Pattern of employment and associated factors in long-term lymphoma survivors 10 years after high-dose chemotherapy with autologous stem cell transplantation.

Background This study examined employment patterns and associated factors in lymphoma survivors treated with high-dose chemotherapy with autologous stem cell transplantation (HDT-ASCT) from diagnosis to a follow-up survey at a mean of 10 years after HDT-ASCT. Patients and methods All lymphoma survivors aged ≥18 years at HDT-ASCT in Norway from 1987 to 2008, and alive at the end of 2011 were eligible for this cross-sectional study performed in 2012/2013. Participants completed a mailed questionnaire. Job status was dichotomized as either employed (paid work) or not-employed (disability and retirement pension, on economic support, home-makers, or students). Results The response rate was 78%, and the sample (N = 312) contained 60% men. Mean age at HDT-ASCT was 44.3 and at survey 54.0 years. At diagnosis 85% of survivors were employed, 77% before and 77% after HDT-ASCT, and 58% at follow-up. Forty seven percent of the survivors were employed at all time points. The not-employed group at survey was significantly older and included significantly more females than the employed group. No significant between-group differences were observed for lymphoma-related variables. Fatigue, mental distress and type D personality were significantly higher among those not-employed, while quality of life was significantly lower compared to the employed group. Older age at survey, being female, work ability and presence of type D personality remained significantly related to being not-employed at survey in the multivariable analysis. Conclusions Our findings show that not-employed long-term survivors after HDT-ASCT for lymphoma have more comorbidity, cognitive problems and higher levels of anxiety/depression than employed survivors. These factors should be checked and eventually treated in order to improve work ability.

Dose-densified chemoimmunotherapy followed by systemic central nervous system prophylaxis for younger high-risk diffuse large B-cell/follicular grade 3 lymphoma patients: results of a phase II Nordic Lymphoma Group study.

BACKGROUND: Many patients with aggressive B-cell lymphomas and high clinical risk score still die of lymphoma after conventional R-CHOP chemoimmunotherapy. We hypothesized that intensified chemoimmunotherapy including systemic central nervous system (CNS) prophylaxis improves outcome and reduces the incidence of CNS-related events. PATIENTS AND METHODS: Inclusion criteria were age 18-65 years, primary diffuse large B-cell lymphoma or grade III follicular lymphoma without clinical signs of CNS disease and negative cerebrospinal fluid cytology, age-adjusted International Prognostic Index 2-3 and WHO performance score 0-3. Treatment consisted of six courses of R-CHOEP-14 followed by a course of high-dose cytarabine and a course of high-dose methotrexate. Primary end point was failure-free survival (FFS) at 3 years. RESULTS: A total of 156 eligible patients with a median age of 54 years (range 20-64) were included. Three toxic deaths were observed. Three-year overall survival (OS) and FFS rates (median observation time 52 months for survivors) were 81% and 65%, respectively. Seven patients experienced CNS relapse, all within 6 months. CONCLUSIONS: The results are promising with favorable 3-year OS and FFS rates, a low toxic death rate and a lower than expected number of CNS events. CNS progression might be further reduced by earlier CNS prophylaxis. CinicalTrials.gov. identifier NCT01502982.

Expression of DSG1 and DSC1 are prognostic markers in anal carcinoma patients.

BACKGROUND: Our purpose was to investigate if dysregulation of cell adhesion molecules could be linked to prognosis in squamous cell carcinomas (SCCs) of the anal region. METHODS: Protein expression of desmoglein-1 (DSG1), desmocollin-1 (DSC1) and E-cadherin was studied by immunohistochemistry in a cohort of 53 anal carcinoma patients treated by radiation alone or combined with 5-fluorouracil and mitomycin C. RESULTS: Univariate analyses identified, among others, negative membranous DSG1 staining (P=0.009), negative cytoplasmic DSC1 staining (P=0.012) and negative DSG1 (membranous)+negative DSC1 (cytoplasmic) staining (P=0.004) to be associated with improved cancer-specific survival (CSS). On multivariate analyses positive DSG1 (membranous)+DSC1 (cytoplasmic) staining (HR 6.95, P=0.044), large tumour size and lymph node metastases (HR 6.44, P=0.004) and radiation without chemotherapy (HR 6.73 P=0.004) were associated with worse CSS. On univariate analysis, improved disease-free survival was associated with negative membranous staining of DSG1 (P=0.047), and negative DSG1 (membranous)+negative DSC1 (cytoplasmic) staining (P=0.025), among others. CONCLUSION: Membrane negativity for DSG1 and cytoplasmic negativity for DSC1 are favourable markers for CSS in SCCs of the anal region.

Expression of cyclin D1a and D1b as predictive factors for treatment response in colorectal cancer.

BACKGROUND: The aim of this study was to investigate the value of the cyclin D1 isoforms D1a and D1b as prognostic factors and their relevance as predictors of response to adjuvant chemotherapy with 5-fluorouracil and levamisole (5-FU/LEV) in colorectal cancer (CRC). METHODS: Protein expression of nuclear cyclin D1a and D1b was assessed by immunohistochemistry in 335 CRC patients treated with surgery alone or with adjuvant therapy using 5-FU/LEV. The prognostic and predictive value of these two molecular markers and clinicopathological factors were evaluated statistically in univariate and multivariate survival analyses. RESULTS: Neither cyclin D1a nor D1b showed any prognostic value in CRC or colon cancer patients. However, high cyclin D1a predicted benefit from adjuvant therapy measured in 5-year relapse-free survival (RFS) and CRC-specific survival (CSS) compared to surgery alone in colon cancer (P=0.012 and P=0.038, respectively) and especially in colon cancer stage III patients (P=0.005 and P=0.019, respectively) in univariate analyses. An interaction between treatment group and cyclin D1a could be shown for RFS (P=0.004) and CSS (P=0.025) in multivariate analysis. CONCLUSION: Our study identifies high cyclin D1a protein expression as a positive predictive factor for the benefit of adjuvant 5-FU/LEV treatment in colon cancer, particularly in stage III colon cancer.

MicroRNA-15b is induced with E2F-controlled genes in HPV-related cancer.

BACKGROUND: MicroRNAs (miRNAs) are important regulators of cellular processes and are found to be deregulated in many cancers. We here analysed the miRNA expression in anal carcinomas. In a previous study, we found that our anal carcinoma tumours were divided into two groups based on the expression of E2F-regulated genes. Therefore, we searched for miRNAs that could reproduce this grouping. METHODS: A global screen of the miRNA population was performed using real-time quantitative PCR (RT-qPCR) array methods and differentially expressed miRNAs were identified. Real-time-qPCR was used to verify the expression levels of selected miRNAs and genes in a larger collection of biopsies. A siRNA-mediated knockdown of human papilloma virus (HPV)16 E7 in a cervical cell line was performed to assess the effect of E7 on miR-15b. RESULTS: The grouping of tumours into two groups based on the expression of E2F-controlled genes was confirmed in a larger collection of anal carcinoma tumours. The expression of miR-15b was shown to be highly correlated with that of five selected E2F-induced genes (CCNA2, CCNB1, CCNB2, MSH6 and MCM7). A knockdown of HPV16 E7 resulted in decreased levels of miR-15b in Ca Ski cells. CONCLUSION: MiR-15b expression correlates with E2F-regulated genes in anal carcinoma and appears to be part of the E2F-regulatory network.

Expression of EZH2 and Ki-67 in colorectal cancer and associations with treatment response and prognosis.

BACKGROUND: Enhancer of zeste homologue 2 (EZH2) is a member of the Polycomb group of genes that is involved in epigenetic silencing and cell cycle regulation. METHODS: We studied EZH2 expression in 409 patients with colorectal cancer stages II and III. The patients were included in a randomised study, and treated with surgery alone or surgery followed by adjuvant chemotherapy. RESULTS: EZH2 expression was significantly related to increased tumour cell proliferation, as assessed by Ki-67 expression. In colon cancer, strong EZH2 expression (P=0.041) and high proliferation (>or=40%; P=0.001) were both associated with better relapse-free survival (RFS). In contrast, no such associations were found among rectal cancers. High Ki-67 staining was associated with improved RFS in colon cancer patients who received adjuvant chemotherapy (P=0.001), but not among those who were treated by surgery alone (P=0.087). In colon cancers stage III, a significant association between RFS and randomisation group was found in patients with high proliferation (P=0.046), but not in patients with low proliferation (P=0.26). Multivariate analyses of colon cancers showed that stage III (hazard ratio (HR) 4.00) and high histological grade (HR 1.80) were independent predictors of reduced RFS, whereas high proliferation indicated improved RFS (HR 0.55). CONCLUSION: Strong EZH2 expression and high proliferation are associated features and both indicate improved RFS in colon cancer, but not so in rectal cancer.

Influence of TP53 gene alterations and c-erbB-2 expression on the response to treatment with doxorubicin in locally advanced breast cancer.

TP53 status [mutations, immunostaining, and loss of heterozygosity (LOH)], expression of c-erbB-2, bcl-2, and histological grading were correlated to the response to doxorubicin monotherapy (14 mg/m2) administered weekly to 90 patients with locally advanced breast cancer. Mutations in the TP53 gene, in particular those affecting or disrupting the loop domains L2 or L3 of the p53 protein, were associated with lack of response to chemotherapy (P = 0.063 for all mutations and P = 0.008 for mutations affecting L2/L3, respectively). Similarly, expression of c-erbB-2 (P = 0.041), a high histological grade (P = 0.023), and lack of expression of bcl-2 (P = 0.018) all predicted chemoresistance. No statistically significant association between either p53 immunostaining or TP53 LOH and response to therapy was recorded, despite the finding that both were associated with TP53 mutation status (p53 immunostaining, P < 0.001; LOH, P = 0.021). Lack of immunostaining for p53 despite mutation of the TP53 gene was particularly seen in tumors harboring nonsense mutations or deletions/splices (7 of 10 negative for staining compared with 4 of 16 with missense mutations). TP53 mutations (total/affecting L2/L3 domains) were associated with expression of c-erbB-2 (P < 0.001 for both), high histological grade (P = 0.001 and P = 0.025), and bcl-2 negativity (P = 0.003 and P = 0.002). TP53 mutations, histological grade, and expression of bcl-2 (but not LOH or c-erbB-2 expression) all predicted for relapse-free as well as breast cancer-specific survival in univariate analysis (Ps between <0.0001 and 0.0155), but only tumor grade was found to be predictive in multivariate analysis (P = 0.01 and P = 0.0007, respectively). Our data are consistent with the hypothesis that certain TP53 mutations predict for resistance to doxorubicin in breast cancer patients. However, the observation that the majority of patients with TP53 mutations affecting or disrupting the L2/L3 domains with LOH in addition (n = 12) obtained a partial response (n = 4) or stabilization of disease (n = 5) during chemotherapy suggests redundant mechanisms to compensate for loss of p53 function. Our findings are consistent with the hypothesis that other defects may act in concert with loss of p53 function, causing resistance to doxorubicin in breast cancers.

Expression and alternative splicing of c-ret RNA in papillary thyroid carcinomas.

Somatic rearrangements of the ret receptor tyrosine kinase have been consistently reported in papillary thyroid carcinomas (PTC). It is unclear whether the expression of wild-type c-ret may also be implicated in thyroid tumorigenesis. We studied ret mRNA expression in PTC from Norwegian patients. Using RT-PCR, wild-type ret mRNA was detected in all of 22 PTC and in a PTC cell line. c-ret mRNA was clearly overexpressed in PTC as compared to non-neoplastic thyroid tissue. Hybridization using ret exon DNA dot blot arrays and complex cDNA probes confirmed expression of ret RNA in thyroid biopsies. In accordance with the RNA data, Western immunoblotting showed evidence of wild-type Ret protein in PTC. Rearrangements generating the ret/PTC oncogenes co-existed with c-ret mRNA in PTC. Multiple alternative ret splicing variants were detected in PTC. Four novel ret splicing events were found in the region encoding the extracellular domain. The open reading frames of these transcripts were all in-frame with the Ret tyrosine kinase domain. In the central ret mRNA region encoding the cysteine-rich, transmembrane, and main tyrosine kinase domains, no evidence of alternative splicing was detected. Two alternative splice events were detected in the ret mRNA encoding the C-terminal part of Ret protein harboring tyrosine residues important for Ret signaling, excluding exon 19, or retaining intron 19, respectively. Ribonuclease protection assays confirmed the presence of ret alternative splicing events in thyroid biopsies. We conclude that in addition to ret/PTC rearrangements, wild-type c-ret mRNA and alternatively spliced ret transcripts are present in PTC. Transcriptional up-regulation and post-transcriptional mechanisms of c-ret RNA processing may contribute to differences in expression of Ret protein observed in PTC compared to non-neoplastic thyroid tissue.

Proton nuclear magnetic resonance spectroscopy of serum from patients with colorectal neoplasia.

Proton nuclear magnetic resonance (NMR) spectra of serum have been recorded from patients with colorectal neoplastic polyps, before and after treatment of colorectal cancer, in patients with advanced lung cancer, and also from healthy controls. Digitally defined NMR profiles of the methyl and methylene peaks were used as input for supervised principal component modelling. An unknown sample was classified according to its residual, i.e. the difference between the spectral pattern of the unknown and control group. There was a statistically significant difference between the mean residual in the untreated colorectal cancer group and in controls (P = 0.003). The sensitivity of detecting untreated colorectal cancer was only 20%. There were no stage-dependent differences between the residuals within the untreated colorectal cancer group. After curative surgery, four patients had recurrence of malignant disease without an increase in residual prior to recurrence. Patients with advanced malignant disease (lung cancer WHO stage IIIB and IV) had a highly significant difference in mean residual from that of controls, with a sensitivity of detecting cancer of 87.5%. This increase in residual could not be explained by increase in the level of serum triglyceride. NMR spectroscopy was not a useful diagnostic tool in patients with colorectal neoplastic polyps and cancer.

Hospitalization for lung disease in early childhood and asthma symptoms in young adulthood.

Longitudinal studies have reported an association between early childhood lung disease and adult respiratory disease. This issue has not been addressed in the Nordic countries. We studied the association between hospitalization for lung disease in early childhood and asthma in young adulthood in a Norwegian population sample, while estimating the attributable fraction of childhood hospitalization. A population-based survey in Bergen, Norway included a random sample of 4300 subjects aged 20-44 years, of whom 80% responded. The effect of hospitalization for lung disease before the age of 2 years on asthma in adulthood was analysed by logistic and polytomous logistic regressions, adjusting for related variables. Adjusted attributable fractions were estimated from these models. The risk for different measures of asthma was significantly increased in the 103 persons reporting childhood hospitalization (airways symptoms: OR from 1.9 to 2.9; asthma medication: OR = 2.8). The associations with airways symptoms were stronger in women (OR from 2.6 to 5.3) than in men (OR from 1.4 to 2.4). Given a causal association, adjusted attributable fractions showed that childhood lung disease causing hospitalization explained 4% of asthma symptoms. Early childhood hospitalization for lung disease was related to asthma symptoms in young Norwegian adults, more strongly in women than in men. Only a minor proportion of asthma symptoms in this age group could be related to hospitalization for lung disease in early childhood.

Dietary vitamin C intake is inversely related to cough and wheeze in young smokers.

We aimed to investigate whether dietary vitamin C intake, an important antioxidant, is inversely related to self-reported respiratory symptoms in young adults of a community. A random sample of 4300 subjects, aged 20-44 years, living in Bergen, Norway, received a postal questionnaire on respiratory symptoms; 80% responded. Vitamin C intake (mg per week) was estimated from a food-frequency questionnaire asking how often the subject, during the last year, had consumed units of orange juice, oranges, potatoes, carrots and tomatoes. Significant differences in the intake of vitamin C were observed across smoking categories with current smokers having the lowest intake, while there was no variation by gender, age or occupational dust exposure. Dietary vitamin C intake was in univariate analyses inversely related to "morning cough", "chronic cough", "wheeze" and "wheeze ever". After adjusting for gender, age, body mass index, "occupational exposure" pack-years as well as having and stratified on smoking habits in multiple logistic regression analyses, the relationship between dietary vitamin C intake and "cough" and "wheeze" tended to be associated to smoking. The odds ratio (OR) for "morning cough" was 0.68 (95% CI: 0.35-0.95), "chronic cough" OR 0.69 (95% CI: 0.47-1.04) and "wheeze ever" OR 0.75 (95% CI: 0.56-1.01) in current-smokers with dietary vitamin C intake in the upper (> or =395 mg/ week) vs. the lower (<209 mg/week) tertile. The OR for "wheeze" was 0.56 (95% CI: 0.35-0.88) in ex-smokers. The magnitude ofthese effects remained after excluding subjects with supplementary vitamin C intake (n=199) from the statistical analyses. Among young Norwegian adults, having a low prevalence of asthma and high prevalences of smoking-related respiratory symptoms, dietary vitamin C intake may act as an antioxidant and thereby reduce cough and wheeze in smokers having high oxidant stress.

Pseudoprogression in high-grade glioma.

Pseudoprogression is a treatment-related effect seen on imaging in high-grade glioma. Enhancement of gadolinium contrast on control MRI can be misinterpreted as tumor recurrence and is also difficult to distinguish from radiation necrosis. Pseudoprogression is seen in up to 30% after standard treatment for glioblastoma multiforme (GBM), which is radiotherapy concurrent with chemotherapy with temozolomide (TMZ) and adjuvant cycles of TMZ. In this article, the current literature on pseudoprogression in high-grade glioma is reviewed by searches in PubMed. We also present two clinical cases, one of which had medullary pseudoprogression. No articles on this subentity of pseudoprogression were found in PubMed. Standard MRI with gadolinium contrast cannot differentiate between pseudoprogression, tumor recurrence and radiation necrosis. More advanced imaging techniques are often not available. Pseudoprogression seems to be related to methylated promoter of the O(6)--methyl-guanine methyl transferase (MGMT) gene, which is associated with improved treatment effect. Discontinuation or change of therapy on the basis of misinterpretation of MRI as disease progression is thus unfortunate. MRI should be interpreted with caution the first 6 months after standard treatment of high-grade glioma. In a GBM patient with contrast enhancement on MRI but few or no new symptoms and/or stable steroid doses, treatment should be continued and control imaging performed after 2-3 months.

Gene expression reveals two distinct groups of anal carcinomas with clinical implications.

Human papillomavirus (HPV) is a major aetiological agent in anal carcinomas. We here present a study of global gene expression using microarray hybridisation in a collection of anal carcinoma biopsies. Quantitative PCR was used to verify expression of selected genes. All biopsies contained integrated DNA of human papillomavirus subtype 16 (HPV16) and expressed HPV16 E7 mRNA. No other subspecies of HPV were detected in these 13 biopsies as assessed by PCR amplification and DNA sequencing. Unsupervised cluster analysis, based on global mRNA expression, divided the tumour biopsies into two distinct groups. Cluster analysis based on a number of high-risk HPV and/or E2F-regulated genes reproduced this biopsy grouping, suggesting that integrated HPV16 substantially influenced global gene expression in approximately half the biopsies studied. The levels of HPV16 E7 mRNA were significantly different between the two groups, but with considerable overlap. Thus, influence on global gene expression could not be absolutely ascribed to the expression level of HPV16. To investigate whether this distinction in gene expression had prognostic impact, we studied protein expression in an independent cohort of 55 anal carcinomas not included in the microarray study of two differentially expressed candidate genes, minichromosome maintenance complex component 7 (MCM7) and cyclin-dependent kinase inhibitor 2A (CDKN2A or p16). HPV status was assessed by in situ hybridisation. There was a significant association between in situ staining for HPV E7 mRNA and immunostaining for CDKN2A (p16) and MCM7 protein. CDKN2A (p16) mRNA was found significantly differentially expressed between the two tumour groups. However, cluster analysis on genes directly regulated by CDKN2A (p16) could not reproduce this split of biopsies into two groups, suggesting that the transcriptional regulatory activity of CDKN2A in these biopsies is inhibited. Furthermore, protein expression of CDKN2A (p16) could not be associated with survival. MCM7 is directly regulated by E2F and induced by HPV, and its mRNA was found differentially expressed between the two tumour groups. High level of MCM7 protein was found to be associated with both improved relapse-free survival (RFS, P=0.02) and cancer-specific survival (CSS, P=0.03) in anal cancer patients treated with radiation with or without additional chemotherapy.

[Lung shock in connection with acute myocardial infarction. Adult respiratory distress syndrome, ARDS]. / Sjokklunge ved akutt hjerteinfarkt. Adult respiratory distress syndrome, ARDS.

We describe the case of 42 year-old male patient who developed adult respiratory distress syndrome (ARDS) during an acute myocardial infarction. He was treated with positive pressure ventilation for 23 days. Chest X-ray showed multiple, bilateral infiltrates for several weeks. ARDS is accompanied by serious and progressive hypoxemia. Early diagnosis is important. The therapeutic consequences involve positive pressure ventilation with increased inspired oxygen concentration and positive end-expiratory pressure. ARDS in patients with impaired myocardial function can be difficult to recognize. It is an alternative diagnosis to pneumonia and ordinary heart failure when patients are seen to be suffering serious hypoxemia.

Fish consumption and respiratory symptoms among young adults in a Norwegian community.

The aim of this study was to investigate the relationship between dietary fish consumption and self-reported respiratory symptoms among young adults. A random sample of 4,300 subjects, aged 20-44 yrs, living in Bergen, Norway, received a postal questionnaire on respiratory symptoms, of whom 80% responded. Mean fish consumption was assessed in a food-frequency questionnaire by asking how often the subject consumed units of fish (150 g) during the last year. Average fish consumption was 1.8 units x week(-1). Fish intake of <1 unit x week(-1) was reported by 24%, 41% reported consumption of 1 unit x week(-1) and 35% intake of >1 unit x week(-1). A high fish intake was significantly associated with increasing age after adjusting for smoking. Adjusted for smoking habits, the prevalence of "cough at night" and "chest tightness" showed a decreasing trend with increasing fish consumption (p<0.05), while such a trend for "wheeze" was demonstrated only in smokers (p=0.008 for interaction). In logistic regression models (adjusting for age, sex, body mass, smoking habits and occupational exposure) fish consumption (three categories) was not significantly associated with "wheeze", "chest tightness", "breathless at night" or "asthma attack", although the odds ratios (OR) were consistently less than 1 (except for "asthma attack"). Fish consumption was of borderline significance as a protective factor of "cough at night", OR = 0.86 (95% confidence interval: 0.76-0.97) but in stratified analyses only in smokers. Subjects reporting very high levels of fish consumption (>14 units x week(-1)) did not have lower prevalences of respiratory symptoms. In conclusion, among young Norwegian adults, with a relatively low prevalence of asthma and an overall high fish intake, fish consumption was not a significant predictor of four out of five respiratory symptoms.

In vitro toxicity of purified gluten peptides tested by organ culture.

Various subfractions of Frazer fraction III were separated by high-pressure liquid chromatography, and their toxicity in vitro (organ culture) was tested in comparison with alpha-gliadin using duodenal biopsies from 25 patients with active celiac disease and subtotal villous atrophy, 2 patients with partial villous atrophy, and 10 nonceliac controls. One dominating fraction, designated Frazer III-2-VIII, was purified by several steps of rechromatography. It was markedly toxic to duodenal explants from patients with active celiac disease. The mean enterocyte height after culture was 15.9 microns compared with 25.6 microns in gluten-free medium. This difference was statistically significant in all cases except one, in which the lowest concentration (110 micrograms) was used. The in vitro toxicity of Frazer III-2-VIII was comparable with the toxicity of alpha-gliadin in twofold to fivefold higher concentrations. No toxicity could be detected in nonceliac explants (mean enterocyte height, 25.7 vs. 24.9 microns in gluten-free medium). The N-terminal amino acid sequence was (Gln)-Ile-Gln-Val-Phe-Pro-Ser-Gly-Gln-Val-Gln-(Trp)-Pro-Gln-Gln-(Gln)-Gl n-Pro- Phe-Pro. This sequence was not homologous to previously reported sequences of toxic gluten peptides. By use of the SwissProt and GenEMBL databases, it was concluded that the peptide Frazer III-2-VIII is part of the gamma-gliadin fraction.

Expression of heregulins and associations with the ErbB family of tyrosine kinase receptors in papillary thyroid carcinomas.

Heregulin (Hrg) growth factors are natural ligands for ErbB3 and ErbB4. Because these receptors are involved in papillary thyroid carcinomas, we studied expression of Hrgs in fresh-frozen thyroid tissue and analyzed for possible coexpressions among the 4 members of the ErbB family of growth factor receptors and Hrgs in papillary carcinomas. Immunohistochemistry for the Hrg precursor isoform (134 biopsies from 101 patients) showed nuclear immunostaining in 83% of papillary carcinomas but not in normal thyroid tissue. Cytoplasmic immunopositivity for the Hrg precursor isoform was moderate or strong in 78% of papillary carcinoma specimens and weak in 13% of normal thyroid tissue samples. Western blot for the Hrg precursor isoform showed the expected protein band of approximately 70 kDa in papillary carcinomas, but not in non-neoplastic thyroid biopsies. Whereas weak cytoplasmic immunostaining for the mature Hrg alpha, beta1, and beta3, was present in 48, 38, and 51% of papillary carcinomas, respectively, normal thyroid tissue samples were negative. Hrg mRNA was present in both tumor and nontumor tissue, with evidence of increased mRNA expression in 5 of 12 papillary carcinomas. RT-PCR of hrg mRNA, with subsequent DNA sequencing, confirmed the presence of hrg alpha, beta1, beta2, and beta3 mRNA in papillary carcinomas. In 55 papillary carcinomas, increased cytoplasmic immunostaining of the ErbB2 and ErbB3 receptors was significantly associated with each other and with cytoplasmic epidermal growth factor receptor (EGFR) immunoreactivity, indicating a common regulatory mechanism. Cytoplasmic staining for Hrg beta3 was significantly associated with ErbB3 immunostaining, indicating this receptor as the cognate one. The overexpression and nuclear localization of the Hrg precursor isoform were not associated with the expression of ErbB-receptors. This may reflect an unknown mechanism of action, possibly independent of the ErbB receptor system.

Molecular portraits of human breast tumours.

Human breast tumours are diverse in their natural history and in their responsiveness to treatments. Variation in transcriptional programs accounts for much of the biological diversity of human cells and tumours. In each cell, signal transduction and regulatory systems transduce information from the cell's identity to its environmental status, thereby controlling the level of expression of every gene in the genome. Here we have characterized variation in gene expression patterns in a set of 65 surgical specimens of human breast tumours from 42 different individuals, using complementary DNA microarrays representing 8,102 human genes. These patterns provided a distinctive molecular portrait of each tumour. Twenty of the tumours were sampled twice, before and after a 16-week course of doxorubicin chemotherapy, and two tumours were paired with a lymph node metastasis from the same patient. Gene expression patterns in two tumour samples from the same individual were almost always more similar to each other than either was to any other sample. Sets of co-expressed genes were identified for which variation in messenger RNA levels could be related to specific features of physiological variation. The tumours could be classified into subtypes distinguished by pervasive differences in their gene expression patterns.