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Published in: BMC Urology 1/2012

Open Access 01-12-2012 | Research article

Determination of the differential expression of mitochondrial long non-coding RNAs as a noninvasive diagnosis of bladder cancer

Authors: Alexis Rivas, Verónica Burzio, Eduardo Landerer, Vincenzo Borgna, Sebastian Gatica, Rodolfo Ávila, Constanza López, Claudio Villota, Rodrigo de la Fuente, Javiera Echenique, Luis O Burzio, Jaime Villegas

Published in: BMC Urology | Issue 1/2012

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Abstract

Background

Bladder cancer is a significant cause of morbidity and mortality with a high recurrence rate. Early detection of bladder cancer is essential in order to remove the tumor, to preserve the organ and to avoid metastasis. The aim of this study was to analyze the differential expression of mitochondrial non-coding RNAs (sense and antisense) in cells isolated from voided urine of patients with bladder cancer as a noninvasive diagnostic assay.

Methods

The differential expression of the sense (SncmtRNA) and the antisense (ASncmtRNAs) transcripts in cells isolated from voided urine was determined by fluorescent in situ hybridization. The test uses a multiprobe mixture labeled with different fluorophores and takes about 1 hour to complete. We examined the expression of these transcripts in cells isolated from urine of 24 patients with bladder cancer and from 15 healthy donors.

Results

This study indicates that the SncmtRNA and the ASncmtRNAs are stable in cells present in urine. The test reveals that the expression pattern of the mitochondrial transcripts can discriminate between normal and tumor cells. The analysis of 24 urine samples from patients with bladder cancer revealed expression of the SncmtRNA and down-regulation of the ASncmtRNAs. Exfoliated cells recovered from the urine of healthy donors do not express these mitochondrial transcripts. This is the first report showing that the differential expression of these mitochondrial transcripts can detect tumor cells in the urine of patients with low and high grade bladder cancer.

Conclusion

This pilot study indicates that fluorescent in situ hybridization of cells from urine of patients with different grades of bladder cancer confirmed the tumor origin of these cells. Samples from the 24 patients with bladder cancer contain cells that express the SncmtRNA and down-regulate the ASncmtRNAs. In contrast, the hybridization of the few exfoliated cells recovered from healthy donors revealed no expression of these mitochondrial transcripts. This assay can be explored as a non-invasive diagnostic tool for bladder cancer.
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Literature
1.
go back to reference Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin. 2011, 61: 69-90. 10.3322/caac.20107.CrossRefPubMed Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer statistics. CA Cancer J Clin. 2011, 61: 69-90. 10.3322/caac.20107.CrossRefPubMed
2.
go back to reference Kaufman DS, Shipley WU, Feldman AS: Bladder cancer. Lancet. 2009, 374: 239-249. 10.1016/S0140-6736(09)60491-8.CrossRefPubMed Kaufman DS, Shipley WU, Feldman AS: Bladder cancer. Lancet. 2009, 374: 239-249. 10.1016/S0140-6736(09)60491-8.CrossRefPubMed
3.
go back to reference Van Tilborg AA, Bangma CH, Zwarthoff EC: Bladder cancer biomarkers and their role in surveillance and screening. Int J Urol. 2009, 16: 23-30. 10.1111/j.1442-2042.2008.02174.x.CrossRefPubMed Van Tilborg AA, Bangma CH, Zwarthoff EC: Bladder cancer biomarkers and their role in surveillance and screening. Int J Urol. 2009, 16: 23-30. 10.1111/j.1442-2042.2008.02174.x.CrossRefPubMed
4.
go back to reference Maier U, Simak R, Neuhold N: The clinical value of urinary cytology: 12 years of experience with 615 patients. J Clin Pathol. 1995, 48: 314-317. 10.1136/jcp.48.4.314.CrossRefPubMedPubMedCentral Maier U, Simak R, Neuhold N: The clinical value of urinary cytology: 12 years of experience with 615 patients. J Clin Pathol. 1995, 48: 314-317. 10.1136/jcp.48.4.314.CrossRefPubMedPubMedCentral
5.
go back to reference Ross J, Cohen M: Ancilliary methods for the detection of recurrent urothelial neoplasia. Cancer. 2000, 90: 75-86. 10.1002/(SICI)1097-0142(20000425)90:2<75::AID-CNCR2>3.0.CO;2-W.CrossRefPubMed Ross J, Cohen M: Ancilliary methods for the detection of recurrent urothelial neoplasia. Cancer. 2000, 90: 75-86. 10.1002/(SICI)1097-0142(20000425)90:2<75::AID-CNCR2>3.0.CO;2-W.CrossRefPubMed
6.
go back to reference Msaouel P, Koutsilieris M: Diagnostic value of circulating tumor cells detection in bladder and urothelial cancer: systematic review and meta-analysis. BMC Cancer. 2011, 11: 336-349. 10.1186/1471-2407-11-336.CrossRefPubMedPubMedCentral Msaouel P, Koutsilieris M: Diagnostic value of circulating tumor cells detection in bladder and urothelial cancer: systematic review and meta-analysis. BMC Cancer. 2011, 11: 336-349. 10.1186/1471-2407-11-336.CrossRefPubMedPubMedCentral
7.
go back to reference Ecke T: Focus on urinary bladder cancer markers: a review. Minerva Urol Nefrol. 2008, 60: 237-246.PubMed Ecke T: Focus on urinary bladder cancer markers: a review. Minerva Urol Nefrol. 2008, 60: 237-246.PubMed
8.
go back to reference Sokolova IA, Halling KC, Jenkins RB, Burkhardt HM, Meyer RG, Seelig SA, King W: The development of a multitarget, multicolor fluorescence in situ hybridization assay for the detection of urothelial carcinoma in urine. J Mol Diagn. 2000, 2: 116-123. 10.1016/S1525-1578(10)60625-3.CrossRefPubMedPubMedCentral Sokolova IA, Halling KC, Jenkins RB, Burkhardt HM, Meyer RG, Seelig SA, King W: The development of a multitarget, multicolor fluorescence in situ hybridization assay for the detection of urothelial carcinoma in urine. J Mol Diagn. 2000, 2: 116-123. 10.1016/S1525-1578(10)60625-3.CrossRefPubMedPubMedCentral
9.
go back to reference Villegas J, Burzio V, Villota C, Landerer E, Martinez R, Santander M, Martinez R, Pinto R, Vera MI, Boccardo E, Villa LL, Burzio LO: Expression of a novel non-coding mitochondrial RNA in human proliferating cells. Nucleic Acids Res. 2007, 35: 7336-7347. 10.1093/nar/gkm863.CrossRefPubMedPubMedCentral Villegas J, Burzio V, Villota C, Landerer E, Martinez R, Santander M, Martinez R, Pinto R, Vera MI, Boccardo E, Villa LL, Burzio LO: Expression of a novel non-coding mitochondrial RNA in human proliferating cells. Nucleic Acids Res. 2007, 35: 7336-7347. 10.1093/nar/gkm863.CrossRefPubMedPubMedCentral
10.
go back to reference Burzio VA, Villota C, Villegas J, Landerer E, Boccardo E, Villa LL, Martínez R, Lopez C, Gaete F, Toro V, Rodriguez X, Burzio LO: Expression of a family of noncoding mitochondrial RNAs distinguishes normal from cancer cells. Proc Natl Acad Sci USA. 2009, 106: 9430-9434. 10.1073/pnas.0903086106.CrossRefPubMedPubMedCentral Burzio VA, Villota C, Villegas J, Landerer E, Boccardo E, Villa LL, Martínez R, Lopez C, Gaete F, Toro V, Rodriguez X, Burzio LO: Expression of a family of noncoding mitochondrial RNAs distinguishes normal from cancer cells. Proc Natl Acad Sci USA. 2009, 106: 9430-9434. 10.1073/pnas.0903086106.CrossRefPubMedPubMedCentral
11.
go back to reference Villota C, Campos A, Vidaurre S, Oliveira-Cruz L, Boccardo E, Burzio VA, Varas M, Villegas J, Villa LL, Valenzuela PDT, Socias M, Roberts S, Burzio LO: Expression of mitocondrial ncRNAs is modulated by high risk HPV oncogenes. J Biol Chem. 2012, 287: 21303-21315. 10.1074/jbc.M111.326694.CrossRefPubMedPubMedCentral Villota C, Campos A, Vidaurre S, Oliveira-Cruz L, Boccardo E, Burzio VA, Varas M, Villegas J, Villa LL, Valenzuela PDT, Socias M, Roberts S, Burzio LO: Expression of mitocondrial ncRNAs is modulated by high risk HPV oncogenes. J Biol Chem. 2012, 287: 21303-21315. 10.1074/jbc.M111.326694.CrossRefPubMedPubMedCentral
12.
go back to reference Landerer E, Villegas J, Burzio VA, Oliveira L, Villota C, Lopez C, Restovic F, Martinez R, Castillo O, Burzio LO: Nuclear localization of the mitochondrial ncRNAs in normal and cancer cells. Cellular Oncol. 2011, 34: 297-305. 10.1007/s13402-011-0018-8.CrossRef Landerer E, Villegas J, Burzio VA, Oliveira L, Villota C, Lopez C, Restovic F, Martinez R, Castillo O, Burzio LO: Nuclear localization of the mitochondrial ncRNAs in normal and cancer cells. Cellular Oncol. 2011, 34: 297-305. 10.1007/s13402-011-0018-8.CrossRef
13.
go back to reference Dergunova NN, Bulycheva TI, Artemenko EG, Shpakova AP, Pegova AN, Gemjian EG, Dudnik OA, Zatsepina OV, Malashenko OS: A major nucleolar protein B23 as a marker of proliferation activity of human peripheral lymphocytes. Immunol Lett. 2002, 83: 67-72. 10.1016/S0165-2478(02)00085-8.CrossRefPubMed Dergunova NN, Bulycheva TI, Artemenko EG, Shpakova AP, Pegova AN, Gemjian EG, Dudnik OA, Zatsepina OV, Malashenko OS: A major nucleolar protein B23 as a marker of proliferation activity of human peripheral lymphocytes. Immunol Lett. 2002, 83: 67-72. 10.1016/S0165-2478(02)00085-8.CrossRefPubMed
14.
go back to reference Babjuk M, Oosterlinck W, Sylvester R, Kaasinen E, Böhle A, Palou-Redorta J, Morgan Roupret M: EAU guidelines on non–muscle-invasive urothelial carcinoma of the bladder, the 2011 update. Eur Urol. 2011, 59: 997-1008. 10.1016/j.eururo.2011.03.017.CrossRefPubMed Babjuk M, Oosterlinck W, Sylvester R, Kaasinen E, Böhle A, Palou-Redorta J, Morgan Roupret M: EAU guidelines on non–muscle-invasive urothelial carcinoma of the bladder, the 2011 update. Eur Urol. 2011, 59: 997-1008. 10.1016/j.eururo.2011.03.017.CrossRefPubMed
15.
go back to reference Wolff DJ, Bagg A, Cooley LD, Dewald GW, Hirsch BA, Jacky PB, Rao KW, Rao PN: Guidance for fluorescencein situhybridization testing in hematologic disorders. J Mol Diagn. 2007, 9: 134-143. 10.2353/jmoldx.2007.060128.CrossRefPubMedPubMedCentral Wolff DJ, Bagg A, Cooley LD, Dewald GW, Hirsch BA, Jacky PB, Rao KW, Rao PN: Guidance for fluorescencein situhybridization testing in hematologic disorders. J Mol Diagn. 2007, 9: 134-143. 10.2353/jmoldx.2007.060128.CrossRefPubMedPubMedCentral
16.
go back to reference Halling KC, King W, Sokolova IA, Meyer RG, Burkhardt HM, Halling AC, Cheville JC, Sebo TJ, Ramakumar S, Stewart CS, Pankratz S, O’Kane DJ, Seelig SA, Lieber MM, Jenkins RB: A comparison of cytology and fluorescencein situhybridization for the detection of urothelial carcinoma. J Urol. 2000, 164: 1768-1775. 10.1016/S0022-5347(05)67104-2.CrossRefPubMed Halling KC, King W, Sokolova IA, Meyer RG, Burkhardt HM, Halling AC, Cheville JC, Sebo TJ, Ramakumar S, Stewart CS, Pankratz S, O’Kane DJ, Seelig SA, Lieber MM, Jenkins RB: A comparison of cytology and fluorescencein situhybridization for the detection of urothelial carcinoma. J Urol. 2000, 164: 1768-1775. 10.1016/S0022-5347(05)67104-2.CrossRefPubMed
17.
go back to reference Kang JU, Koo SH, Jeong TE, Kwon KC, Park JW, Jeon CH: Multitarget fluorescencein situhybridization and melanoma antigen genes analysis in primary bladder carcinoma. Cancer Genet Cytogenet. 2006, 164: 32-38. 10.1016/j.cancergencyto.2005.06.006.CrossRefPubMed Kang JU, Koo SH, Jeong TE, Kwon KC, Park JW, Jeon CH: Multitarget fluorescencein situhybridization and melanoma antigen genes analysis in primary bladder carcinoma. Cancer Genet Cytogenet. 2006, 164: 32-38. 10.1016/j.cancergencyto.2005.06.006.CrossRefPubMed
18.
go back to reference Kipp BR, Tanasescu M, Else TA, Bryant SC, Karnes RJ, Sebo TJ, Halling KC: Quantitative fluorescencein situhybridization and its ability to predict bladder cancer recurrence and progression to muscle-invasive bladder cancer. J Mol Diagn. 2009, 11: 148-154. 10.2353/jmoldx.2009.080096.CrossRefPubMedPubMedCentral Kipp BR, Tanasescu M, Else TA, Bryant SC, Karnes RJ, Sebo TJ, Halling KC: Quantitative fluorescencein situhybridization and its ability to predict bladder cancer recurrence and progression to muscle-invasive bladder cancer. J Mol Diagn. 2009, 11: 148-154. 10.2353/jmoldx.2009.080096.CrossRefPubMedPubMedCentral
19.
go back to reference Colucci G, Floege J, Schena FP: The urinary sediment beyond light microscopical examination. Nephrol Dial Transplant. 2006, 21: 1482-1485. 10.1093/ndt/gfl223.CrossRefPubMed Colucci G, Floege J, Schena FP: The urinary sediment beyond light microscopical examination. Nephrol Dial Transplant. 2006, 21: 1482-1485. 10.1093/ndt/gfl223.CrossRefPubMed
20.
go back to reference Meiers I, Singh H, Hossain D, Lang K, Liu L, Qian J, Verhest AP, Bostwick DG: Improved filter method for urine sediment detection of urothelial carcinoma by fluorescencein situhybridization. Arch Pathol Lab Med. 2007, 131: 1574-1579.PubMed Meiers I, Singh H, Hossain D, Lang K, Liu L, Qian J, Verhest AP, Bostwick DG: Improved filter method for urine sediment detection of urothelial carcinoma by fluorescencein situhybridization. Arch Pathol Lab Med. 2007, 131: 1574-1579.PubMed
21.
go back to reference Stewart AK, Fonseca R: Review of molecular diagnostics in multiple myeloma. Expert Rev Mol Diagn. 2007, 7: 453-459. 10.1586/14737159.7.4.453.CrossRefPubMed Stewart AK, Fonseca R: Review of molecular diagnostics in multiple myeloma. Expert Rev Mol Diagn. 2007, 7: 453-459. 10.1586/14737159.7.4.453.CrossRefPubMed
22.
go back to reference Bryce AH, Ketterling RP, Gertz MA, Lacy M, Knudson RA, Zeldenrust S, Kumar S, Hayman S, Buadi F, Kyle RA, Greipp PR, Lust JA, Russell S, Rajkumar SV, Fonseca R, Dispenzieri A: Translocation t(11;14) and survival of patients with light chain (AL) amyloidosis. Haematologica. 2009, 94: 380-386. 10.3324/haematol.13369.CrossRefPubMedPubMedCentral Bryce AH, Ketterling RP, Gertz MA, Lacy M, Knudson RA, Zeldenrust S, Kumar S, Hayman S, Buadi F, Kyle RA, Greipp PR, Lust JA, Russell S, Rajkumar SV, Fonseca R, Dispenzieri A: Translocation t(11;14) and survival of patients with light chain (AL) amyloidosis. Haematologica. 2009, 94: 380-386. 10.3324/haematol.13369.CrossRefPubMedPubMedCentral
23.
go back to reference Avet-Loiseau H, Soulier J, Fermand JP, Yakoub-Agh : IFM and MAG groups. Impact of high-risk cytogenetics and prior therapy on outcomes in patients with advanced relapsed or refractory multiple myeloma treated with lenalidomide plus dexaméthasone. Leukemia. 2010, 24: 623-628. 10.1038/leu.2009.273.CrossRefPubMed Avet-Loiseau H, Soulier J, Fermand JP, Yakoub-Agh : IFM and MAG groups. Impact of high-risk cytogenetics and prior therapy on outcomes in patients with advanced relapsed or refractory multiple myeloma treated with lenalidomide plus dexaméthasone. Leukemia. 2010, 24: 623-628. 10.1038/leu.2009.273.CrossRefPubMed
24.
go back to reference Pauletti G, Godolphin W, Press MF, Slamon DJ: Detection and quantitation of HER-2/neu gene amplification in human breast cancer archival material using fluorescencein situhybridization. Oncogene. 1996, 13: 63-72.PubMed Pauletti G, Godolphin W, Press MF, Slamon DJ: Detection and quantitation of HER-2/neu gene amplification in human breast cancer archival material using fluorescencein situhybridization. Oncogene. 1996, 13: 63-72.PubMed
25.
go back to reference Escudier SM, Pereira-Leahy JM, Drach JW, Weier HU, Goodacre AM, Cork MA, Trujillo JM, Keating MJ, Andreeff M: Fluorescentin situhybridization and cytogenetic studies of trisomy 12 in chronic lymphocytic leukemia. Blood. 1993, 81: 2702-2707.PubMed Escudier SM, Pereira-Leahy JM, Drach JW, Weier HU, Goodacre AM, Cork MA, Trujillo JM, Keating MJ, Andreeff M: Fluorescentin situhybridization and cytogenetic studies of trisomy 12 in chronic lymphocytic leukemia. Blood. 1993, 81: 2702-2707.PubMed
26.
go back to reference Hagemeijer A, Buijs A, Smit E, Janssen B, Creemers GJ, Van der Plas D, Grosveld G: Translocation of BCR to chromosome 9: a new cytogenetic variant detected by FISH in two Ph-negative, BCR-positive patients with chronic myeloid leukemia. Genes Chromosomes Cancer. 1993, 8: 237-245. 10.1002/gcc.2870080406.CrossRefPubMed Hagemeijer A, Buijs A, Smit E, Janssen B, Creemers GJ, Van der Plas D, Grosveld G: Translocation of BCR to chromosome 9: a new cytogenetic variant detected by FISH in two Ph-negative, BCR-positive patients with chronic myeloid leukemia. Genes Chromosomes Cancer. 1993, 8: 237-245. 10.1002/gcc.2870080406.CrossRefPubMed
27.
go back to reference Zanardi A, Bandiera D, Bertolini F, Corsini CA, Gregato G, Milani P, Barborini E, Carbone R: Miniaturized FISH for screening of onco-hematological malignancies. Biotechniques. 2010, 49: 497-504. 10.2144/000113445.CrossRefPubMed Zanardi A, Bandiera D, Bertolini F, Corsini CA, Gregato G, Milani P, Barborini E, Carbone R: Miniaturized FISH for screening of onco-hematological malignancies. Biotechniques. 2010, 49: 497-504. 10.2144/000113445.CrossRefPubMed
28.
go back to reference Nelson B, Gupta R, Dewald G, Paternoster S, Rosen S, Peterson L: Chronic lymphocytic leukemia FISH impact on diagnosis panel. American J Clinical Pathol. 2007, 128: 323-332. 10.1309/21TN2RUWKR827UW2.CrossRef Nelson B, Gupta R, Dewald G, Paternoster S, Rosen S, Peterson L: Chronic lymphocytic leukemia FISH impact on diagnosis panel. American J Clinical Pathol. 2007, 128: 323-332. 10.1309/21TN2RUWKR827UW2.CrossRef
29.
go back to reference Jiang F, Caraway NP, Nebiyou Bekele B, Zhang HZ, Khanna A, Wang H, Li R, Fernandez RL, Zaidi TM, Johnston DA, Katz RL: Surfactant protein A gene deletion and prognostics for patients with stage I non-small cell lung cancer. Clin Cancer Res. 2005, 11: 5417-5424. 10.1158/1078-0432.CCR-04-2087.CrossRefPubMed Jiang F, Caraway NP, Nebiyou Bekele B, Zhang HZ, Khanna A, Wang H, Li R, Fernandez RL, Zaidi TM, Johnston DA, Katz RL: Surfactant protein A gene deletion and prognostics for patients with stage I non-small cell lung cancer. Clin Cancer Res. 2005, 11: 5417-5424. 10.1158/1078-0432.CCR-04-2087.CrossRefPubMed
30.
go back to reference Li R, Liu Z, Fan T, Jiang F: A novel multiple FISH array for the detection of genetic aberrations in cancer. Lab Invest. 2006, 86: 619-627.CrossRefPubMed Li R, Liu Z, Fan T, Jiang F: A novel multiple FISH array for the detection of genetic aberrations in cancer. Lab Invest. 2006, 86: 619-627.CrossRefPubMed
Metadata
Title
Determination of the differential expression of mitochondrial long non-coding RNAs as a noninvasive diagnosis of bladder cancer
Authors
Alexis Rivas
Verónica Burzio
Eduardo Landerer
Vincenzo Borgna
Sebastian Gatica
Rodolfo Ávila
Constanza López
Claudio Villota
Rodrigo de la Fuente
Javiera Echenique
Luis O Burzio
Jaime Villegas
Publication date
01-12-2012
Publisher
BioMed Central
Published in
BMC Urology / Issue 1/2012
Electronic ISSN: 1471-2490
DOI
https://doi.org/10.1186/1471-2490-12-37

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