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1.
J Plast Reconstr Aesthet Surg ; 62(7): 889-95, 2009 Jul.
Article in English | MEDLINE | ID: mdl-18440289

ABSTRACT

The local treatment protocol of preoperative radiotherapy in head and neck cancer treatment at the Karolinska University Hospital has resulted in a unique cohort of preoperatively high dose-irradiated patients. In total 216 consecutive patients were reviewed, of whom 221 free flaps, for head and neck cancer reconstruction, were operated between 1984 and 2002. In 194 cases radiotherapy was administered preoperatively and 27 operations were performed without prior radiation. The radiation dose was 64 Gy in 147 cases, 54 Gy or less in 45 cases and uncertain in two cases. In order to study whether the time elapsed between the end of radiotherapy and surgery had any significance regarding postoperative events, the cohort was subsequently divided into three groups: patients operated on within 4 weeks (n=27), between 4 and 6 weeks (n=88) and more than 6 weeks (n=78) after the last radiotherapy session. Postoperative complications were analysed in relation to preoperative dose and timing of radiotherapy. Preoperative radiotherapy was related to an increased risk of free flap necrosis as 22 complete and eight partial flap necroses occurred in the group that had received preoperative radiotherapy and none were observed in the non-irradiated group (P<0.05). Furthermore, a linear trend of increased flap loss (P<0.001), infections (P<0.001) and delayed wound healing (P<0.001) was seen when time increased between the last radiotherapy session and surgery. The largest increase in all complication rates was seen when more than 6 weeks elapsed between last radiotherapy session and surgery. Postoperative complications were independent of the radiation dose given. Our data show an increased morbidity in free flap surgery in the head and neck region after preoperative radiotherapy. Furthermore, time elapsed between the last radiotherapy session and surgery is associated with the risk of developing postoperative complications. We strongly suggest that free flap reconstruction should be performed within 6 weeks of the last radiotherapy session.


Subject(s)
Head and Neck Neoplasms/radiotherapy , Postoperative Complications/etiology , Surgical Flaps , Aged , Female , Head and Neck Neoplasms/surgery , Humans , Male , Middle Aged , Necrosis/etiology , Postoperative Complications/pathology , Radiotherapy Dosage , Plastic Surgery Procedures , Retrospective Studies , Surgical Flaps/pathology , Time Factors , Treatment Outcome
2.
Kidney Int ; 60(5): 1666-75, 2001 Nov.
Article in English | MEDLINE | ID: mdl-11703583

ABSTRACT

BACKGROUND: Secondary hyperparathyroidism (HPT) is characterized by inappropriate control of parathyroid hormone (PTH) secretion and asymmetric hyperplasia of the parathyroid glands. Receptors for calcium and vitamin D are involved in the control of secretion, as well as parathyroid cell proliferation. Defective receptor mechanisms therefore may play a role in the pathogenensis of secondary HPT. Previous studies have shown that the expression of calcium receptor (CaR), calcium-sensing receptor (CAS) and vitamin D receptor (VDR) protein, and mRNA is decreased in hyperplastic parathyroid glands of secondary HPT when compared with normal parathyroid glands. METHODS: Thirty-six hyperplastic glands from 18 patients with secondary hyperparathyroidism were analyzed with in situ hybridization in order to investigate the expression of CaR, CAS, VDR, and PTH mRNAs in the same specimens. In nine nodular parathyroid glands, it was possible to make a comparison between the expression of these mRNAs in nodular and internodular areas. RESULTS: The level of CaR was in the same order of magnitude in the hyperplastic glands and in the biopsies of normal parathyroid, whereas the levels of CAS, VDR and PTH were clearly reduced in the hyperplastic glands. There was a positive correlation between the expression of CaR and CAS (P = 0.02). Otherwise, no correlations between CaR, CAS, VDR, and PTH mRNAs were found. The expression of all four genes was highly variable as well between different glands as within individual glands. CONCLUSION: The expression of mRNAs for receptors of importance in the control of PTH secretion and parathyroid cell proliferation is heterogeneously decreased in parathyroid glands of secondary HPT. The expression pattern corroborates earlier studies in which it has been assumed that each nodule in secondary HPT is of monoclonal origin, but that the monoclonal origin of each nodule is independent.


Subject(s)
Calcium-Binding Proteins/genetics , Hyperparathyroidism, Secondary/metabolism , Parathyroid Glands/metabolism , RNA, Messenger/analysis , Receptors, Calcitriol/genetics , Receptors, Cell Surface/genetics , Adult , Aged , Child , Female , Glyceraldehyde-3-Phosphate Dehydrogenases/genetics , Humans , Male , Middle Aged , Parathyroid Hormone/genetics , Receptors, Calcium-Sensing
3.
Eur J Endocrinol ; 145(4): 415-20, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11580998

ABSTRACT

BACKGROUND: Most patients who have been surgically treated for secondary hyperparathyroidism (HPT) harbor at least one pathological parathyroid gland with a tumor of monoclonal origin. OBJECTIVE: To elucidate the underlying genetic mechanisms behind secondary HPT, by studying a panel of such tumors for numerical alterations. METHODS: Sixteen parathyroid glands from eight patients (median age 58 years, range 31-74 years), were screened for numerical chromosomal imbalances, using comparative genomic hybridization (CGH). Mutation analysis of the multiple endocrine neoplasia type 1 gene (MEN1) was also performed by sequencing of the coding region. RESULTS: The results show that gross chromosomal alterations occur rarely in secondary HPT. In one of the three glands analyzed from one patient, a complete loss of chromosome 11 was detected. This gland also had an inactivating nonsense mutation, E469X, of the MEN1 gene. The mutation was present neither in the other two glands, nor in the constitutional tissue of the same patient, thus confirming its somatic origin. CONCLUSIONS: The relative lack of numerical chromosomal alterations would suggest that more discrete genetic alterations are responsible for the monoclonal growth in the majority of cases of secondary HPT. Furthermore, somatic inactivation of the MEN1 tumor suppressor gene contributes to the tumorigenesis in a small proportion of the cases.


Subject(s)
Gene Silencing , Homozygote , Hyperparathyroidism, Secondary/genetics , Neoplasm Proteins/genetics , Proto-Oncogene Proteins , Adult , Aged , Base Sequence/genetics , Chromosomes, Human, Pair 11 , Codon, Nonsense , Female , Gene Deletion , Humans , Male , Middle Aged , Nucleic Acid Hybridization
4.
Eur J Haematol ; 66(5): 297-304, 2001 May.
Article in English | MEDLINE | ID: mdl-11422408

ABSTRACT

Spectral karyotyping (SKY) on metaphase spreads from 15 high hyperdiploid (>51 chromosomes) childhood acute lymphoblastic leukemias (ALL), which typically display a poor chromosome morphology, was performed in order to investigate the pattern of numerical abnormalities, reveal the chromosomal origin of marker chromosomes, and identify translocations and other interchromosomal rearrangements not detected by G-banding analysis. In all cases the numerical changes could be fully characterized, and a non-random pattern of chromosomal gain was identified, with chromosomes X, 21, 14, 17, 6, 18, 4, and 10 being most frequently gained. The numerical changes had been partly misinterpreted in 12 of the 15 ALL patients using G-banding, and the present study hence emphasizes the importance of SKY in identifying such anomalies, some of which, i.e. +4 and +10, have been suggested to be prognostically important. The chromosomal origin of all marker chromosomes and of seven structural rearrangements, one of which was the prognostically important Philadelphia chromosome, could be identified. Five rearrangements [der(1)t(1;14)(q32;q21), der(2)t(2;8)(q36;?), der(3)t(2;3)(q21;?), der(8)t(8;14)(?;?), and t(9;21)(q12;q22)] have previously not been reported in ALL, emphasizing the value of SKY in identifying novel chromosomal rearrangements.


Subject(s)
Chromosome Aberrations , Diploidy , Karyotyping/methods , Precursor Cell Lymphoblastic Leukemia-Lymphoma/genetics , Child , Child, Preschool , Chromosome Banding , Chromosomes, Human, Pair 10 , Chromosomes, Human, Pair 14 , Chromosomes, Human, Pair 17 , Chromosomes, Human, Pair 18 , Chromosomes, Human, Pair 21 , Chromosomes, Human, Pair 4 , Chromosomes, Human, Pair 6 , Female , Humans , In Situ Hybridization, Fluorescence , Infant , Male , X Chromosome
5.
Cancer Genet Cytogenet ; 126(1): 1-7, 2001 Apr 01.
Article in English | MEDLINE | ID: mdl-11343771

ABSTRACT

Comparative genomic hybridization (CGH) studies have shown that chromosome 8 is a frequent target for chromosomal aberrations in breast cancer. We characterized these aberrations of chromosome 8 in 16 breast cancer cell lines (BT-474, BT-549, CAMA-1, DU-4475, MCF-7, MDA-MB-134, MDA-MB-157, MDA-MB-361, MDA-MB-415, MDA-MB-436, MPE600, SK-BR-3, T-47D, UACC-812, UACC-893 and ZR-75-1) by CGH, fluorescence in situ hybridization (FISH) with arm- and locus-specific probes, and spectral karyotyping (SKY). Chromosome 8 was structurally abnormal in 13 of 16 cell lines. Loss of 8p was detected in nine cell lines, gain of entire 8q in six cell lines, 8q21-qter in three, 8q23-qter in two, and 8q12-qter and 8p21-q21 in one cell line. Extra copies of the C-MYC oncogene were found in 11 cell lines, but high-level amplification only in SK-BR-3. Derivative chromosomes including material from chromosomes 8 were complex, and the breakpoints were strikingly dissimilar. Chromosome 11 was the most frequent translocation partner with chromosome 8 (in 7 cell lines). Isochromosomes and/or isoderivative 8q were found in four cell lines. The high frequency and complexity of alterations at 8q indicate a significant pathogenetic role in breast cancer. The high-level amplification of c-myc is less common than previously thought.


Subject(s)
Breast Neoplasms/genetics , Chromosome Aberrations , Chromosomes, Human, Pair 8 , Breast Neoplasms/pathology , Female , Humans , Karyotyping/methods , Nucleic Acid Hybridization/methods , Tumor Cells, Cultured
6.
Proc Natl Acad Sci U S A ; 98(8): 4605-10, 2001 Apr 10.
Article in English | MEDLINE | ID: mdl-11274374

ABSTRACT

Although the systemic administration of a number of different gene products has been shown to result in the inhibition of angiogenesis and tumor growth in different animal tumor models, the relative potency of those gene products has not been studied rigorously. To address this issue, recombinant adenoviruses encoding angiostatin, endostatin, and the ligand-binding ectodomains of the vascular endothelial growth factor receptors Flk1, Flt1, and neuropilin were generated and used to systemically deliver the different gene products in several different preexisting murine tumor models. Single i.v. injections of viruses encoding soluble forms of Flk1 or Flt1 resulted in approximately 80% inhibition of preexisting tumor growth in murine models involving both murine (Lewis lung carcinoma, T241 fibrosarcoma) and human (BxPC3 pancreatic carcinoma) tumors. In contrast, adenoviruses encoding angiostatin, endostatin, or neuropilin were significantly less effective. A strong correlation was observed between the effects of the different viruses on tumor growth and the activity of the viruses in the inhibition of corneal micropocket angiogenesis. These data underscore the need for comparative analyses of different therapeutic approaches that target tumor angiogenesis and provide a rationale for the selection of specific antiangiogenic gene products as lead candidates for use in gene therapy approaches aimed at the treatment of malignant and ocular disorders.


Subject(s)
Neovascularization, Pathologic , Proto-Oncogene Proteins/physiology , Receptor Protein-Tyrosine Kinases/physiology , Receptors, Growth Factor/physiology , Transfection , Adenoviridae/genetics , Amino Acid Sequence , Animals , Cell Division/genetics , Evaluation Studies as Topic , Humans , Mice , Molecular Sequence Data , Neoplasms/blood supply , Proto-Oncogene Proteins/genetics , Receptor Protein-Tyrosine Kinases/genetics , Receptors, Growth Factor/genetics , Receptors, Vascular Endothelial Growth Factor , Tumor Cells, Cultured , Vascular Endothelial Growth Factor Receptor-1
7.
Int J Mol Med ; 6(4): 469-74, 2000 Oct.
Article in English | MEDLINE | ID: mdl-10998441

ABSTRACT

Pheochromocytoma and its extra-adrenal counterpart paraganglioma are rare catecholamine producing tumors which usually occur sporadically but may also be a part of neuroendocrine tumor syndromes such as multiple endocrine neoplasia type 2A (MEN 2A). Activating mutations of the RET proto-oncogene which is the underlying cause of MEN 2A, is also seen in approximately 10% of sporadic pheochromocytomas. Glial cell line derived neurotrophic factor (GDNF) and neurturin (NTN) have been shown to function as independent ligands to RET, binding in a complex with the membrane-bound receptors GFRalpha-1 and GFRalpha-2 respectively. Here we have investigated the mRNA expression of RET and its ligand complexes, GDNF/GFRalpha-1 and NTN/GFRalpha-2, in a panel of pheochromocytomas and paragangliomas using mRNA in situ hybridization. RET expression was evident in normal adrenal medulla, and in 13/15 pheochromocytomas, including 5/5 MEN 2A associated tumors, but only in 1/10 paragangliomas. The frequent expression of RET in the pheochromocytomas suggest that this gene might be involved in the tumorigenesis. However, no expression of GDNF/GFRalpha-1 or NTN/GFRalpha-2 could be detected in any of the 25 tumors analyzed, suggesting that these ligand complexes are not important in the development of pheochromocytoma or paraganglioma.


Subject(s)
Drosophila Proteins , Genes/genetics , Paraganglioma/genetics , Pheochromocytoma/genetics , Adolescent , Adult , Aged , Female , Gene Expression Regulation, Neoplastic , Glial Cell Line-Derived Neurotrophic Factor , Glial Cell Line-Derived Neurotrophic Factor Receptors , Humans , In Situ Hybridization , Male , Middle Aged , Nerve Growth Factors/genetics , Nerve Tissue Proteins/genetics , Neurturin , Paraganglioma/pathology , Pheochromocytoma/pathology , Proto-Oncogene Mas , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins c-ret , RNA, Messenger/genetics , RNA, Messenger/metabolism , Receptor Protein-Tyrosine Kinases/genetics
8.
Clin Endocrinol (Oxf) ; 53(1): 85-92, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10931084

ABSTRACT

OBJECTIVE: Hyperparathyroidism (HPT) is a common endocrine disorder. Several loci of genetic interest have been identified in parathyroid tumours, including the MEN1 gene locus at 11q13; the HPT-JT region at 1q21-q32; and a putative tumour suppressor gene on 1p. We analysed these intervals, which harbour known genes or putative loci associated with familial hyperparathyroidism, in order to clarify the involvement of the respective regions in parathyroid tumourigenesis. DESIGN: We performed loss of heterozygosity (LOH) studies on 33 sporadic parathyroid tumours using a PCR based technique. A total of 22 microsatellite markers were used to analyse loci at 11q13, 1q21-q32 and 1p. Ten markers located distal on 1p, eight markers encompassed the HPT-JT region at 1q21-q32 and four markers surrounded the MEN1 gene locus at 11q13. MEN1 mutations were screened for using Single Strand Conformation Polymorphism analysis (SSCP) and automated sequencing of SSCP variants. PATIENTS: Thirty-three parathyroid glands and the corresponding blood samples were obtained from 33 patients (26 females and seven males) who underwent parathyroidectomy for primary hyperparathyroidism. RESULTS: Loss of heterozygosity was detected in 13 of 33 (39%) cases at 11q13, 6 of 33 (18%) cases at 1p, and in three of 33 (9%) cases at 1q (in conjunction with 1p loss). Only one of the 18 tumours in which LOH was detected, showed LOH at both chromosome 1 and chromosome 11. Additionally, those tumours found to exhibit LOH at 11q13 were screened for MEN1 mutations using single strand conformation polymorphism analysis (SSCP) and automated sequencing. Nine novel somatic mutations were found on the remaining allele in 13 (69%) tumours. CONCLUSIONS: This study consolidates the role of multiple loci in the pathogenesis of sporadic parathyroid tumours. The results indicate that there are at least two genetic loci involved in sporadic parathyroid tumourigenesis on chromosome 1, one of which has been linked to the distinct familial parathyroid condition, hyperparathyroidism-jaw tumour (HPT-JT) syndrome. The high frequency of loss of heterozygosity at 1p suggests the presence of a tumour suppressor at this locus.


Subject(s)
Adenoma/genetics , Chromosomes, Human, Pair 11/genetics , Chromosomes, Human, Pair 1/genetics , Loss of Heterozygosity , Neoplasm Proteins/genetics , Parathyroid Neoplasms/genetics , Proto-Oncogene Proteins , Female , Humans , Male , Multiple Endocrine Neoplasia Type 1/genetics , Mutation , Polymorphism, Single-Stranded Conformational
9.
Am J Pathol ; 157(2): 579-86, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10934160

ABSTRACT

In this study we have characterized chromosomal imbalances in a panel of 29 parathyroid carcinomas using comparative genomic hybridization (CGH). The most frequently detected imbalances were losses of 1p and 13q that were seen in >40% of the cases. The commonly occurring regions of loss were assigned to 1p21-p22 (41%), 13q14-q31 (41%), 9p21-pter (28%), 6q22-q24 (24%), and 4q24 (21%), whereas gains preferentially involved 19p (45%), Xc-q13 (28%), 9q33-qter (24%), 1q31-q32 (21%) and 16p (21%). The distribution of CGH alterations supports the idea of a progression of genetic events in the development of parathyroid carcinoma, where gains of Xq and 1q would represent relatively early events that are followed by loss of 13q, 9p, and 1p, and by gain of 19p. A sex-dependent distribution was also evident for two of the common alterations with preferential gain of 1q in female cases and of Xq in male cases. When the CGH profiles for the 29 carcinomas were compared with our previously published results for sporadic parathyroid adenomas, highly significant differences were revealed. Loss of 1p, 4q, and 13q as well as gains of 1q, 9q, 16p, 19p and Xq were significantly more common in the carcinomas than in the adenomas. In contrast, loss of the 11q13 region, which is the most common CGH abnormality in sporadic adenomas, was not detected in any of the carcinomas. Taken together, the findings identify several candidate locations for tumor suppressor genes and oncogenes that are potentially involved in parathyroid carcinogenesis.


Subject(s)
Chromosome Aberrations , Parathyroid Neoplasms/genetics , Adenoma/genetics , Adenoma/pathology , Adult , Aged , Female , Humans , Male , Middle Aged , Nucleic Acid Hybridization/methods , Parathyroid Glands/metabolism , Parathyroid Glands/pathology , Parathyroid Neoplasms/pathology
10.
Genes Chromosomes Cancer ; 28(3): 308-17, 2000 Jul.
Article in English | MEDLINE | ID: mdl-10862037

ABSTRACT

Breast cancer cell lines have been widely used as models in functional and therapeutical studies, but their chromosomal alterations are not well known. We characterized the chromosomal aberrations in 15 commonly used human breast carcinoma cell lines (BT-474, BT-549, CAMA-1, DU4475, MCF7, MDA-MB-134, MDA-MB-157, MDA-MB-361, MDA-MB-436, MPE600, SK-BR-3, T-47D, UACC-812, UACC-893, and ZR-75-1) by comparative genomic hybridization (CGH) and spectral karyotyping (SKY). By CGH the most frequent gains were detected at 1q, 8q, 20q, 7, 11q13, 17q, 9q, and 16p, whereas losses were most common at 8p, 11q14-qter, 18q, and Xq. SKY revealed a multitude of structural and numerical chromosomal aberrations. Simple translocations, typically consisting of entire translocated chromosome arms, were the most common structural aberrations. Complex marker chromosomes included material from up to seven different chromosomes. Evidence for a cytogenetic aberration not previously described in breast cancer, the isoderivative chromosome, was found in two cell lines. Translocations t(8;11), t(12;16), t(1;16), and t(15;17) were frequently found, although the resulting derivative chromosomes and their breakpoints were strikingly dissimilar. The chromosomes most frequently involved in translocations were 8, 1, 17, 16, and 20. An excellent correlation was found between the number of translocation events found by SKY in the individual cell lines, and the copy number gains and losses detected by CGH, indicating that the majority of translocations are unbalanced. Genes Chromosomes Cancer 28:308-317, 2000.


Subject(s)
Breast Neoplasms/genetics , Chromosome Aberrations/genetics , DNA, Neoplasm/genetics , Chromosome Disorders , Genes, myc/genetics , Humans , In Situ Hybridization, Fluorescence , Karyotyping/methods , Nucleic Acid Hybridization/genetics , Nucleic Acid Hybridization/methods , Tumor Cells, Cultured
11.
Eur J Endocrinol ; 142(6): 643-9, 2000 Jun.
Article in English | MEDLINE | ID: mdl-10822229

ABSTRACT

OBJECTIVE: Mutations in the RET proto-oncogene are found in about one third of sporadic medullary thyroid carcinomas (MTCs), mostly affecting codon 918. Glial cell line derived neurotropic factor (GDNF) and its membrane-bound GDNF family receptor alpha (GFRalpha-1), as well as neurturin (NTN) and its membrane-bound receptor GFRalpha-2 form a complex with the RET product, a receptor tyrosine kinase, resulting in downstream signaling to the nucleus. DESIGN: To elucidate the role of these RET ligands in MTC tumorigenesis, their expression was determined in 15 MTC samples, one papillary thyroid carcinoma (PTC) and three normal thyroid tissue specimens. METHODS: The mRNA expression of RET, GDNF, GFRalpha-1, NTN and GFRalpha-2 was investigated by mRNA in situ hybridization, and confirmed by reverse transcription-PCR analysis. RESULTS: None of the five genes was expressed in the normal thyroids or in the PTC. All MTCs showed expression of RET, 13 expressed GDNF, 12 expressed GFRalpha-1 and 9 expressed NTN and GFRalpha-2. In 7 of the tumors RET, GDNF and GFRalpha-1 were expressed at high levels, and in five of these seven tumors NTN and GFRalpha-2 genes were also expressed at high levels. The high level of expression was preferentially seen in tumor cells adjacent to stroma and connective tissue. All MTCs without expression of the RET ligands harbored the RET codon 918 mutation. CONCLUSIONS: The results suggest that this signaling pathway is important for MTC development, and that it may be activated by expression of the RET ligand complexes by the tumor cells themselves.


Subject(s)
Carcinoma, Medullary/metabolism , Drosophila Proteins , Nerve Growth Factors/metabolism , Nerve Tissue Proteins/metabolism , Proto-Oncogene Proteins/metabolism , Receptor Protein-Tyrosine Kinases/metabolism , Thyroid Neoplasms/metabolism , Adult , Aged , Codon/genetics , Female , Glial Cell Line-Derived Neurotrophic Factor , Glial Cell Line-Derived Neurotrophic Factor Receptors , Humans , In Situ Hybridization , Ligands , Male , Middle Aged , Mutation , Nerve Growth Factors/genetics , Nerve Tissue Proteins/genetics , Neurturin , Proto-Oncogene Mas , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins c-ret , RNA, Messenger/metabolism , Receptor Protein-Tyrosine Kinases/genetics , Reverse Transcriptase Polymerase Chain Reaction
13.
Hematol J ; 1(2): 95-101, 2000.
Article in English | MEDLINE | ID: mdl-11920176

ABSTRACT

INTRODUCTION: Plasma cell leukaemia is a rare disorder that usually carries an aggressive course with a rapidly fatal outcome. A variety of chromosomal abnormalities have been reported in plasma cell leukaemia but the clinical significance of an abnormal karyotype is still unclear. MATERIALS AND METHODS: We have applied the molecular cytogenetic techniques multicolour spectral karyotyping and microdissection in combination with fluorescence in situ hybridization on metaphases from a patient with primary plasma cell leukaemia and a fatal outcome. RESULTS AND CONCLUSION: The chromosome analysis showed severe hypodiploidy and 12 marker chromosomes. Identification of the structural rearrangements was not possible using routine cytogenetic methods. Utilizing the methods above, all marker chromosomes could be identified in detail and the karyotype was shown to be very complex. Forty-three breakpoints were found, and 25 could be identified at the band level, among others 14q32 where the immunoglobulin heavy chain locus is situated. Thus, these techniques provide the opportunity to resolve very complex chromosomal changes in a way that has not been previously possible and will consequently be of great importance in the search for hot spots that may harbour new cancer genes.


Subject(s)
Chromosome Aberrations , Leukemia, Plasma Cell/genetics , Adult , Bone Marrow Cells/pathology , Chromosome Banding , Chromosome Mapping , Fatal Outcome , Humans , In Situ Hybridization, Fluorescence , Karyotyping , Leukemia, Plasma Cell/pathology , Male
14.
Surgery ; 126(6): 1183-7, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10598205

ABSTRACT

BACKGROUND: The incidence of parathyroid cancer in patients with hyperparathyroidism is less than 1%. However, these few cases cause diagnostic problems in the absence of clear-cut invasion of adjacent organs or metastasis. New markers are needed to increase diagnostic accuracy. METHODS: Thirty-one parathyroid tumors from patients with primary hyperparathyroidism were collected worldwide. Eighteen tumors were classified as unequivocal cancers, whereas 13 tumors were considered equivocal because of a lack of infiltrative growth or evidence of recurrence. Paraffin sections were hybridized with a 35S-labeled riboprobe complementary to gelatinase A mRNA, dipped in photographic emulsion, developed, counterstained, and then evaluated by light- and dark-field microscopy. RESULTS: Fourteen of the 18 unequivocal parathyroid cancers expressed gelatinase A, as compared with the equivocal tumors, of which only 4 of 13 showed expression. The strongest hybridization signal was seen in stromal cells at the tumor border, most likely fibroblasts and macrophages. No expression was detected in tumor cells. CONCLUSIONS: Invasive growth of many tumors is facilitated by proteolytic enzymes, such as gelatinase A. The presence of gelatinase A mRNA in parathyroid tumors strengthens the suspicion of malignancy but cannot be used as a definitive marker of malignancy.


Subject(s)
Carcinoma/enzymology , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Neoplastic , Matrix Metalloproteinase 2/genetics , Parathyroid Neoplasms/enzymology , Adult , Aged , Aged, 80 and over , Carcinoma/genetics , Carcinoma/pathology , DNA, Complementary , Fibroblasts/enzymology , Humans , In Situ Hybridization , Middle Aged , Parathyroid Neoplasms/genetics , Parathyroid Neoplasms/pathology , RNA, Messenger/analysis
15.
J Clin Endocrinol Metab ; 84(10): 3775-80, 1999 Oct.
Article in English | MEDLINE | ID: mdl-10523029

ABSTRACT

In this study 44 parathyroid tumors from 26 sporadic cases, 10 cases previously given irradiation to the neck, and 8 familial cases were screened for sequence copy number alterations by comparative genomic hybridization. In the sporadic adenomas, commonly occurring minimal regions of loss could be defined to chromosome 11 (38%), 15q15-qter (27%), and 1p34-pter (19%), whereas gains preferentially involved 19p13.2-pter (15%) and 7pter-qter (12%). Multiple aberrations were found in sporadic tumors with a somatic mutation and/or loss of heterozygosity of the MEN1 gene. The irradiation-associated tumors also showed multiple comparative genomic hybridization alterations and frequent losses of 11q (50%), and subsequent analysis of the MEN1 gene demonstrated mutations in 4 of 8 cases (50%). The adenomas from familial cases showed few alterations, and in 3 of these tumors a gain of 19p13.2-pter was seen as the only aberration. In this study numerical copy number alterations were frequently detected in sporadic and irradiation-associated parathyroid adenomas, although these tumors are benign. The majority of these alterations were found in tumors with confirmed involvement of the MEN1 gene locus in agreement with a role of the MEN1 gene in genomic stability. Furthermore, the frequent occurrence of MEN1 mutations (50%) in irradiation-associated parathyroid tumors suggests that inactivation of the MEN1 gene is an important genetic alteration involved in the development of parathyroid tumors in postirradiation patients.


Subject(s)
Parathyroid Neoplasms/genetics , Adenoma/genetics , Adult , Aged , Chromosome Aberrations , Chromosome Mapping , DNA Mutational Analysis , Female , Humans , Loss of Heterozygosity , Male , Middle Aged , Multiple Endocrine Neoplasia Type 1/genetics , Mutation/genetics , Neck/radiation effects , Neoplasms, Radiation-Induced/genetics , Nucleic Acid Hybridization
16.
Clin Endocrinol (Oxf) ; 50(2): 191-6, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10396361

ABSTRACT

BACKGROUND AND OBJECTIVES: Familial hyperparathyroidism may occur as familial isolated hyperparathyroidism (FIHP) or as part of an inherited syndrome, in particular multiple endocrine neoplasia types 1 and 2A (MEN1, MEN2A) and hyperparathyroidism-jaw tumour (HPT-JT) syndrome. The localization of the genes responsible for these syndromes has enabled genetic screening of families with primary hyperparathyroidism (PHPT) to be carried out. This has important clinical implications in terms of individual follow-up and management. We previously reported a large FIHP family with an increased risk of parathyroid cancer and excluded its linkage to MEN1, MEN2 and PTH genes. Here we re-analysed this family and performed genetic linkage to the HPT-JT locus in chromosome 1q21-q32. Loss of heterozygosity studies of 1q21-q32, 11q13 and X chromosome were also performed. PATIENTS AND DESIGN: We studied 19 family members, aged 6-63 years. High molecular weight DNA was isolated from peripheral blood samples from 17 family members. For the two deceased individuals, DNA was extracted from normal paraffin embedded tissues. MEASUREMENTS: All individuals (except two deceased patients) had serum corrected calcium, inorganic phosphate, intact PTH, prolactin and various pancreatic hormones, measured on fasting blood samples. Twenty microsatellite markers were examined for the 1q21-q32 region, the locus for the HPT-JT gene. Genetic polymorphisms were determined by polymerase chain reaction amplification of genomic DNA and genetic linkage analysis was performed. Loss of heterozygosity studies were performed using paraffin-embedded parathyroid tissues from four affected members. RESULTS: Seven of the eight affected family members included in this study had biochemical evidence of PHPT and surgically proven parathyroid tumours. Indication of linkage of the disease to the HPT-JT locus was demonstrated with a maximum lod score of 2.32 by two-points linkage analysis. Linkage data were supported by multi-point analysis which gave a maximum lod score of 2.7. Meiotic recombinations detected in one affected individual narrowed the region to 26 cM. As a result of the genetic findings, we re-screened the living family members by orthopantomograph and renal ultrasound, and identified two jaw lesions in two gene carriers. One affected family member demonstrated polycystic kidney disease, thus establishing the association between the two conditions. A reduced penetrance of HPT in females was evident, in agreement with our previous study. No allelic deletion was detected in any tumour at 1q21-q32, 11q13 or X chromosome. CONCLUSIONS: This study illustrates the usefulness and importance of genetic studies in familial isolated hyperparathyroidism families. Our clinical and genetic findings indicate that this previously reported familial isolated hyperparathyroidism family has hyperparathyroidism-jaw tumour syndrome.


Subject(s)
Chromosomes, Human, Pair 1 , Genetic Linkage , Hyperparathyroidism/genetics , Jaw Neoplasms/genetics , Adolescent , Adult , Child , Female , Humans , Jaw Neoplasms/diagnosis , Lod Score , Loss of Heterozygosity , Male , Middle Aged , Pedigree , Syndrome
17.
Biochem Biophys Res Commun ; 257(3): 891-4, 1999 Apr 21.
Article in English | MEDLINE | ID: mdl-10208880

ABSTRACT

Endothelial growth factors have become the target of intense research since the initial discovery of vascular endothelial growth factor (VEGF/VPF). At present, VEGF is established as a major inducer of angiogenesis in normal and pathological conditions. Recently several new members in the VEGF family have been described; VEGF-B/VRF, VEGF-C and VEGF-D. VEGF-D is most closely related to VEGF-C by virtue of the presence of N- and C-terminal extensions that are not found in other VEGF family members. We have here examined the expression pattern of vegf-d mRNA with in situ hybridization in developing and adult mice. This shows a restricted expression pattern, with high levels mainly in lung tissue. The expression in embryonic lung is upregulated prior to birth. Expression of vegf-d in other tissues, as well as in lung tissue of the E14 embryo, was either low or absent. This suggests that VEGF-D may be of special relevance for the vascularization of lung tissue during the last trimester of fetal development.


Subject(s)
Endothelial Growth Factors/genetics , Gene Expression Regulation, Developmental , Lung/embryology , Up-Regulation , Animals , Embryo, Mammalian/metabolism , Embryonic and Fetal Development , Female , In Situ Hybridization , Kidney/metabolism , Lung/growth & development , Lung/metabolism , Male , Mice , Mice, Inbred Strains , Oligonucleotides/genetics , Organ Specificity , RNA, Messenger/metabolism , Seminiferous Tubules/metabolism , Spine/embryology , Spine/metabolism , Thymus Gland/metabolism , Vascular Endothelial Growth Factor D
18.
World J Surg ; 23(1): 68-74, 1999 Jan.
Article in English | MEDLINE | ID: mdl-9841766

ABSTRACT

Assessment of the malignant potential of parathyroid tumors in the absence of metastases can be difficult using morphologic criteria alone. In this study we have examined a total of 58 parathyroid tumors (31 benign, 15 malignant, and 12 equivocal) from 54 patients using immunohistochemistry with monoclonal antibodies directed against the retinoblastoma (RB) protein and the cell cycle-associated antigen Ki-67 to evaluate their role as diagnostic markers. RB protein immunoreactivity was not useful for distinguishing between benign and malignant parathyroid tumors. Analysis of the proliferation marker Ki-67 showed that there was a trend toward more intense staining in the malignant cases. The Ki-67 labeling index was highest in the parathyroid cancers (median 33) and lowest in the sporadic primary adenomas (median 2). An observation that might have clinical implications is that tumors from patients with familial hyperparathyroidism linked to chromosome 1q showed a high Ki-67 index, indicating strong proliferative activity (median 25). This correlates well with the clinical observation of tumors with malignant potential in this syndrome. Because of the considerable overlap between groups of tumors, Ki-67 is not suitable for definitive differentiation between benign and malignant tumors. However, Ki-67 may give valuable information about which patients should be followed more closely.;1999>


Subject(s)
Biomarkers, Tumor/analysis , Ki-67 Antigen/analysis , Parathyroid Neoplasms/pathology , Retinoblastoma Protein/analysis , Antibodies, Monoclonal , Humans , Hyperparathyroidism, Secondary/genetics , Immunohistochemistry , Parathyroid Neoplasms/complications , Parathyroid Neoplasms/immunology , Statistics, Nonparametric
19.
Cytogenet Cell Genet ; 87(3-4): 225-32, 1999.
Article in English | MEDLINE | ID: mdl-10702678

ABSTRACT

Human prostate cancer is characterized by multiple gross chromosome alterations involving several chromosome regions. However, the specific genes involved in the development of prostate tumors are still largely unknown. Here we have studied the chromosome composition of the three established prostate cancer cell lines, LNCaP, PC-3, and DU145, by spectral karyotyping (SKY). SKY analysis showed complex karyotypes for all three cell lines, with 87, 58/113, and 62 chromosomes, respectively. All cell lines were shown to carry structural alterations of chromosomes 1, 2, 4, 6, 10, 15, and 16; however, no recurrent breakpoints were detected. Compared to previously published findings on these cell lines using comparative genomic hybridization, SKY revealed several balanced translocations and pinpointed rearrangement breakpoints. The SKY analysis was validated by fluorescence in situ hybridization using chromosome-specific, as well as locus-specific, probes. Identification of chromosome alterations in these cell lines by SKY may prove to be helpful in attempts to clone the genes involved in prostate cancer tumorigenesis.


Subject(s)
Chromosome Aberrations/genetics , Karyotyping , Prostatic Neoplasms/genetics , Aneuploidy , Chromosome Breakage/genetics , Chromosomes, Human, Pair 8/genetics , Humans , In Situ Hybridization, Fluorescence , Male , Nucleic Acid Hybridization , Prostatic Neoplasms/pathology , Reproducibility of Results , Translocation, Genetic/genetics , Tumor Cells, Cultured
20.
Surgery ; 124(6): 1094-8; discussion 1098-9, 1998 Dec.
Article in English | MEDLINE | ID: mdl-9854589

ABSTRACT

BACKGROUND: The set point for parathyroid hormone (PTH) secretion is increased in patients with primary hyperparathyroidism, possibly because of receptor defect(s). A decreased expression of calcium receptor (CaR) messenger ribonucleic acid (mRNA) and protein and a decreased expression of the putative calcium-sensing CAS (gp330/megalin) protein have been demonstrated in parathyroid adenomas. METHODS: Expression of CAS mRNA was studied in matched pairs of adenomas and adenoma-associated biopsy specimens from normal parathyroid glands from 15 patients with sporadic primary hyperparathyroidism. Cryostat sections were hybridized with an oligonucleotide complementary to CAS mRNA, rinsed, air dried, and exposed to x-ray film for semiquantification of radioactivity. RESULTS: Expression of CAS mRNA in the adenomas was lowered significantly to 25% (median; range 9% to 80%) of that of the corresponding biopsy specimens of normal parathyroid glands. No correlation was seen between CAS mRNA in the adenoma and preoperative serum calcium levels, PTH level, or weight of the adenoma. The levels of CAS mRNA were significantly lower than those observed previously for CaR mRNA. There was no significant correlation between the levels of CAS and CaR mRNA. CONCLUSIONS: Lowered levels of receptors sensing extracellular calcium (CaR and CAS) probably contribute to the increased set point for PTH secretion in primary hyperparathyroidism.


Subject(s)
Adenoma/genetics , Calcium-Binding Proteins/genetics , Gene Expression Regulation, Neoplastic , Parathyroid Neoplasms/genetics , Proteins/genetics , RNA, Messenger/biosynthesis , Apoptosis , Cellular Apoptosis Susceptibility Protein , Humans
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