Torill Sauer

Detection of Isolated Tumor Cells in Bone Marrow Is an Independent Prognostic Factor in Breast Cancer

PURPOSE This study was performed to disclose the clinical impact of isolated tumor cell (ITC) detection in bone marrow (BM) in breast cancer. PATIENTS AND METHODS BM aspirates were collected from 817 patients at primary surgery. Tumor cells in BM were detected by immunocytochemistry using anticytokeratin antibodies (AE1/AE3). Analyses of the primary tumor included histologic grading, vascular invasion, and immunohistochemical detection of c-erbB-2, cathepsin D, p53, and estrogen receptor (ER)/progesterone receptor (PgR) expression. These analyses were compared with clinical outcome. The median follow-up was 49 months. RESULTS ITC were detected in 13.2% of the patients. The detection rate rose with increasing tumor size (P =.011) and lymph node involvement (P <.001). Systemic relapse and death from breast cancer occurred in 31.7% and 26.9% of the BM-positive patients versus 13.7% and 10.9% of BM-negative patients, respectively (P <.001). Analyzing node-positive and node-negative patients separately, ITC positivity was associated with poor prognosis in the node-positive group and in node-negative patients not receiving adjuvant therapy (T1N0). In multivariate analysis, ITC in BM was an independent prognostic factor together with node, tumor, and ER/PgR status, histologic grade, and vascular invasion. In separate analysis of the T1N0 patients, histologic grade was independently associated with both distant disease-free survival (DDFS) and breast cancer-specific survival (BCSS), ITC detection was associated with BCSS, and vascular invasion was associated with DDFS. CONCLUSION ITC in BM is an independent predictor of DDFS and BCSS. An unfavorable prognosis was observed for node-positive patients and for node-negative patients not receiving systemic therapy. A combination of several independent prognostic factors can classify subgroups of patients into excellent and high-risk prognosis groups.

Early detection of breast cancer based on gene-expression patterns in peripheral blood cells

IntroductionExisting methods to detect breast cancer in asymptomatic patients have limitations, and there is a need to develop more accurate and convenient methods. In this study, we investigated whether early detection of breast cancer is possible by analyzing gene-expression patterns in peripheral blood cells.MethodsUsing macroarrays and nearest-shrunken-centroid method, we analyzed the expression pattern of 1,368 genes in peripheral blood cells of 24 women with breast cancer and 32 women with no signs of this disease. The results were validated using a standard leave-one-out cross-validation approach.ResultsWe identified a set of 37 genes that correctly predicted the diagnostic class in at least 82% of the samples. The majority of these genes had a decreased expression in samples from breast cancer patients, and predominantly encoded proteins implicated in ribosome production and translation control. In contrast, the expression of some defense-related genes was increased in samples from breast cancer patients.ConclusionThe results show that a blood-based gene-expression test can be developed to detect breast cancer early in asymptomatic patients. Additional studies with a large sample size, from women both with and without the disease, are warranted to confirm or refute this finding.

Reduced expression of Claudin-7 in fine needle aspirates from breast carcinomas correlate with grading and metastatic disease.

Objective:  To study the immunocytochemical expression of the tight junction protein Claudin‐7 in smears from breast carcinomas and correlate with grading, nodal status, locoregional and distant metastases and the cellular cohesion.

Epidermal Growth Factor Receptor Inhibition Induces Trichomegaly

Case report. The epidermal growth factor receptor (EGFR) is important for normal skin development and function (1 /3). Binding of the ligands EGF or TGF-a induces dimerization of the receptor and activation of the intracellular tyrosine kinase, resulting in downstream activation of different signalling pathways including MAP-kinases. EGFR expression is upregulated in several malignancies and therapies targeting EGFR are now under development. Strategies include antibodies inhibiting ligand-binding and low molecular weight compounds inhibiting the intracellular tyrosine kinase. In this case report we describe growth of eyelashes (trichomegaly) in a 26 year-old female treated with irinotecan and Cetuximab (chimeric antibody against EGFR, Imclone) (see Fig. 1). The patient was diagnosed with colon cancer with liver metastases and received treatment with irinotecan (350 mg/m every 3 weeks) for several months. Owing to progressive disease, Cetuximab was added to the irinotecan treatment. As seen in most patients on irinotecan treatment, our patient developed alopecia grade II that was unaltered on the combined Cetuximab /irinotecan treatment. After the first week of treatment, the patient developed the typical acneiform erythematous rash commonly described in patients on Cetuximab and other EGFR inhibitors (4). She observed a marked increase in the length of her eyelashes and eyebrows after about 10 weeks of treatment. The photograph was taken after 20 weeks of treatment. Owing to the grade II acneiform erythematous rash, the patient began treatment with tetracycline (300 mg/day) starting in week 19. The patient had a clinical, radiological and biochemical response to Cetuximab treatment with a reduction in tumour size of 47% and plasma CEA levels were decreased from 432 to B/5 (normal range). A skin biopsy taken at 28 weeks after start of treatment showed histopathological findings as previously described in patients on EGFR inhibitors (4), with reduced Ki-67 expression compared to normal skin biopsies. There was no evidence of androgen-stimulated hair growth. Plasma sex hormone levels and elevated SHBG were in the range normally seen in young females taking contraceptive pills. The patient did not use any other medication. Similar trichomegaly was also observed in another female patient with metastatic colon cancer who was treated with Cetuximab monotherapy after progression on irinotecan treatment. Because of the marked and inconvenient increase in the length of her eyelashes, she resorted to using scissors to trim her eyelashes.

Relationship Between Intestinal Fibrosis and Histopathologic and Morphometric Changes in Consequential and Late Radiation Enteropathy

Intestinal fibrosis is a marked feature of late radiation enteropathy. This study assessed the time dose fractionation relationships of radiation-induced fibrosis in order to elucidate possible pathogenetic mechanisms. In 290 male Sprague-Dawley rats, a loop of small bowel was transposed to the left side of the scrotum. Three weeks later, the transposed segment was irradiated with either single dose or various fractionated regimens. The animals were observed for radiation-induced intestinal complications and killed in groups, 2 and 26 weeks after completion of irradiation. A semiquantitative histopathologic radiation injury score, morphometry of the submucosa, submucosal arterioles, intestinal surface area, and relative collagen content were used as endpoints. Fibrosis, measured by collagen assay and radiation injury score, increased with total dose, increasing fraction size and reduction in overall treatment time. This paralleled the results of morphometric assessment of mucosal surface area. Differences in vascular morphometry were only statistically significant in response to changes in total dose and fraction size and not with changes in overall treatment time. We conclude that fibrosis increases with increasing observation time, increasing fraction size, increasing total dose, and reduction of interfraction interval. Consequential injury, occurring as a result of disruption of mucosal integrity, seems to be an important mechanism for development of intestinal fibrosis. In contrast, vascular injury is relatively independent of this mechanism.

LATE CHANGES FOLLOWING SINGLE DOSE ROENTGEN IRRADIATION OF RAT SMALL INTESTINE

In female Wistar rats a 10 cm long exteriorized mid small intestinal segment was roentgen irradiated with 17, 19, 21 and 23 Gy as single exposures. Animals were killed in groups of 3 at intervals of 6 weeks from 2 to 50 weeks following irradiation. Irradiation injury was assessed using 8 macroscopic and histopathologic parameters, and an injury score for each animal was calculated. The parameters used were divided in 2 subgroups, early and late alterations, showing different types of development. The score for the early alterations decreased from 2 to 20 weeks following irradiation and then remained constant. The late alterations increased and seemed to stabilize about 8 weeks following irradiation. After the initial 20 weeks there was no progression of irradiation injury.

Expression pattern of adhesion molecules (E-cadherin, α-, β-, γ-catenin and claudin-7), their influence on survival in primary breast carcinoma, and their corresponding axillary lymph node metastasis†

Reduced intercellular adhesion is implicated in the development of metastasis. This study investigates the expression of intercellular adhesion molecules (E‐cadherin, α‐, β‐, γ‐catenin and claudin‐7) and their influence on survival in primary breast carcinomas and corresponding axillary lymph node metastases (ALNM), and evaluates associations between them and with clinicopathological factors. The expression of adhesion molecules was analyzed immunohistochemically in tissues from 196 patients with primary invasive breast carcinomas and their nodal metastases (174 ductal and 22 lobular types). The expression was evaluated using semi‐quantitative scoring of the intensity and proportion of immunoreactivity. All five adhesion proteins showed significantly reduced expression in primary ductal carcinomas with re‐expression in ALNM (p<0.001). In uni‐ and multivariate analyses, the expression of E‐cadherin in the primary tumours was a significant predictor of disease‐free survival and distant disease‐free survival. Thus, abnormal E‐cadherin expression in the primary invasive breast carcinoma seems to be an independent prognostic biomarker in predicting a shorter survival in node‐positive breast cancer patients. The results indicate that abnormal expression of the adhesion molecules in the primary tumours with re‐expression in corresponding nodal metastases is a common event in breast ductal carcinomas and may play a central role in establishing metastasis.

TOLERANCE OF RAT SMALL INTESTINE TO LOCALIZED SINGLE DOSE AND FRACTIONATED IRRADIATION

The tolerance of rat small intestine to localized single-dose and fractionated irradiation was assessed. In 168 rats, bilateral orchiectomy was performed and a loop of small intestine was transposed to the left part of the scrotum. Beginning 3 weeks postoperatively, single dose (18-24 Gy) or fractionated (4.2 Gy or 5.6 Gy per fraction) x-irradiation was delivered to the transposed intestine. The animals were observed for complications, and groups of animals were killed 2 and 26 weeks after completion of irradiation for assessment of injury. Mortality (i.e. the occurrence of lethal intestinal complications) and a semiquantitative histopathologic scoring system were used as endpoints to assess the degree of radiation injury. The most frequent intestinal complications were enterocutaneous fistula formation and intestinal obstruction. Logistic regression analysis ov complications data was used to estimate LD50 values and the alpha/beta ratio. There was good correlation between histopathologic scores and the incidence of lethal complications. The estimated LD50 values were 22.1 +/- 0.5 Gy, 37.0 +/- 4.4 Gy and 51.0 +/- 5.3 Gy for the single dose regimen and the fractionated regimens of 5.6 Gy and 4.2 Gy respectively. The estimated alpha/beta ratio was 10.7 +/- 2.4 Gy. The goodness of fit of the linear-quadratic isoeffect model to our data was satisfactory. Our results indicate that acute mucosal damage may be pathogenetically involved in the development of intestinal complications.

An increased risk of cervical intra-epithelial neoplasia grade II-III among human papillomavirus positive patients with the HLA-DQA1*0102-DQB1*0602 haplotype: A population-based case–control study of Norwegian women

Several recent studies have reported different associations between HLA specificities and human papillomavirus (HPV)‐associated disease of the cervix. We report the distribution of DQA1 and DQB1 genes and HPV infection in a population‐based case–control study including 92 patients with histologically verified cervical intraepithelial neoplasia grade II‐III (CIN II‐III) (thus including moderate and severe dysplasia and carcinoma in situ) and 225 control subjects. We found an overrepresentation of the DQA1*0102‐DQB1*0602 haplotype among HPV‐positive cases compared with controls. The association was even stronger when comparing HPV‐16–positive cases with HPV‐16–positive controls. In addition, among HPV‐16–positive individuals, we observed a decreased frequency of DQA1*0102‐DQB1*0604 in cases compared with controls. We were not able to detect any association between CIN II‐III and DQB1*03. Compared with previous findings in cervical cancer, our data indicate that carrying the DQA1*0102‐DQB1*0602 haplotype gives an increased risk of developing CIN when infected with HPV‐16, without influencing progression to cancer. Int. J. Cancer 76:19–24, 1998.© 1998 Wiley‐Liss, Inc.

Demonstration of EGFR gene copy loss in colorectal carcinomas by fluorescence in situ hybridization (FISH): a surrogate marker for sensitivity to specific anti‐EGFR therapy?

Aims : To investigate EGFR gene copy number heterogeneity in colorectal carcinomas compared with copy number of chromosome 7 and immunohistochemical expression of the EGFR protein.