kjh The Korean Journal of Hematology Korean J Hematol 1738-7949 2092-9129 Korean Society of Hematology 10.5045/kjh.2009.44.1.14 kjh-44-14 Original Article Comparison between Conventional Cytogenetics and Interphase Fluorescence in situ Hybridization (FISH) for Patients with Multiple Myeloma KimSung-HyunM.D. 1 KimJung HwanM.D. 1 LeeDong MeeM.D. 1 LeeSueeM.D. 1 OhSung YongM.D. 1 KwonHyuk-ChanM.D. 1 KimKyung-EunM.D. 2 HanJin-YeongM.D. 2 KimHyo-JinM.D. 1 Department of Internal Medicine, Dong-A University College of Medicine, Busan, Korea Department of Laboratory Medicine, Dong-A University College of Medicine, Busan, Korea Correspondence to：Hyo-Jin Kim, M.D., Ph.D. Department of Internal Medicine, Dong-A University College of Medicine 3-1, Dongdaeshin-dong, Seo-gu, Busan 602-715, Korea Tel: ＋82-51-240-2951, Fax: ＋82-51-240-2088 E-mail: kimhj@dau.ac.kr 032009 20032009 441 1421 09082008 12012009 18012009 Copyright © 2009 Korean Society of Hematology 2009 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Background

For patients with multiple myeloma (MM), different strategies are used to detect chromosomal abnormalities (CA). There have been a few studies that have directly compared FISH with conventional cytogenetics (CC) for the detection of CA. In this study, we employed a combined approach of metaphase cytogenetics and interphase FISH to investigate the genetic basis for the great heterogeneity observed in the clinical behavior of 28 MM patients.

 Methods

Cytogenetic analysis was performed via traditional metaphase karyotype analysis. The FISH studies were done using DNA probes to detect translocations involving the immunoglobulin heavy chain gene (IGH) at 14q32 and deletions of 17p13.1 and 13q14.

 Results

CA were detected by CC in 16 patients (57.1%) and by FISH in 14 patients (50.0%) of the 28 patients we studied. 14q32 abnormalities and deletion abnormalities of 13q14 and 17p13.1 were detected by CC in five patients (17.9%), three patients (10.7%) and no patients (0%), respectively and these were detected by FISH in 12 (42.8%), four (14.3%) and five (17.8%), respectively, of the 28 patients we studied. The median follow-up timefor the patients was 23.85 months (range: 0.3∼58.13 months). On the univariate and multivariate analyses, none of the abnormalities detected by cytogenetics and interphase FISH affected survival.

 Conclusion

On comparing the cytogenetics and interphase FISH results, we can suggest that both studies should be an essential part of the workup for the diagnosis of patients with MM. Also, both studies may complement each other to predict the prognosis.

 Multiple myeloma Conventional cytogenetics Fluorescence in situ hybridization References Bataille R Harousseau JL Multiple myeloma N Engl J Med 1997336 165764 Seong C Delasalle K Hayes K Prognostic value of cytogenetics in multiple myeloma Br J Haematol 1998101 18994 Bergsagel PL Kuehl WM Chromosome translocations in multiple myeloma Oncogene 200120 561122 Kaufmann H Krömer E Nösslinger T Both chromosome 13 abnormalities by metaphase cytogenetics and deletion of 13q by interphase FISH only are prognostically relevant in multiple myeloma Eur J Haematol 200371 17983 Fonseca R Blood E Rue M Clinical and biologic implications of recurrent genomic aberrations in myeloma Blood 2003101 456975 Drach J Ackermann J Fritz E Presence of a p53 gene deletion in patients with multiple myeloma predicts for short survival after conventional-dose chemotherapy Blood 199892 8029 Sawyer JR Waldron JA Jagannath S Barlogie B Cytogenetic findings in 200 patients with multiple myeloma Cancer Genet Cytogenet 199582 419 Chang H Li D Zhuang L Detection of chromosome 13q deletions and IgH translocations in patients with multiple myeloma by FISH: comparison with karyotype analysis Leuk Lymphoma 200445 9659 Dewald GW Therneau T Larson D Relationship of patient survival and chromosome anomalies detected in metaphase and/or interphase cells at diagnosis of myeloma Blood 2005106 35538 Gertz MA Lacy MQ Dispenzieri A Clinical implications of t(11;14)(q13;q32), t(4;14)(p16.3;q32), and −17p13 in myeloma patients treated with high-dose therapy Blood 2005106 283740 Shaughnessy J Jr Tian E Sawyer J Prognostic impact of cytogenetic and interphase fluorescence in situ hybridization-defined chromosome 13 deletion in multiple myeloma: early results of total therapy II Br J Haematol 2003120 4452 American College of Medical Genetics Standards and guidelines for clinical genetics laboratories 2006 Edition. Shaffer L Tommerup N An international system for human cytogenetic nomenclature. Basel, Switzerland: S. Karger, 2005. and haemopoietic stem cell transplantation. Myeloma Subcommittee of the EBMT. European Group for Blood and Marrow Transplant Br J Haematol 1998102 111523 BladéJ, Samson D, Reece D, et al Criteria for evaluating disease response and progression in patients with multiple myeloma treated by high-dose therapy Lai JL Zandecki M Mary JY Improved cytogenetics in multiple myeloma: a study of 151 patients including 117 patients at diagnosis Blood 199585 24907 Tabernero D San Miguel JF Garcia-Sanz M Incidence of chromosome numerical changes in multiple myeloma: fluorescence in situ hybridization analysis using 15 chromosome-specific probes Am J Pathol 1996149 15361 Drach J Schuster J Nowotny H Multiple myeloma: high incidence of chromosomal aneuploidy as detected by interphase fluorescence in situ hybridization Cancer Res 199555 38549 Nishida K Tamura A Nakazawa N The Ig heavy chain gene is frequently involved in chromosomal translocations in multiple myeloma and plasma cell leukemia as detected by in situ hybridization Blood 199790 52634 Fonseca R Barlogie B Bataille R Genetics and cytogenetics of multiple myeloma: a workshop report Cancer Res 200464 154658 Zojer N Königsberg R Ackermann J Deletion of 13q14 remains an independent adverse prognostic variable in multiple myeloma despite its frequent detection by interphase fluorescence in situ hybridization Blood 200095 192530 Avet-Loiseau H Li JY Godon C P53 deletion is not a frequent event in multiple myeloma Br J Haematol 1999106 7179 Schultheis B Krämer A Willer A Hegenbart U Goldschmidt H Hehlmann R Analysis of p73 and p53 gene deletions in multiple myeloma Leukemia 199913 2099103 Mazars GR Portier M Zhang XG Mutations of the p53 gene in human myeloma cell lines Oncogene 19927 10158 Chang H Qi C Yi QL Reece D Stewart AK p53 gene deletion detected by fluorescence in situ hybridization is an adverse prognostic factor for patients with multiple myeloma following autologous stem cell transplantation Blood 2005105 35860 Chiecchio L Protheroe RK Ibrahim AH Deletion of chromosome 13 detected by conventional cytogenetics is a critical prognostic factor in myeloma Leukemia 200620 16107 Figures and Tables

Interphase fluorescence in situ hybridization (FISH) findings. (A) A plasma cell shows rearrangement of IGH. (B) A plasma cell shows deletion 13q14. The locus-specific 13q14 probes are labeled by orange color. (C) A plasma cell shows deletion 17p13.1. The locus-specific p53 probes are labeled by orange color.

The overall survival curves according to the initial treatment response. For patients with a CR or a PR, the median survival has not yet been reached. The median survival was 7.03 months for patients with MR. There were significant differences in survival according to the initial treatment response (P=0.036). (CR; complete response, PR; partial response, MR; minimal response).

Characteristics of the 28 study patients

Characteristics n=28
Median age (range) 62.5 (44∼78)
Gender (male/female) 19/9
Durie-salmon stage (%) Stage I, II 0
Stage III 28 (100)
Median Cr, mg/Dl (range) 1.5 (0.7∼10.1)
Median beta2, ug/mL (range) 5.87 (1.53∼79.32)
Median LDH, IU/L (range) 348.5 (137∼881)
Median BM plasma cells, % (range) 37.1 (10∼91)
Median CRP, mg/dL (range) 1.24 (0.06∼15.3)
Median albumin, g/dL (range) 3.7 (2.6∼4.9)
Median serum M protein, g/dL (range) 3.65 (0∼7.05)
Median calcium, mg/dL (range) 10.2 (7.1∼15.4)
Median hemoglobin, g/dL (range) Immunoglobulin subtype (%) 9.2 (3.9∼13.4)
IgG 14 (50.0)
IgA 4 (14.3)
IgD 1 (3.6)
light chain disease 9 (32.1)
κ 16 (57.1)
λ 12 (42.9)

Abbreviations: Cr, creatinine; beta2, β2 microglobulin; LDH, lactic dehydrogenase; BM, bone marrow; CRP, C-reactive protein.

CC and FISH data from 28 MM patients

Karyotype Patients n Incidence in MM patients % FISH Patients n Incidence in MM patients %
AA 16 57.1 Detected 14 50
NA only 1 3.6 IGH only 7 25
SA only 3 10.7 del(13q) only 1 3.6
BA 12 42.8 del(17p) only 1 3.6
Any NA 13 46.4      
Hyperdiploidy 10 35.7 IGH and del(13q) 1 3.6
Hypodiploidy 2 7.1 IGH and del(17p) 2 7.1
Tetraploidy 1 3.6      
Any SA 15 53.6 del(13q) and del(17p) 0 0
14q32 − 5 − 17.9    
–13 or 13q14− − 3 10.7    
–17 or 17p− 0 0 IGH, del(13q) and del(17p) 2 7.1
      Any IGH 12 42.8
      Any del(13q) 4 14.3
      Any del(17p) 5 17.8

Abbreviations: CC, conventional cytogenetics; FISH, fluorescence in situ hybridization; MM, Multiple myeloma; IGH, immunoglobulin heavy chain gene; AA, any abnormalities; NA, numeric abnormalities; SA, structural abnormalities; BA, both numeric and structural abnormalities.

P value for the association between the bioclinical features and the abnormalities n=28

  Metaphase cytogenetics Interphase FISH
  CA IGH del(13q) del(17p)
Cr (mg/dL)
　≥2 0.006 0.119 0.357 0.018
　＜2
Beta 2 (ug/mL)
　≥3.5 0.012 0.529 0.465 0.256
　＜3.5
BM plasma cell (%)
　≥33 0.003 0.012 0.511 0.119
　＜33
M-protein (g/dL)
　≥5 0.024 0.939 0.158 0.298
　＜5
Ca (mg/dL)
　≥12 0.157 0.267 0.014 0.086
　＜12
Hb (g/dL)
　≥10 0.033 0.515 0.107 0.787
　＜10

Abbreviations: FISH, fluorescence in situ hybridization; CA, chromosomal abnormalities; IGH, immunoglobulin heavy chain gene; Cr, creatinine; beta2, β2 microglobulin; BM, bone marrow; Ca, calcium; Hb, hemoglobin.