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Alu profiling of primary and metastatic nonsmall cell lung cancer.

The metastatic potential of nonsmall cell carcinoma of lung (NSCLC), is currently recognized post factum, when lymph nodes or distant organs are already involved. Our ability to determine which tumors have acquired metastatic potential could help direct therapy to be more aggressive or less aggressive based upon this information. Evaluation of microsatellite instability via detection of LOH at specific loci may be useful in identifying specific markers and/or genes associated with this process. We examined Alu insertional elements as a potential marker of genetic changes associated with the metastatic potential of NSCLC. We analyzed archived, paraffin embedded tissue from 20 proven cases of NSCLC. DNA was extracted from 10 micron paraffin sections and amplified using an Alu PCR protocol. This technique does not examine specific loci but rather results in a banding profile of cellular genomic DNA. Informative allelic banding patterns, noted as differences between primary and metastatic lesions from the same patient, were observed in five of six cases (83%) with intrapulmonary metastases and in only nine of 14 (64%) cases with extrapulmonary metastases. Multiple genomic changes were detected in metastatic tumor cells as compared to normal lung tissue or primary lung tumor tissue. It appears that Alu profiling may be useful in the detection of metastatic vs primary lesions, and this technique may offer a method for identifying novel genes responsible for tumor progression and metastases.

Adult↗

The diagnostic value of flow cytometric DNA measurements in follicular tumors of the thyroid gland.

Flow cytometric analysis of DNA content was performed on single or multiple samples from 34 thyroidectomy specimens. There were 29 thyroids with diploid DNA content, comprising 15 nonneoplastic lesions, 5 follicular adenomas, 1 medullary carcinoma, and 8 papillary carcinomas. Aneuploid DNA pattern was observed in five cases, including one metastatic mammary carcinoma. The initial histologic diagnoses in the remaining four aneuploid thyroid lesions were follicular carcinoma in one and follicular adenoma in three. The abnormal DNA pattern in the three follicular "adenomas" prompted a review of their aspiration cytologic and histologic features. The fine-needle aspiration biopsy was performed in two of the three cases and showed evidence of a follicular neoplasm with significant nuclear atypia in both. Histologic review of the three lesions led to a modified diagnosis of noninvasive low grade follicular carcinoma in all three. Flow cytometric analysis of DNA content may prove to be a highly useful adjunct in the evaluation of follicular thyroid tumors. Long-term clinical follow-up is warranted to document the clinical significance of these observations.

Adenocarcinoma↗

Cytogenetic analysis of the mature teratoma and the choriocarcinoma component of a testicular mixed nonseminomatous germ cell tumor.

We karyotyped two histologically distinct components with different metastatic behavior of a testicular nonseminomatous germ cell tumor. The two components showed an almost identical chromosomal pattern. These almost identical karyotypes of the two components with different metastatic potential suggest that the difference in biologic behavior might result from subtle differences (on microscopic or submicroscopic level) in chromosomal pattern or that these differences are predominantly epigenetically determined and depend primarily on the lineage of differentiation of the tumor component. Trophoblastic differentiation results in an aggressive, angioinvasive tumor but in development of teratoma in a tumor with low malignant potential.

Adult↗

Immunohistochemical analysis of type IV collagenase expression in prostatic hyperplasia and adenocarcinoma.

The 72-kDa Type IV collagenase (T4C) is a matrix metalloproteinase, the expression of which may be important in normal basement membrane metabolism. The production of T4C by malignant cells has been linked to their invasive and metastatic potential in several tumor systems. The pattern of T4C immunoreactivity was assessed in formalin-fixed, paraffin-embedded prostatic tissue using polyclonal, monospecific antibodies. Basal cells in normal and hyperplastic epithelium demonstrated slight to moderate cytoplasmic immunoreactivity, while secretory epithelial cells showed generally weaker immunostaining. In 35 of 35 cases of primary adenocarcinoma and five of five cases of metastatic adenocarcinoma in lymph nodes, the majority of malignant cells showed strong immunoreactivity. Similar strong immunostaining was also seen in foci of prostatic intraepithelial neoplasia. The high level of expression of T4C in prostatic adenocarcinoma and its metastases suggests this metalloproteinase may play a role in determining the invasive and metastatic properties of this tumor. The enhanced immunoreactivity in prostatic intraepithelial neoplasia suggests the induction of T4C may be an early event in the development of the invasive phenotype. The expression of T4C in benign epithelial cells may be a manifestation of its putative physiological role in basement membrane metabolism.

Adenocarcinoma↗

Malignant insulinoma: spectrum of unusual clinical features.

BACKGROUND: Malignant insulinoma occurs in a few patients with insulinoma. Due to the small sample of patients, there are little data regarding their clinical manifestation as well as the preferred treatment modalities. The aims of the current study were to summarize the National Institutes of Health experience during the last two decades and to conduct a critical review of the current literature. METHODS: The authors identified 10 patients with metastatic insulinoma. RESULTS: The patients presented with four patterns of clinical behavior. First, four patients presented with lymph node metastasis and, after surgical excision, maintained a prolonged tumor-free survival. Second, four patients presented with metastatic disease to the liver, which appeared years after the initial diagnosis and presumed curative surgery. Third, one patient presented with a large alpha-fetoprotein-secreting liver mass. Finally, 9 of the 10 patients had a prolonged survival. Various treatment modalities were used to control hypoglycemia. Short-term benefits were most often achieved with embolization and diazoxide. Less successful modalities included radiofrequency ablation, radical debulking surgery, verapamil therapy, octreotide therapy, and chemotherapy. CONCLUSIONS: The current study, as well as others, suggested that metastatic insulinoma may have a variable natural history. After the initial surgical resection, the biology of the tumor, rather than any treatment modality, was most likely the major determinant of long-term survival.

Adult↗

Histochemical and biochemical aspects of lectin binding glycoproteins in primary and metastatic adenocarcinoma in the lung.

It is often difficult to differentiate between primary adenocarcinoma and metastatic adenocarcinoma in the lung. At least three lectins-peanut treated with neuraminidase, Ricinus communis-I and succinylated wheat germ-showed highly positive glycoprotein binding pattern along the plasma membrane of primary and metastatic adenocarcinoma in the lung. In order to distinguish primary pulmonary adenocarcinoma from metastatic adenocarcinoma in the lung with the three lectins, 40 pulmonary adenocarcinomas, seven extrapulmonary metastatic adenocarcinomas, and 16 metastatic adenocarcinoma in the lung were studied. Extracts from fresh tissue homogenate, after fractionation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were transferred by Western blotting and stained with lectin, and the lectin-bound glycoconjugates were read by a densitometer. The results showed all three lectin-bound glycoconjugate of primary pulmonary adenocarcinoma cases to be concentrated at about 200 Kd, but those of normal lung tissue had different molecular weights. Metastatic adenocarcinoma in the lung had lectin-bound glycoconjugate of about 120 Kd as well as 200 Kd. The amount of lectin binding high molecular weight substance was lower in metastatic adenocarcinoma in the lung than in primary pulmonary adenocarcinoma, especially succinylated wheat germ (P < 0.001). Therefore, decreased expression of the lectin binding high molecular weight glycoprotein may be a biochemical change associated with metastatic potential.

Adenocarcinoma↗

Intracranial metastatic melanoma: correlation between MR imaging characteristics and melanin content.

OBJECTIVE: Preliminary reports based on limited numbers of cases have proposed that specific MR imaging patterns may permit a distinction between melanotic and amelanotic brain metastases in melanoma patients. The purpose of this study was to test this hypothesis by categorizing MR images obtained from a large series of patients and correlating the results with the percentage of melanin-containing cells in surgically resected metastases. SUBJECTS AND METHODS: The MR images of 30 patients with histologically proven intracerebral melanoma were reviewed retrospectively. Precontrast MR images were obtained with T1-weighted spin-echo sequences in axial and sagittal sections and with proton density-weighted and T2-weighted sequences in axial sections. After IV injection of gadopentetate dimeglumine (0.1 mmol/kg of body weight), T1-weighted images were obtained in axial and coronal sections. All patients had undergone gross total resection of the evaluated lesions. MR images of the metastases were reviewed and sorted into four groups on the basis of putative patterns: (1) melanotic pattern--hyperintense in relation to cortex on T1-weighted images, hypointense in relation to cortex on T2-weighted images, and isointense or hyperintense in relation to cortex on proton density-weighted images; (2) amelanotic pattern--hypointense or isointense in relation to cortex on T1-weighted images and hyperintense or isointense in relation to cortex on T2-weighted and proton density-weighted images; (3) indeterminate, or mixed, pattern--MR imaging characteristics that did not conform to those of one of the first two categories; and (4) hematoma pattern--MR imaging features that exhibited only hematoma characteristics. Tissue sections from all evaluated lesions were independently reviewed by a neuropathologist (G.N.F.), and the percentage of melanin-containing tumor cells in each resected metastatic lesion was estimated. The MR imaging data and histologic data were then compared to assess the predictive value of the MR imaging patterns. RESULTS: Forty-two metastatic lesions were identified and categorized by MR imaging pattern as follows: 10 melanotic, 11 indeterminate (mixed), 16 amelanotic, and five hematoma. Correlation with histologic findings revealed that a majority (7/10) of lesions that exhibited a melanotic MR imaging pattern had more than 10% melanin-containing cells, over half (9/16) of lesions that exhibited an amelanotic MR imaging pattern contained histologically identifiable melanin (but always in less than 10% of cells), and lesions that exhibited a mixed MR imaging pattern were either amelanotic or contained less than 10% melanotic cells. Conversely, a majority of lesions containing more than 10% melanotic cells (7/8) demonstrated the typical melanotic MR imaging pattern, lesions with less than 10% melanin-containing cells exhibited a variety of MR imaging patterns, and only about half of patients with amelanotic lesions (6/13) showed the characteristic amelanotic MR imaging pattern. For five lesions, potentially informative imaging data on melanin content was obscured by histologically documented hematoma formation. CONCLUSION: Only a minority of melanoma metastases have the anticipated MR imaging findings of melanotic melanoma, which consist of high signal intensity relative to that of cortex on T1-weighted images and low signal intensity relative to that of cortex on T2-weighted images. Of tumors that do exhibit this melanotic pattern, the majority have more than 10% melanin-containing cells. The putative MR imaging pattern for amelanotic melanoma is nonspecific, as over half of tumors with this pattern contain melanin.

Adult↗

Hemodynamic considerations in organ and tissue patterning of B16 melanoma systemic metastasis and colonization.

Several populations of the mouse B16 melanoma that are highly metastatic from subcutaneous transplants but differ in growth characteristics were compared with regard to systemic site patterning of visible metastasis, as well as colonization effected by intracardiac injection of tumor cells. In all cases, metastasis proceeded in two stages, initially to the lungs and secondarily from lung metastases to systemic sites. The relative ranking of systemic site involvement by secondary-stage metastasis was basically similar for all tumor cell populations; the overall hierarchy was: kidneys greater than brain greater than adrenals and ovaries greater than pancreas greater than mesentery. Colonization patterns resulting from intracardiac injection were also generally comparable but differed from metastasis patterning in that the kidneys and brain were poorly colonized while the bones were frequent sites of colonization. Enumeration of fluoresceinated tumor cells or microbeads trapped in various sites following intracardiac injection revealed a ranking of initial involvement that differed markedly from colony formation. These results indicate that the hemodynamics of blood flow is not a critical determinant of colonization patterning. Based on the colonizing behavior of microbead-bound tumor cells, the frequent metastatic involvement of the kidneys and brain appears to result from selective trapping of large multicell tumor emboli within arteries in those organs. The occurrence of metastasis in other systemic sites is, like colonization, not readily explained by hemodynamics.

Animals↗

p53 mutations in prostate cancer bone metastases suggest that selected p53 mutants in the primary site define foci with metastatic potential.

PURPOSE: This study was undertaken to establish the pattern of specific p53 gene mutations in prostate cancer within primary tumors and distant metastases. MATERIALS AND METHODS: We performed polymerase chain reaction-single-strand conformation polymorphism and sequencing analyses of p53 exons 5-8 in DNA extracted from 22 formalin-fixed, paraffin-embedded tissues from 17 patients. Samples from three patients included specimens from primary and metastatic sites (paired specimens). RESULTS: G:C-to-A:T transitions were the most common point mutations (64%). Six (55%) of 11 G:C-to-A:T transitions occurred at CpG dinucleotides in five hot-spot codons (175, 245, 248, 273, and 282). Sequencing analysis of the paired samples revealed that two of the three pairs had the same mutation in both. Sequencing analysis of DNA from a different area of one of the primary tumors revealed a different mutation in the p53 gene. CONCLUSIONS: Our results suggest that specific p53 mutations participate in the progression of human prostate cancer. These findings support those of others that indicate that the primary cancer is heterogeneous and clonal expansion occurs during the progression of clinically detectable prostate cancer. Our data also imply that p53 mutations at the primary site may be predictive of metastases.

Aged↗

Parallel analysis of sporadic primary ovarian carcinomas by spectral karyotyping, comparative genomic hybridization, and expression microarrays.

Analysis of ovarian carcinomas has shown that karyotypes are often highly abnormal and cannot be identified with certainty by conventional cytogenetic methods. In this study, 17 tumors derived from 13 patients were analyzed by a combination of spectral karyotyping (SKY), comparative genomic hybridization (CGH), and expression microarrays. Within the study group, a total of 396 chromosomal rearrangements could be identified by SKY and CGH analysis. When the distribution of aberrations was normalized with respect to relative genomic length, chromosomes 3, 8, 11, 17, and 21 had the highest frequencies. Parallel microarray expression studies of 1718 human cDNAs were used to analyze expression profiles and to determine whether correlating gene expression with chromosomal rearrangement would identify smaller subsets of differentially expressed genes. Within the entire set of samples, microarray expression analysis grouped together poorly differentiated tumors irrespective of histological subtype. For three patients, a comparison between genomic alterations and gene expression pattern was performed on samples of primary and metastatic tumors. Their common origin was demonstrated by the close relationship of both the SKY and CGH karyotypes and the observed profiles of gene expression. In agreement with the pattern of genomic imbalance observed for chromosome 3 in ovarian cancer, the relative expression profile with respect to a normal ovary exhibited a contiguous pattern of reduced expression of genes mapping to the 3p25.5-3p21.31 and increased expression of genes from 3q13.33-3q28. This study demonstrates that SKY, CGH, and microarray analysis can in combination identify significantly smaller subsets of differentially expressed genes for future studies.

Adult↗

Spectral parameter imaging for detection of prognostically significant histologic features in uveal melanoma.

Specific extracellular matrix patterns in uveal melanoma are associated with metastatic risk. The laminin-rich composition and dimensions (on the order of a wavelength or less) of these structures suggest that acoustic backscatter might be affected by their presence. In this study, 10-MHz radiofrequency (RF) ultrasound (US) data were acquired before surgical removal of 117 eyes with uveal malignant melanoma. Histologic sections were evaluated for the presence of matrix patterns and acoustic backscatter was characterized using calibrated spectrum analysis. Statistical correlations between acoustic and histologic patterns were determined and linear discriminant analysis (LDA) and radial basis networks (RBN) were used to develop classification models for histologically based risk groups. Statistically significant correlations were found between acoustic parameters and the presence of histologic matrix-rich patterns. Retrospective classification accuracies of 74.4% and 78.6% were obtained with LDA and RBN, respectively. Leave-one-out analyses indicated estimated predictive accuracies of 71.8% and 75.0% for LDA and RBN, respectively.

Extracellular Matrix↗

[Cause of increased focal uptake in the cervical vertebrae in skeletal scintigraphic routine studies].

Focal uptake was seen in the cervical vertebral column of 61 patients in the course of routine bone scintigraphy. These focal increases were subjected to close analysis. In 60 of these patients (98.4%) degenerative lesions became manifest as the cause of the scintigraphic finding already by the bone scintigraphy pattern (paravertebral localisation in the posterior projection and dorsal localisation in the lateral. Two typical examples of metastatic changes are presented to facilitate differentiation. The bone scintigraphic pattern in arthrosis of the intervertebral joint is so typical that x-ray control examinations to confirm the diagnosis can be omitted in many cases.

Aged↗

Diverse signaling pathways through the SDF-1/CXCR4 chemokine axis in prostate cancer cell lines leads to altered patterns of cytokine secretion and angiogenesis.

The establishment of metastatic bone lesions in prostate cancer (CaP) is a process partially dependent on angiogenesis. Previously we demonstrated that the stromal-derived factor-1 (SDF-1 or CXCL12)/CXCR4 chemokine axis is critical for CaP cell metastasis. In this investigation, cell lines were established in which CXCR4 expression was knocked down using siRNA technology. When CaP cells were co-transplanted with human vascular endothelial cells into SCID mice, significantly fewer human blood vessels were observed paralleling the reductions in CXCR4 levels. Likewise, the invasive behaviors of the CaP cells were inhibited in vitro. From these functional observations we explored angiogenic and signaling mechanisms generated following SDF-1 binding to CXCR4. Differential activation of the MEK/ERK and PI3K/AKT pathways that result in differential secretion IL-6, IL-8, TIMP-2 and VEGF were seen contingent on the cell type examined; VEGF and TIMP-2 expression in PC3 cells are dependent on AKT activation and ERK activation in LNCaP and LNCaP C4-2B cells leads to IL-6 or IL-8 secretion. At the same time, expression of angiostatin levels were inversely related to CXCR4 levels, and inhibited by SDF-1 stimulation. These data link the SDF-1/CXCR4 pathway to changes in angiogenic cytokines by different signaling mechanisms and, suggest that the delicate equilibrium between proangiogenic and antiangiogenic factors may be achieved by different signal transduction pathways to regulate the angiogenic phenotype of prostate cancers. Taken together, our results provide new information regarding expression of functional CXCR4 receptor-an essential role and potential mechanism of angiogenesis upon SDF-1 stimulation.

Angiostatins↗

The microcirculation of choroidal and ciliary body melanomas.

The microcirculation of ciliary body and choroidal melanomas is remodelled into patterns. The presence of microvascular networks, composed of back-to-back loops that encircle microdomains of tumour, and parallel vessels with cross-linking, are associated with death from metastatic melanoma. The formation of these complex vascular patterns may result from reciprocal interactions between the tumour cell and the extracellular matrix, and pattern formation may reflect an invasive tumour cell phenotype. Ciliary body and choroidal melanomas are among the few forms of cancer treated before a pathologist assigns a grade to indicate whether tumour is likely to follow a benign or aggressive course. There is evidence to suggest that prognostically significant microcirculatory patterns may be detectable by non-invasive imaging techniques that may provide a substitute for biopsy to guide the clinical management of patients with these sight- and life-threatening tumours.

Choroid Neoplasms↗

Underpants-pattern erythema: a previously unrecognized cutaneous manifestation of extramammary Paget's disease of the genitalia with advanced metastatic spread.

BACKGROUND: A previously unrecognized erythema was noted in 6 patients with extramammary Paget's disease (EMPD) of the genitalia with metastases. It was distinct from essential erythema of EMPD and has not been reported before. OBJECTIVE: Our purpose was to describe the clinical and histopathologic features of the erythema and clarify its pathogenesis and prognostic importance. METHODS: A review of the clinical records and histologic materials of 6 cases was made focusing on the clinical features of this unique erythema as well as the course and prognosis of the patients. RESULTS: The erythema first appeared in the inguinal region and extended centrifugally to an area normally covered by underpants. Histopathologic examination showed lymphatic infiltration by cancer cells of the skin. All of the patients with this "underpants-pattern erythema" died of the disease, and the mean survival period after its appearance was 13 months. CONCLUSION: Underpants-distribution erythema may be a sign of advanced stage EMPD.

Aged↗

The pattern of malignant tumours of the liver in a tertiary health institution in Nigeria.

This study aims at establishing the histopathological pattern of liver cancers reported in the Department of Pathology, University of Ilorin Teaching Hospital between January 1979 and December 1996. Histopathological slides and duplicate copies of reports were retrieved and re-examined while fresh sections were processed from original paraffin blocks when necessary. Liver cancers constituted 56.6% of all liver biopsies. Hepatocellular carcinoma, which was commonest in the fifth decade, occurred in 64.1%, while metastatic tumours, which were commonest in the sixth decade, accounted for 24.7%. Primary sites could only be determined in 5 out of the 35 metastatic tumours. The age range of patients with Burkitt's lymphoma was 6-17 years with an average of 10.8 years and a male/female ratio of 3:2. Overall, there was preponderance of hepatocellular carcinoma in males while metastatic tumours were commoner in females. The pattern of Burkitt's lymphoma demonstrated in this study agrees with what generally obtains for endemic Burkitt's lymphoma in this environment.

Adenocarcinoma↗

Retreating recurrent breast cancer with the same CMF-containing regimen used as adjuvant therapy. The International Breast Cancer Study Group.

Breast cancer metastases appearing soon after adjuvant chemotherapy (within 12 months of its completion) are usually resistant to retreatment with the same cytotoxic agents, while relapses occurring later (beyond 12 months) regress when rechallenged with the same agents, showing similar response rates observed in non-pretreated patients with advanced disease. The International Breast Cancer Study Group (IBCSG) prospectively explored the efficacy of retreatment for patients upon relapse using the same therapy administered during the adjuvant programme. 87 patients previously treated with an adjuvant CMF (cyclophosphamide, methotrexate, 5-fluorouracil) combination chemotherapy (with or without the addition of low-dose prednisone and tamoxifen), who had measurable first breast cancer relapse, usually after at least 6 months of completion of the adjuvant treatment, were treated with CMF. Pretreatment consisted of 1-3 CMF courses in 27 patients and 4 or more courses in 60 patients. 17 patients were retreated with additional tamoxifen or had tamoxifen stopped at relapse. The data of these patients are shown separately. 47 of the 86 fully evaluable patients (55%) had an objective response, which was complete in 25 (29%). The dominant metastatic type and the number of involved sites were the most important factors influencing response to retreatment. Patients with soft tissue metastases had a high response rate (36/52, 69%) compared with those who had visceral involvement (9/24, 38%) or those with bony disease (2/10, 20%) (P = 0.002). In conclusion, response rates to retreatment with CMF were similar to those expected in a non-pretreated population. The patterns of relapse and the number of metastatic sites were the most important factors predicting response to retreatment, while treatment-free interval (usually longer than 6 months due to the study design) did not influence response rates. This study supports the hypothetic effectiveness of late reintroduction of adjuvant cytotoxic therapy (prior to evidence of systemic relapse), upon which several current trials are based.

Adult↗

Pathogenesis of primary central nervous system lymphoma: invasion of malignant lymphoid cells into and within the brain parenchyme.

The pattern of invasion of lymphoid cells to the central nervous system (CNS) was analyzed for malignant lymphocytes in 19 primary CNS lymphomas (PCNSL) and six intracerebral metastatic lymphomas, and for reactive lymphocytes in four encephalitides and three astrocytomas. The identical spreading pattern in both primary and metastatic lymphomas suggests that even in the so-called primary CNSL the malignant transformation has occurred outside the CNS. The compact perivascular cuffs of both malignant and reactive lymphocytes were never seen around the smallest capillaries, and they were most common around vessels larger than 15 microns in diameter. Perivascular lymphocytes resided within the reticulin network, which was immunopositive for collagen type III and IV, laminin and fibronectin. These findings imply that lymphocytes extravasate at the level of arterioles and venules and spread along the enlarged perivascular space. When the outer boundary of the perivascular network was broken, malignant lymphocytes spread diffusely into the CNS parenchyme; a pattern which is different from that of other CNS metastases. The widespread immunopositivity for the homing cell adhesion molecule CD44 in the CNS vessels and parenchyme, especially in the white matter which is the predilection site of PCNSL, suggest that this adhesion molecule and its ligands participate in spreading of malignant lymphocytes within the CNS parenchyme.

Adult↗