Abstract
We evaluated the expression of MDR1/p-glycoprotein in paediatric tumours using reverse transcriptase polymerase chain reaction (RT-PCR), RNA dot blot analysis, and immunohistochemistry on formalin fixed paraffin-embedded material with JSB-1 and C-219 monoclonal antibodies, and compared these three techniques. The expression of multidrug resistance-associated protein (MRP) gene was examined by RT-PCR assay. We studied MDR1/p-glycoprotein and MRP expression in 13 samples from 10 neuroblastoma patients, 11 samples from 10 nephroblastoma patients, 2 rhabdomyosarcomas, 1 adrenocortical carcinoma and 10 benign tumours or tumour-like lesions. Eleven of 13 neuroblastomas, 7 of 11 nephroblastomas, 2 rhabdomyosarcomas, 1 adrenocortical carcinoma, and 7 of 10 benign tumours or tumour-like lesions showed MDR1 PCR products. By RNA dot blot analysis, MDR1 transcripts were detectable in 11 of 34 specimens. Immunohistochemically, we detected positive reaction products for JSB-1 in 26 of 36 samples. There was a significant correlation between the immunoreactivity for JSB-1 and the expression of MDR1 mRNA expression by RTPCR (P=0.0001). However, the presence of p-glycoprotein immunostaining does not correlate with the MDR1 expression shown by RT-PCR in every case. As for MRP mRNA expression, 9 of 13 neuroblastomas and 10 of 11 nephroblastomas revealed PCR products.
Similar content being viewed by others
References
Akiyama S, Fojo A, Hanover JA, Pastan I, Gottesman MM (1985) Isolation and genetic characterization of human KB cell lines resistant to multiple drugs. Somatic Cell Genet 11:117–126
Barrand MA, Heppel-Parton AC, Wright KA, Rabbitts PH, Twentyman PR (1994) A 190-kilodalton protein overexpression in non-p-glycoprotein-containing multidrug-resistant cells and its relationship to the MRP gene. J Natl Cancer Int 86:110–117
Bates SE, Shieh CY, Tsokos M (1991) Expression of mdrl/P-glycoprotein in human neuroblastoma. Am J Pathol 139:305–315
Benard J, Bourhis J, Riou G (1991) Activation of N-myc oncogene and associated genes to chemoresistance, prognostic value for neuroblastoma. Bull Cancer (Paris) 78:91–97
Benard J, Bourhis J, de-Vathaire F, Ferrandis E, Terrier-Lacombe MJ, Lemerle J, Riou G, Hartmann O (1994) Prognostic value of MDR1 gene expression in neuroblastoma: results of a multivariante analysis. Prog Clin Biol Res 385:111–16
Bordow SB, Haber M, Madafiglio J, Cheung B, Marshall GM, Norris MD (1994) Expression of the multidrug resistance-associated protein (MRP) gene correlates with amplification and overexpression of the N-myc oncogene in childhood neuroblastoma. Cancer Res 54:5036–5040
Bourhis J, Benard J, Hartmann O, Boccon-Gibod L, Lemerle J, Riou G (1989) Correlation of MDR1 gene expression with chemotherapy in neuroblastoma. J Natl Cancer Inst 81:1401–1405
Brophy NA, Marie JP, Rojas VA, Warnke RA, McFall PJ, Smith SD, Sikic BI (1994) Mdr1 gene expression in childhood acute lymphoblastic leukemias and lymphomas: A critical evaluation by four techniques. Leukemia 8:327–335
Chan HSL, Haddad G, Thorner PS, DeBoer G, Lin YP, Ondrusek N, Yeger H, Ling V (1990) P-glycoprotein expression as a predictor of the outcome of therapy for neuroblastoma. N Engl J Med 325:1608–1614
Chen G, Jaffrezou J-P, Fleming WH, Duran GE, Sikic BI (1994) Prevalence of multidrug resistance related to activation of the mdrl gene in human sarcoma mutants derived by single-step doxorubicin selection. Cancer Res 54:4980–4987
Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159
Cole SPC, Bhardwaj G, Gerlach JH, Mackie JE, Grant CE, Almquist KC, Stewart AJ, Kurz EU, Duncan AMV, Deeley RG (1992) Overexpression of a transporter gene in a multidrug-resistant human lung cancer cell line. Science 258:1650–1654
Cordon-Cardo C, O'Brien JP, Boccia J, Casals D, Bertino Jr, Melamed MR (1990) Expression of the multidrug resistance gene product (p-glycoprotein) in human normal and tumor tissues. J Histochem Cytochem 38:1277–1287
Corrias MV, Cornaglia-Ferraris P, Di-Martino D, Stenger AM, Lanino E, Boni L, Tonini GP (1990) Expression of multiple drug resistance gene, MDR1, and N-myc oncogene in Italian population of human neuroblastoma patients. Anticancer Res 10:897–902
D'Angio GJ, Breslow N, Beckwith B, Evans A, Baum E, DeLimier A, Fernbach D, Hrabovsky E, Jones B, Kelalis P, Othersen B, Tefft M, Thomas PRM (1989) Treatment of Wilms' tumor: Results of the third national Wilms' tumor study. Cancer 64:349–360
Dietel M (1991) What's new in cytostatic drug resistance and pathology. Pathol Res Pract 187:892–905
Dietel M (1993) Meeting report. Second international symposium on cytostatic drug resistance. Cancer Res 53:2683–2688
Evans AE, D'Andio GJ, Randolph JA (1971) A proposed staging for children with neuroblastoma. Cancer 27:374–378
Fojo AT, Ueda K, Slamon DJ, Poplack DG, Gottesman MM, Pastan I (1987) Expression of a multidrug-resistance gene in human tumors and tissues. Proc Natl Acad Sci USA 84:265–269
Futscher BW, Abbaszadegan Mr, Domann F, Dalton WS (1994) Analysis of MRP mRNA in mitoxantrone-selected, multidrug-resistant human tumor cells. Biochem Pharmacol 47:1601–1606
Gekeler V, Beck J, Noller A, Wilisch A, Frese G, Neumann M, Handgretinger R, Ehninger G, Probst H, Niethammer D (1994) Drug-induced changes in the expression of MDR-associated genes: investigations on cultured cell lines and chemotherapeutically treated leukemias. Ann Hematol [Suppl] 69:519–524
Goldstein LJ, Galski H, Fojo A, Willingham M, Lai S-L, Gazdar A, Priker R, Green A, Crist W, Brodeur GM, Lieber M, Cossman J, Gottesman MM, Pastan I (1989) Expression of a multidrug resistance gene in human cancers. J Natl Cancer Inst 81:116–124
Goldstein LJ, Fojo AT, Ueda K, Crist W, Green A, Brodeur G, Pastan I, Gottesman MM (1990) Expression of the multidrug resistance, MDR1, gene in neuroblastoma. J Clin Oncol 8:128–136
Hughes M, Marsden HB, Palmer MK (1974) Histologic patterns of neuroblastoma related to prognosis and clinical staging. Cancer 34:1706–1711
Jereb B, Tournade MF, Lemerle J, Voute PA, Delemarre JF, Ahstrom L, Flamant R, Gerard-Marchant R, Sandstedt B (1980) Lymph node invasion and prognosis in nephroblastoma. Cancer 45:1632–1636
Juliano R, Ling A (1976) A surface glycoprotein modulation drug permeability in Chinese hamster ovarian cell mutants. Biochim Biophys Acta 455:152–162
Kandel RA, Campbell S, Noble-Topham S, Bell R, Andrulis IL (1995) Correlation of p-glycoprotein by immunohistochemistry with mdr-1 mRNA levels in osteosarcomas. Pilot study. Diagn Mol Pathol 4:59–65
Nakagawara A, Kadomatsu K, Sato S, Kohno K, Takano H, Akazawa K, Nose Y, Kuwano M (1990) Inverse correlation between expression of multidrug resistance gene and N-myc oncogene in human neuroblastoma. Cancer Res 50:3043–3047
Noonan K, Beck C, Holtzmayer T, Chin JE, Wunder JS, Andrulis IL, Gazdar AF, Willman CL, Griffith B, Von Hoff DD, Roninson IB (1990) Quantitative analysis of MDRI (multidrug resistance) gene expression in human tumors by polymerase chain reaction. Proc Natl Acad Sci USA 87:7160–7164
McDowell H, Peuchmaur M, Dominici C, Flamant F (1993) Multidrug resistance gene transcript level, and p-glycoprotein expression in paediatric malignant mesenchymal tumours. Anticancer Res 13:1863–1866
Pastan I, Gottesman M (1987) Multidrug resistance in human cancer. N Engl J Med 316:1388–1393
Ramani P, Dewchand H (1995) Expression of mdrl/P-glycoprotein and pl10 in neuroblastoma. J Pathol 175:13–22
Schmidt D, Harms D (1983) Histology and prognosis of nephroblastoma — with particular reference to special variants. Klin Padiatr 195:214–221
Shen DW, Fojo A, Chin JE, Roninson IB, Richert N, Pastan I, Gottesman MM (1986) Human multidrug-resistant cell lines: Increased mdrl expression can precede gene amplification. Science 232:643–645
Sugawara I, Arai T, Yamashita T, Yoshida A, Masunaga A, Itoyama S (1994) Expression of multidrug resistance-associated protein (MRP) in anaplastic carcinoma of the thyroid. Cancer Lett 82:185–188
Ueda K, Cornwell MM, Gottesman MM, Pastan I, Roninson IB, Ling V, Riordan JR (1986) The mdr1 gene, responsible for multidrug-resistance, codes for P-glycoprotein. Biochem Biophys Res Commun 141:956–962
Ueda K, Clark DP, Chen C, Roninson IB, Gottesman MM, Pastan I (1987) The human multidrug resistance (mdrl) gene: cDNA cloning and tanscription initiation. J Biol Chem 262:505–508
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Oda, Y., Röse, I., Radig, K. et al. Expression of MDR1/p-glycoprotein and multidrug resistance-associated protein in childhood solid tumours. Virchows Archiv 430, 99–105 (1997). https://doi.org/10.1007/BF01008030
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01008030