Effect of Bone Decalcification Procedures on DNA In Situ Hybridization and Comparative Genomic Hybridization: EDTA Is Highly Preferable to a Routinely Used Acid Decalcifier

Alers, Janneke C.; Krijtenburg, Pieter-Jaap; Vissers, Kees J.; Dekken, Herman van

doi:10.1177/002215549904700512

Cited by 130 publications

(72 citation statements)

References 16 publications

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“…All of formalin-fixed, paraffin-embedded specimens received acidic pretreatment to decalcify bones, which is known to affect DNA and mRNA integrity. 32 EDTA decalcification, 52 in particular if combined with ultrasonic decalcification, 53 might increase the quality of the DNA extracted. However, the duration of such a decalcification protocol might make it difficult to implement in routine, apart from implementing in specialized centers.…”

Section: Discussionmentioning

confidence: 99%

Diagnostic value of investigating GNAS mutations in fibro-osseous lesions: a retrospective study of 91 cases of fibrous dysplasia and 40 other fibro-osseous lesions

et al. 2013

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nucleotide-binding protein/a-subunit) mutations that induce the activation of G-protein a-subunit participate in the pathogenesis of fibrous dysplasia. The aim of this study was to evaluate the sensitivity and specificity of GNAS mutations in fibrous dysplasia and other fibro-osseous lesions, to assess the value of investigating this mutation in the diagnosis of fibro-osseous lesions. We studied 91 cases of fibrous dysplasia. The quality and/or quantity of genomic DNA were suitable for molecular analysis for 51 cases of fibrous dysplasia. GNAS mutations were investigated by three techniques: high-resolution melting (exon 8), allele-specific PCR (exons 8 and 9) and/or direct DNA sequencing (exons 8 and 9). Fibrous dysplasia samples were classified blind to the GNAS mutation status into six histological subtypes as conventional, fibro-involutive, osteosclerosing, cementifying, osteocartilaginous and with prominent aneurysmal cystic changes. We also studied 14 cases of lowgrade osteosarcoma, 21 cases of ossifying fibroma, 3 cases of osteofibrous dysplasia, 1 case of osseous dysplasia of the jawbone and 1 post-traumatic lesion of the ribs. Twenty-three cases of fibrous dysplasia (45%) showed mutations of codon 201 (exon 8, p.R201H or p.R201C). No mutation was found on codon 227 (exon 9). GNAS mutations in conventional fibrous dysplasia were detected in the same proportion (47%) as in the other histological subtypes (47%, P ¼ 0.96), regardless of sex (P ¼ 0.44), age (P ¼ 0.90) and location (P ¼ 1). GNAS mutations were not detected in any other fibro-osseous lesions. The GNAS mutation was thus specific to fibrous dysplasia in the context of fibro-osseous lesions. The particular mosaicism of mutant and non-mutant cells within the lesion or the existence of other mutations not already described could explain the lack of GNAS mutation in cases of fibrous dysplasia. Investigating this mutation may constitute a valuable complementary diagnostic tool, despite its low sensitivity, particularly in unconventional morphologically different subtypes of fibrous dysplasia. Modern Pathology (2013) 26, 911-921;

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Section: Discussionmentioning

confidence: 99%

Diagnostic value of investigating GNAS mutations in fibro-osseous lesions: a retrospective study of 91 cases of fibrous dysplasia and 40 other fibro-osseous lesions

et al. 2013

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“…Nevertheless, the application of EDTA as a decalcifying agent in a routine setting is hampered by the long period required for incubation. 63 New methods such as decalcification in EDTA using a microwave oven and ultrasonic decalcification 64 are reported to considerably reduce the time of decalcification and preserve antigenic sites, DNA and mRNA. 63 In conclusion, overexpression and amplification of MDM2 and CDK4 are confirmed in low-grade osteosarcomas (parosteal osteosarcomas, low-grade central osteosarcomas and dedifferentiated lowgrade osteosarcomas).…”

Section: Discussionmentioning

confidence: 99%

“…63 New methods such as decalcification in EDTA using a microwave oven and ultrasonic decalcification 64 are reported to considerably reduce the time of decalcification and preserve antigenic sites, DNA and mRNA. 63 In conclusion, overexpression and amplification of MDM2 and CDK4 are confirmed in low-grade osteosarcomas (parosteal osteosarcomas, low-grade central osteosarcomas and dedifferentiated lowgrade osteosarcomas). The evaluation of MDM2 and CDK4 overexpression by immunohistochemistry appears to be a valuable diagnostic tool to distinguish low-grade osteosarcomas from lookalike benign fibrous and fibro-osseous lesions with quite good sensitivity.…”

Section: Discussionmentioning

confidence: 99%

MDM2 and CDK4 immunohistochemistry is a valuable tool in the differential diagnosis of low-grade osteosarcomas and other primary fibro-osseous lesions of the bone

Dujardin

Binh²,

Bouvier³

et al. 2011

Modern Pathology

203

122

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Low-grade osteosarcoma is a rare malignancy that may be subdivided into two main subgroups on the basis of location in relation to the bone cortex, that is, parosteal osteosarcoma and low-grade central osteosarcoma. Their histological appearance is quite similar and characterized by spindle cell stroma with low-to-moderate cellularity and well-differentiated anastomosing bone trabeculae. Low-grade osteosarcomas have a simple genetic profile with supernumerary ring chromosomes comprising amplification of chromosome 12q13-15, including the cyclindependent kinase 4 (CDK4) and murine double-minute type 2 (MDM2) gene region. Low-grade osteosarcoma can be confused with fibrous and fibro-osseous lesions such as fibromatosis and fibrous dysplasia on radiological and histological findings. We investigated MDM2-CDK4 immunohistochemical expression in a series of 72 low-grade osteosarcomas and 107 fibrous or fibro-osseous lesions of the bone or paraosseous soft tissue. The MDM2-CDK4 amplification status of low-grade osteosarcoma was also evaluated by comparative genomic hybridization array in 18 cases, and the MDM2 amplification status was evaluated by fluorescence in situ hybridization or quantitative real-time polymerase chain reaction in 31 cases of benign fibrous and fibro-osseous lesions. MDM2-CDK4 immunostaining and MDM2 amplification by fluorescence in situ hybridization or quantitative real-time polymerase chain reaction were investigated in a control group of 23 cases of primary high-grade bone sarcoma, including 20 conventional high-grade osteosarcomas, two pleomorphic spindle cell sarcomas/malignant fibrous histiocytomas and one leiomyosarcoma. The results showed that MDM2 and/or CDK4 immunoreactivity was present in 89% of lowgrade osteosarcoma specimens. All benign fibrous and fibro-osseous lesions and the tumors of the control group were negative for MDM2 and CDK4. These results were consistent with the MDM2 and CDK4 amplification results. In conclusion, immunohistochemical expression of MDM2 and CDK4 is specific and provides sensitive markers for the diagnosis of low-grade osteosarcomas, helping to differentiate them from benign fibrous and fibro-osseous lesions, particularly in cases with atypical radio-clinical presentation and/or limited biopsy samples.

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“…This, together with the difficulty in working with bony samples that contain a mixture of hard and soft tissues 2 has resulted in bone metastases being relatively understudied (compared with prostatic tissue obtained during therapeutic transurethral resection).…”

Section: Introductionmentioning

confidence: 99%

The comparative values of bone marrow aspirate and trephine for obtaining bone scan-targeted metastases from hormone-refractory prostate cancer

Brown¹,

Doğan

Ell

et al. 2002

Prostate Cancer Prostatic Dis

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Samples of metastatic prostate cancer to bone are difficult to obtain. The aim of this study was to compare the results of bone marrow aspirate and trephine biopsy for obtaining metastatic hormone-refractory prostate cancer (HRPC) samples using previous diagnostic planar 99 m Tc-HDP bone scans to guide the procedure. All samples taken were for the purposes of research and molecular studies on HRPC. Twenty patients with HRPC had bone marrow aspirate and trephines taken from lesions in the posterior superior iliac spine or sacro-iliac region when shown on diagnostic 99 m Tc-HDP bone scans. Three patients also underwent plain X-ray, 18F-positron emission tomography bone scan, pelvic MRI scan and 99 m Tc nanocolloid bone marrow scans. These images were used to assess if the extra imaging information provided, such as three-dimensional localisation of the bone metastases, was of value for target bone metastases. Cancer cells were obtained in 15/20 (75%) cases in which a trephine biopsy was attempted and 0/20 of cases in which a bone marrow aspiration was attempted. The additional information provided by the range of other imaging investigations was of little benefit in obtaining tumour samples, but did suggest why negative biopsies were obtained in some cases after targeting with planar bone scans. We recommend the use of bone marrow trephine biopsy alone, guided by previous diagnostic 99 m Tc planar bone scan as a practical method to obtain prostate cancer cells from bone metastases. Keywords: 99 m Tc-bone scan; 99 m Tc-nanocolloid bone marrow scan; bone marrow aspiration; bone marrow trephine; Fluorine18 positron emission tomography bone scan; hormone-resistant prostate cancer; pelvic magnetic resonance imaging scan

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Effect of Bone Decalcification Procedures on DNA In Situ Hybridization and Comparative Genomic Hybridization: EDTA Is Highly Preferable to a Routinely Used Acid Decalcifier

Cited by 130 publications

References 16 publications

Diagnostic value of investigating GNAS mutations in fibro-osseous lesions: a retrospective study of 91 cases of fibrous dysplasia and 40 other fibro-osseous lesions

Diagnostic value of investigating GNAS mutations in fibro-osseous lesions: a retrospective study of 91 cases of fibrous dysplasia and 40 other fibro-osseous lesions

MDM2 and CDK4 immunohistochemistry is a valuable tool in the differential diagnosis of low-grade osteosarcomas and other primary fibro-osseous lesions of the bone

The comparative values of bone marrow aspirate and trephine for obtaining bone scan-targeted metastases from hormone-refractory prostate cancer

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