Comparison between Conventional Cytogenetics and Interphase Fluorescence in situ Hybridization (FISH) for Patients with Multiple Myeloma

Sung-Hyun Kim; Jung Hwan Kim; Dong Mee Lee; Suee Lee; Sung Yong Oh; Hyuk-Chan Kwon; Kyung-Eun Kim; Jin-Yeong Han; Hyo-Jin Kim

doi:10.5045/kjh.2009.44.1.14

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Kim, Kim, Lee, Lee, Oh, Kwon, Kim, Han, and Kim: Comparison between Conventional Cytogenetics and Interphase Fluorescence in situ Hybridization (FISH) for Patients with Multiple Myeloma

Original Article

Korean J Hematol 2009;44(1):14-21.

Published online: 20 March 2009

DOI: https://doi.org/10.5045/kjh.2009.44.1.14

Comparison between Conventional Cytogenetics and Interphase Fluorescence in situ Hybridization (FISH) for Patients with Multiple Myeloma

Sung-Hyun Kim, M.D.¹, Jung Hwan Kim, M.D.¹, Dong Mee Lee, M.D.¹, Suee Lee, M.D.¹, Sung Yong Oh, M.D.¹, Hyuk-Chan Kwon, M.D.¹, Kyung-Eun Kim, M.D.², Jin-Yeong Han, M.D.², Hyo-Jin Kim, M.D.^1,

¹Department of Internal Medicine, Dong-A University College of Medicine, Busan, Korea

²Department 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

Received 9 August 2008 Revised 12 January 2009 Accepted 18 January 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.

Keywords: Multiple myeloma, Conventional cytogenetics, Fluorescence in situ hybridization

References

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Fig. 1.

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.

Fig. 2.

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).

Table 1.

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.

Table 2.

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.

Table 3.

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

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.

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