbol ester and cytarabine . Oncol. Res. , 10 , 185 – 192 ( 1998. ). [PubMed] [Google Scholar]
  • 38.Beck , J. , Handgretinger , R. , Klingebiel , T. , Dopfer , R. , Schaich , M. , Ehninger , G. , Niethammer , D. and Gekeler , V . Expression of PKC isozyme and MDR‐associated genes in primary and relapsed state AML . Leukemia , 10 , 426 – 433 ( 1996. ). [PubMed] [Google Scholar]

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