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. 1996 Feb;40(2):419–425. doi: 10.1128/aac.40.2.419

Correlation between rhodamine 123 accumulation and azole sensitivity in Candida species: possible role for drug efflux in drug resistance.

F S Clark 1, T Parkinson 1, C A Hitchcock 1, N A Gow 1
PMCID: PMC163126  PMID: 8834890

Abstract

A wide variety of prokaryotic and eukaryotic cells exhibit a multidrug resistance (MDR) phenotype, indicating that resistance to potentially toxic compounds is mediated by their active efflux from the cell. We have sought to determine whether resistance to azoles in some strains of Candida species may be due in part to active drug efflux. Rhodamine 123 (Rh123) is a fluorescent compound that is transported by a wide variety of MDR cell types. We have shown that certain azole-resistant strains of Candida albicans, C. glabrata, and C. krusei accumulate less Rh123 than azole-susceptible ones. In C. albicans, Rh123 accumulation was growth phase and temperature dependent and was increased by proton uncouplers and by reserpine, an MDR modulator. This is consistent with an energy-dependent efflux mechanism for Rh123, mediated by an MDR transporter. In C. glabrata, but not in C. albicans, there was competition between Rh123 and fluconazole for efflux. Thus, in C. glabrata, Rh123 and fluconazole appear to be transported via a common MDR-like transporter, whereas in C. albicans, the Rh123 transporter does not appear to transport azoles.

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Selected References

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  1. Ambudkar S. V., Lelong I. H., Zhang J., Cardarelli C. O., Gottesman M. M., Pastan I. Partial purification and reconstitution of the human multidrug-resistance pump: characterization of the drug-stimulatable ATP hydrolysis. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8472–8476. doi: 10.1073/pnas.89.18.8472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Balzi E., Goffeau A. Genetics and biochemistry of yeast multidrug resistance. Biochim Biophys Acta. 1994 Aug 30;1187(2):152–162. doi: 10.1016/0005-2728(94)90102-3. [DOI] [PubMed] [Google Scholar]
  3. Balzi E., Goffeau A. Multiple or pleiotropic drug resistance in yeast. Biochim Biophys Acta. 1991 Mar 4;1073(2):241–252. doi: 10.1016/0304-4165(91)90128-4. [DOI] [PubMed] [Google Scholar]
  4. Balzi E., Wang M., Leterme S., Van Dyck L., Goffeau A. PDR5, a novel yeast multidrug resistance conferring transporter controlled by the transcription regulator PDR1. J Biol Chem. 1994 Jan 21;269(3):2206–2214. [PubMed] [Google Scholar]
  5. Ben-Yaacov R., Knoller S., Caldwell G. A., Becker J. M., Koltin Y. Candida albicans gene encoding resistance to benomyl and methotrexate is a multidrug resistance gene. Antimicrob Agents Chemother. 1994 Apr;38(4):648–652. doi: 10.1128/aac.38.4.648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bernal S. D., Lampidis T. J., Summerhayes I. C., Chen L. B. Rhodamine-123 selectively reduces clonal growth of carcinoma cells in vitro. Science. 1982 Dec 10;218(4577):1117–1119. doi: 10.1126/science.7146897. [DOI] [PubMed] [Google Scholar]
  7. Broxterman H. J., Pinedo H. M., Kuiper C. M., Kaptein L. C., Schuurhuis G. J., Lankelma J. Induction by verapamil of a rapid increase in ATP consumption in multidrug-resistant tumor cells. FASEB J. 1988 Apr;2(7):2278–2282. doi: 10.1096/fasebj.2.7.3350243. [DOI] [PubMed] [Google Scholar]
  8. Chaudhary P. M., Roninson I. B. Expression and activity of P-glycoprotein, a multidrug efflux pump, in human hematopoietic stem cells. Cell. 1991 Jul 12;66(1):85–94. doi: 10.1016/0092-8674(91)90141-k. [DOI] [PubMed] [Google Scholar]
  9. Cornwell M. M., Tsuruo T., Gottesman M. M., Pastan I. ATP-binding properties of P glycoprotein from multidrug-resistant KB cells. FASEB J. 1987 Jul;1(1):51–54. doi: 10.1096/fasebj.1.1.2886389. [DOI] [PubMed] [Google Scholar]
  10. Dexter D., Moye-Rowley W. S., Wu A. L., Golin J. Mutations in the yeast PDR3, PDR4, PDR7 and PDR9 pleiotropic (multiple) drug resistance loci affect the transcript level of an ATP binding cassette transporter encoding gene, PDR5. Genetics. 1994 Feb;136(2):505–515. doi: 10.1093/genetics/136.2.505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Efferth T., Löhrke H., Volm M. Reciprocal correlation between expression of P-glycoprotein and accumulation of rhodamine 123 in human tumors. Anticancer Res. 1989 Nov-Dec;9(6):1633–1637. [PubMed] [Google Scholar]
  12. Fath M. J., Kolter R. ABC transporters: bacterial exporters. Microbiol Rev. 1993 Dec;57(4):995–1017. doi: 10.1128/mr.57.4.995-1017.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ford J. M., Hait W. N. Pharmacology of drugs that alter multidrug resistance in cancer. Pharmacol Rev. 1990 Sep;42(3):155–199. [PubMed] [Google Scholar]
  14. Gallagher P. J., Bennett D. E., Henman M. C., Russell R. J., Flint S. R., Shanley D. B., Coleman D. C. Reduced azole susceptibility of oral isolates of Candida albicans from HIV-positive patients and a derivative exhibiting colony morphology variation. J Gen Microbiol. 1992 Sep;138(9):1901–1911. doi: 10.1099/00221287-138-9-1901. [DOI] [PubMed] [Google Scholar]
  15. Hamada H., Tsuruo T. Purification of the 170- to 180-kilodalton membrane glycoprotein associated with multidrug resistance. 170- to 180-kilodalton membrane glycoprotein is an ATPase. J Biol Chem. 1988 Jan 25;263(3):1454–1458. [PubMed] [Google Scholar]
  16. Hill B. T., Hosking L. K. Differential effectiveness of a range of novel drug-resistance modulators, relative to verapamil, in influencing vinblastine or teniposide cytotoxicity in human lymphoblastoid CCRF-CEM sublines expressing classic or atypical multidrug resistance. Cancer Chemother Pharmacol. 1994;33(4):317–324. doi: 10.1007/BF00685907. [DOI] [PubMed] [Google Scholar]
  17. Hitchcock C. A., Barrett-Bee K. J., Russell N. J. The lipid composition of azole-sensitive and azole-resistant strains of Candida albicans. J Gen Microbiol. 1986 Sep;132(9):2421–2431. doi: 10.1099/00221287-132-9-2421. [DOI] [PubMed] [Google Scholar]
  18. Hitchcock C. A., Pye G. W., Troke P. F., Johnson E. M., Warnock D. W. Fluconazole resistance in Candida glabrata. Antimicrob Agents Chemother. 1993 Sep;37(9):1962–1965. doi: 10.1128/aac.37.9.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Hitchcock C. A. Resistance of Candida albicans to azole antifungal agents. Biochem Soc Trans. 1993 Nov;21(4):1039–1047. doi: 10.1042/bst0211039. [DOI] [PubMed] [Google Scholar]
  20. Horio M., Gottesman M. M., Pastan I. ATP-dependent transport of vinblastine in vesicles from human multidrug-resistant cells. Proc Natl Acad Sci U S A. 1988 May;85(10):3580–3584. doi: 10.1073/pnas.85.10.3580. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Horsburgh C. R., Jr, Kirkpatrick C. H. Long-term therapy of chronic mucocutaneous candidiasis with ketoconazole: experience with twenty-one patients. Am J Med. 1983 Jan 24;74(1B):23–29. doi: 10.1016/0002-9343(83)90511-9. [DOI] [PubMed] [Google Scholar]
  22. Howell S. A., Mallet A. I., Noble W. C. A comparison of the sterol content of multiple isolates of the Candida albicans Darlington strain with other clinically azole-sensitive and -resistant strains. J Appl Bacteriol. 1990 Nov;69(5):692–696. doi: 10.1111/j.1365-2672.1990.tb01564.x. [DOI] [PubMed] [Google Scholar]
  23. Jancis E. M., Carbone R., Loechner K. J., Dannies P. S. Estradiol induction of rhodamine 123 efflux and the multidrug resistance pump in rat pituitary tumor cells. Mol Pharmacol. 1993 Jan;43(1):51–56. [PubMed] [Google Scholar]
  24. Johnson L. V., Walsh M. L., Chen L. B. Localization of mitochondria in living cells with rhodamine 123. Proc Natl Acad Sci U S A. 1980 Feb;77(2):990–994. doi: 10.1073/pnas.77.2.990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Juliano R. L., Ling V. A surface glycoprotein modulating drug permeability in Chinese hamster ovary cell mutants. Biochim Biophys Acta. 1976 Nov 11;455(1):152–162. doi: 10.1016/0005-2736(76)90160-7. [DOI] [PubMed] [Google Scholar]
  26. Juranka P. F., Zastawny R. L., Ling V. P-glycoprotein: multidrug-resistance and a superfamily of membrane-associated transport proteins. FASEB J. 1989 Dec;3(14):2583–2592. doi: 10.1096/fasebj.3.14.2574119. [DOI] [PubMed] [Google Scholar]
  27. Lomovskaya O., Lewis K. Emr, an Escherichia coli locus for multidrug resistance. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8938–8942. doi: 10.1073/pnas.89.19.8938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Miyauchi S., Komatsubara M., Kamo N. In archaebacteria, there is a doxorubicin efflux pump similar to mammalian P-glycoprotein. Biochim Biophys Acta. 1992 Oct 5;1110(2):144–150. doi: 10.1016/0005-2736(92)90351-l. [DOI] [PubMed] [Google Scholar]
  29. Neyfakh A. A., Bidnenko V. E., Chen L. B. Efflux-mediated multidrug resistance in Bacillus subtilis: similarities and dissimilarities with the mammalian system. Proc Natl Acad Sci U S A. 1991 Jun 1;88(11):4781–4785. doi: 10.1073/pnas.88.11.4781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Neyfakh A. A., Borsch C. M., Kaatz G. W. Fluoroquinolone resistance protein NorA of Staphylococcus aureus is a multidrug efflux transporter. Antimicrob Agents Chemother. 1993 Jan;37(1):128–129. doi: 10.1128/aac.37.1.128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Neyfakh A. A., Serpinskaya A. S., Chervonsky A. V., Apasov S. G., Kazarov A. R. Multidrug-resistance phenotype of a subpopulation of T-lymphocytes without drug selection. Exp Cell Res. 1989 Dec;185(2):496–505. doi: 10.1016/0014-4827(89)90318-2. [DOI] [PubMed] [Google Scholar]
  32. Odds F. C. Resistance of yeasts to azole-derivative antifungals. J Antimicrob Chemother. 1993 Apr;31(4):463–471. doi: 10.1093/jac/31.4.463. [DOI] [PubMed] [Google Scholar]
  33. Ouellette M., Légaré D., Papadopoulou B. Microbial multidrug-resistance ABC transporters. Trends Microbiol. 1994 Oct;2(10):407–411. doi: 10.1016/0966-842x(94)90620-3. [DOI] [PubMed] [Google Scholar]
  34. Parkinson T., Falconer D. J., Hitchcock C. A. Fluconazole resistance due to energy-dependent drug efflux in Candida glabrata. Antimicrob Agents Chemother. 1995 Aug;39(8):1696–1699. doi: 10.1128/aac.39.8.1696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pearce H. L., Safa A. R., Bach N. J., Winter M. A., Cirtain M. C., Beck W. T. Essential features of the P-glycoprotein pharmacophore as defined by a series of reserpine analogs that modulate multidrug resistance. Proc Natl Acad Sci U S A. 1989 Jul;86(13):5128–5132. doi: 10.1073/pnas.86.13.5128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pfaller M. A., Rinaldi M. G., Galgiani J. N., Bartlett M. S., Body B. A., Espinel-Ingroff A., Fromtling R. A., Hall G. S., Hughes C. E., Odds F. C. Collaborative investigation of variables in susceptibility testing of yeasts. Antimicrob Agents Chemother. 1990 Sep;34(9):1648–1654. doi: 10.1128/aac.34.9.1648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Prasad R., De Wergifosse P., Goffeau A., Balzi E. Molecular cloning and characterization of a novel gene of Candida albicans, CDR1, conferring multiple resistance to drugs and antifungals. Curr Genet. 1995 Mar;27(4):320–329. doi: 10.1007/BF00352101. [DOI] [PubMed] [Google Scholar]
  38. Rex J. H., Rinaldi M. G., Pfaller M. A. Resistance of Candida species to fluconazole. Antimicrob Agents Chemother. 1995 Jan;39(1):1–8. doi: 10.1128/aac.39.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ryley J. F., Wilson R. G., Barrett-Bee K. J. Azole resistance in Candida albicans. Sabouraudia. 1984;22(1):53–63. [PubMed] [Google Scholar]
  40. Rüchel R. On the renin-like activity of Candida proteinases and activation of blood coagulation in vitro. Zentralbl Bakteriol Mikrobiol Hyg A. 1983 Sep;255(2-3):368–379. [PubMed] [Google Scholar]
  41. Sehested M., Skovsgaard T., Jensen P. B., Demant E. J., Friche E., Bindslev N. Transport of the multidrug resistance modulators verapamil and azidopine in wild type and daunorubicin resistant Ehrlich ascites tumour cells. Br J Cancer. 1990 Jul;62(1):37–41. doi: 10.1038/bjc.1990.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sherman F. Getting started with yeast. Methods Enzymol. 1991;194:3–21. doi: 10.1016/0076-6879(91)94004-v. [DOI] [PubMed] [Google Scholar]
  43. Togni G., Sanglard D., Falchetto R., Monod M. Isolation and nucleotide sequence of the extracellular acid protease gene (ACP) from the yeast Candida tropicalis. FEBS Lett. 1991 Jul 29;286(1-2):181–185. doi: 10.1016/0014-5793(91)80969-a. [DOI] [PubMed] [Google Scholar]
  44. Vanden Bossche H., Marichal P., Gorrens J., Bellens D., Moereels H., Janssen P. A. Mutation in cytochrome P-450-dependent 14 alpha-demethylase results in decreased affinity for azole antifungals. Biochem Soc Trans. 1990 Feb;18(1):56–59. doi: 10.1042/bst0180056. [DOI] [PubMed] [Google Scholar]
  45. Vanden Bossche H., Marichal P., Odds F. C. Molecular mechanisms of drug resistance in fungi. Trends Microbiol. 1994 Oct;2(10):393–400. doi: 10.1016/0966-842x(94)90618-1. [DOI] [PubMed] [Google Scholar]
  46. Whelan W. L., Magee P. T. Natural heterozygosity in Candida albicans. J Bacteriol. 1981 Feb;145(2):896–903. doi: 10.1128/jb.145.2.896-903.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Wingard J. R., Merz W. G., Rinaldi M. G., Johnson T. R., Karp J. E., Saral R. Increase in Candida krusei infection among patients with bone marrow transplantation and neutropenia treated prophylactically with fluconazole. N Engl J Med. 1991 Oct 31;325(18):1274–1277. doi: 10.1056/NEJM199110313251803. [DOI] [PubMed] [Google Scholar]

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