Expression level of MDR1 message in peripheral blood leukocytes from healthy adults: a competitive nucleic acid sequence-based amplification assay for its determination
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Hiroyuki Kobayashi
, Yuzuru Takemura , Tsukasa Hayashi , Takeshi Ujiiye , Masako Kawase , Yasuhiro Niino , Hayato Miyachi , Toshio Ohshima and Tomomitsu Hotta
Abstract
Accurate quantification of multidrug resistance-1 gene (MDR1) expression in target cells would be of important therapeutic value in predicting cellular response to anticancer drugs. Because certain normal cells in peripheral blood physiologically express MDR1, increasing the sensitivity of the detection methods might result in confounding low-degree expression in tumor cells with physiologic expression in normal cells. The purpose of this study was to determine MDR1 mRNA expression levels in peripheral blood leukocytes obtained from healthy adult volunteers using a competitive nucleic acid sequence-based amplification (NASBA) assay. We determined the reference intervals of MDR1 mRNA expression in peripheral blood obtained from 98 healthy adults by measuring its expression with the quantitative NASBA assay between 5.50 × 104 copies/µg RNA and 6.76 × 105 copies/µg RNA. The new reference intervals were evaluated using a number of sensitive or resistant cell lines as control; positive or negative MDR1 expression was clearly demonstrated. We also reevaluated MDR1 expression levels in leukemia cells obtained from patient peripheral blood; 18 of 31 samples (58%) exceeded the newly established upper reference limit. The cutoff value established could be used to distinguish significant MDR1 expression in tumor cells from physiologic expression in certain normal cells coexistent in peripheral blood.
References
1 Ambudkar SV, Kimchi-Sarfaty C, Sauna ZE, Gottesman MM. P-glycoprotein: from genomics to mechanism. Oncogene 2003; 22:7468–85.10.1038/sj.onc.1206948Search in Google Scholar
2 Kim RB. Drug transporters in HIV therapy. Top HIV Med 2003; 11:136–9.Search in Google Scholar
3 Fujimaki S, Funato T, Harigae H, Fujiwara J, Kameoka J, Meguro K, et al. Quantitative analysis of a MDR1 transcript for prediction of drug resistance in acute leukemia. Clin Chem 2002; 48:811–7.10.1093/clinchem/48.6.811Search in Google Scholar
4 Debuire B, Sol O, Lemoine A, May E. Nonisotopic competitive RT-PCR assay to measure MDR1 gene expression. Clin Chem 1995; 41:819–25.10.1093/clinchem/41.6.819Search in Google Scholar
5 van Hille B, Lohri A, Reuter J, Herrmann R. Nonradioactive quantification of mdr1 mRNA by polymerase chain reaction amplification coupled with HPLC. Clin Chem 1995; 41:1087–93.10.1093/clinchem/41.8.1087Search in Google Scholar
6 Beck WT, Grogan TM, Willman CL, Cordon-Cardo C, Parham DM, Kuttesch JF, et al. Methods to detect P-glycoprotein-associated multidrug resistance in patients’ tumors: consensus recommendations. Cancer Res 1996; 56:3010–20.Search in Google Scholar
7 Kobayashi H, Takemura Y, Kawai Y, Miyachi H, Kawabata M, Matsumura T, et al. Competitive reverse transcription-polymerase chain reaction assay for quantification of human multidrug resistance 1 (MDR1) gene expression in fresh leukemic cells. J Lab Clin Med 2000; 135:199–209.10.1067/mlc.2000.104461Search in Google Scholar
8 Klimecki WT, Futscher BW, Grogan TM, Dalton WS. P-glycoprotein expression and function in circulating blood cells from normal volunteers. Blood 1994; 83:2451–8.10.1182/blood.V83.9.2451.2451Search in Google Scholar
9 Chaudhary PM, Mechetner EB, Roninson IB. Expression and activity of the multidrug resistance P-glycoprotein in human peripheral blood lymphocytes. Blood 1992; 80:2735–9.10.1182/blood.V80.11.2735.bloodjournal80112735Search in Google Scholar
10 Hayashi T, Kobayashi H, Miyachi H, Ohshima T, Ujiiye T, Kawase M, et al. A competitive nucleic acid sequence-based amplification assay for the quantification of human MDR1 transcript in leukemia cells. Clin Chim Acta 2004; 342:115–26.10.1016/j.cccn.2003.12.013Search in Google Scholar
11 Kobayashi H, Takemura Y, Miyachi H, Kawabata M, Mori S, Kawai Y, et al. Quantitative analysis of human multidrug resistance 1 (MDR1) gene expression by nonisotopic competitive reverse transcriptase polymerase chain reaction assay. J Clin Lab Anal 1997; 11:258–66.10.1002/(SICI)1098-2825(1997)11:5<258::AID-JCLA4>3.0.CO;2-3Search in Google Scholar
12 Arkin H, Ohnuma T, Kamen BA, Holland JF, Vallabhajosula S. Multidrug resistance in a human leukemic cell line selected for resistance to trimetrexate. Cancer Res 1989; 49:6556–61.Search in Google Scholar
13 Tsuruo T, Iida H, Tsukagoshi S, Sakurai Y. Potentiation of vincristine and adriamycin effects in human hemopoietic tumor cell lines by calcium antagonists and calmodulin inhibitors. Cancer Res 1983; 43:2267–72.Search in Google Scholar
14 Tsuruo T, Iida-Saito H, Kawabata H, Oh-hara T, Hamada H, Utakoji T. Characteristics of resistance to adriamycin in human myelogenous leukemia K562 resistant to adriamycin and in isolated clones. Jpn J Cancer Res 1986; 77:682–92.Search in Google Scholar
15 Ralph P, Moore MA, Nilsson K. Lysozyme synthesis by established human and murine histiocytic lymphoma cell lines. J Exp Med 1976; 143:1528–33.10.1084/jem.143.6.1528Search in Google Scholar
16 Yokota J, Akiyama T, Fung YK, Benedict WF, Namba Y, Hanaoka M, et al. Altered expression of the retinoblastoma (RB) gene in small-cell carcinoma of the lung. Oncogene 1988; 3:471–5.Search in Google Scholar
17 Kyoizumi S, Akiyama M, Kouno N, Kobuke K, Hakoda M, Jones SL, et al. Monoclonal antibodies to human squamous cell carcinoma of the lung and their application to tumor diagnosis. Cancer Res 1985; 45:3274–81.Search in Google Scholar
18 Solberg HE. Establishment and use of reference values. In: Burtis CA, Ashwood ER, editors. Tietz textbook of clinical chemistry, 3rd ed. Philadelphia: WB Saunders, 1999:336–56.Search in Google Scholar
19 NCCLS. How to define and determine reference intervals in the clinical laboratory; approved guideline, 2nd ed. (NCCLS document C28-A2). Wayne: NCCLS, 2000.Search in Google Scholar
20 Leith CP, Kopecky KJ, Chen IM, Eijdems L, Slovak ML, McConnell TS, et al. Frequency and clinical significance of the expression of the multidrug resistance proteins MDR1/P-glycoprotein, MRP1, and LRP in acute myeloid leukemia: a Southwest Oncology Group Study. Blood 1999; 94:1086–99.Search in Google Scholar
21 Han K, Kahng J, Kim M, Lim J, Kim Y, Cho B, et al. Expression of functional markers in acute nonlymphoblastic leukemia. Acta Haematol 2000; 104:174–80.10.1159/000046511Search in Google Scholar PubMed
22 Tafuri A, Gregorj C, Petrucci MT, Ricciardi MR, Mancini M, Cimino G, et al. MDR1 protein expression is an independent predictor of complete remission in newly diagnosed adult acute lymphoblastic leukemia. Blood 2002; 100:974–81.10.1182/blood-2001-12-0371Search in Google Scholar PubMed
23 Broxterman HJ, Sonneveld P, van Putten WJ, Lankelma J, Eekman CA, Ossenkoppele GJ, et al. P-glycoprotein in primary acute myeloid leukemia and treatment outcome of idarubicin/cytosine arabinoside-based induction therapy. Leukemia 2000; 14:1018–24.10.1038/sj.leu.2401796Search in Google Scholar PubMed
24 List AF, Kopecky KJ, Willman CL, Head DR, Persons DL, Slovak ML, et al. Benefit of cyclosporine modulation of drug resistance in patients with poor-risk acute myeloid leukemia: a Southwest Oncology Group study. Blood 2001; 98:3212–20.10.1182/blood.V98.12.3212Search in Google Scholar
© Walter de Gruyter
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Articles in the same Issue
- The significance of serum γ-glutamyltransferase in cardiovascular diseases
- Anti-tissue transglutaminase antibodies in inflammatory bowel disease: new evidence
- Expression level of MDR1 message in peripheral blood leukocytes from healthy adults: a competitive nucleic acid sequence-based amplification assay for its determination
- Characteristics of prolinase against various iminodipeptides in erythrocyte lysates from a normal human and a patient with prolidase deficiency
- Alteration of homocysteine catabolism in pre-eclampsia, HELLP syndrome and placental insufficiency
- Immune system-mediated endothelial damage is associated with NO and antioxidant system disorders
- Bone marrow microvascular density and angiogenic growth factors in multiple myeloma
- The insulin-like growth factor system in the circulation of patients with viral infections
- Time course of systemic markers of inflammation in patients presenting with acute coronary syndromes
- Age- and sex-related reference values for serum insulin concentration and its biological determinants in a French healthy population. The STANISLAS cohort
- Potential pitfalls of comparative measurements of reticulocytes with flow cytometry and microscopy in prematures and infants
- Screening of antinuclear antibodies: comparison between enzyme immunoassay based on nuclear homogenates, purified or recombinant antigens and immunofluorescence assay
- Analytical and diagnostic accuracy of the EliA™ automated enzyme fluoroimmunoassay for antineutrophil cytoplasmic autoantibody detection
- A particle-enhanced turbidimetric immunoassay for quantitative determination of orosomucoid in urine: development, validation and reference values
- Analytical performance and clinical results of a fully automated MEIA system for brain natriuretic peptide assay: comparison with a point of care testing method
- Multicentre performance evaluation of the E170 Module for MODULAR ANALYTICS
- Congress of Clinical Chemistry and Laboratory Medicine Annual meeting of the German United Society for Clinical Chemistry and Laboratory Medicine (DGKL), Düsseldorf, Germany, November 22-24, 2004