Chromosomal aberrations in transitional cell carcinoma that are predictive of disease outcome are independent of polyploidy

Objective To determine whether aneusomy for chromosomes 7, 9 and 17 (reported to predict recurrence in up to 65% of patients with superficial transitional cell bladder cancer and thus providing the opportunity for early and effective treatment) reflects specific genetic events on these chromosomes or merely wider unspecific genetic damage to the cell, e.g. that increased copy numbers for 7 and 17 reflect tumour polyploidy. Materials and methods The study comprised 25 primary tumours; 6 mu m thick sections from formalin-fixed and paraffin-embedded tumours were analysed. Chromosome copy numbers were determined by fluorescence in situ hybridization (FISH) using pericentromeric probes for chromosomes 7, 8, 9, 10, 11 and 17. A minimum of 200 nuclei per tumour area were scored by two independent observers. Results Eight of the 25 tumours examined (32%) showed no evidence of chromosomal abnormalities as detected by FISH for any chromosomes analysed. Twelve tumours (48%) showed abnormalities for one or two chromosomes, five tumours (20%) showed abnormalities for multiple chromosomes and one tumour showed abnormalities for all chromosomes analysed, suggestive of polyploidy. Conclusions Chromosomal abnormalities predictive of recurrence occur largely in the absence of other gross chromosomal lesions. In a small proportion of cases other chromosomes are affected, but this is almost always distinct from tumour polyploidy

Aneusomy of chromosomes 7 and 17 predicts the recurrence of transitional cell carcinoma of the urinary bladder

Objective To determine if changes in chromosome 7 and 17 copy number can be used to predict recurrence in patients with primary noninvasive (pTa) or superficially invasive (pT1) transitional cell carcinoma (TCC) of the urinary bladder. Patients and methods Tissue specimens for 129 tumours from 52 patients (38 men and 14 women) with pTa/pT1 TCC at first diagnosis were retrieved from pathology archives. All patient notes were accessed and disease outcome documented for superficial (pTa/ pT1) recurrence or progression to detrusor muscle invasion (greater than or equal to pT2). The rumours were examined for chromosomal copy number of chromosomes 7 and 17 using fluorescence in situ hybridization (PISH) with chromosome-specific probes. The copy number of chromosomes 7 and 17 was determined in interphase nuclei on intact 6 mu m tissue sections. Results Aneusomy of chromosomes 7 and 17 was detected in the index primary tumours of 10 of 32 (31%) patients with subsequent recurrent disease. No aneusomy for these chromosomes was detected in primary tumours from 20 patients with no detectable recurrence (P = 0.0082). The relative risk of recurrence was 3.62 times greater (95% confidence interval 1.6-8.1, Cox's multiple regression P = 0.0019) for patients with chromosomal aneusomy in primary TCC. Neither stage nor grade of the primary tumours was associated with recurrence in these patients, nor was there a significant association with increased grade (G2/3) or stage (greater than or equal to pT2) at recurrence. Conclusion These results suggest that the measurement of aneusomy by FISH, using markers for chromosomes 7 and 17, predict recurrence in a subgroup of patients with pTa/pT1 tumours at presentation. This finding may offer a new objective and quantitative test for patients destined to recur

Vacuum-assisted biopsy and steroid therapy for granulomatous lobular mastitis: report of three cases.

We report the cases of three patients with granulomatous lobular mastitis (GLM), who were treated successfully with low-dose steroid therapy. Furthermore, the findings of our review of 271 patients reported in the literature suggest that steroid therapy is the treatment of choice for GLM

Subgross breast pathology in the twenty-first century

Subgross studies on breast morphology in cleared thick sections are informative, but lengthy protocols could clash with diagnostic timescales and interfere with immunohistochemistry (IHC) or molecular analysis. We sought to speed up staining and tissue clearing to improve compatibility with diagnostic needs without detriment to histology or other assays. Thick sections (2–3 mm) of normal human breast tissue and whole mouse mammary glands were stained with alum-carmine, aceto-carmine, Harris's haematoxylin, cresyl violet, neutral red, thionin, 4',6-diamidino-2-phenylindole (DAPI), ethidium bromide (EB), or propidium iodide (PI) and cleared in xylene (refractive index ν = 1.50), benzyl alcohol/benzyl benzoate (BABB; ν = 1.59), xylene–BABB (X-BABB), thiodiglycol (2,2′-thiodiethanol, ν = 1.52) or anethole (ν = 1.56). Tissue was then paraffin embedded for IHC for ER, PR, E-cadherin, CD31 or cytokeratin 7. Haematoxylin and alum-carmine are excellent non-fluorescent subgross stains giving strong nuclear staining and minimal background. DAPI and EB permeate thick sections poorly but PI penetrates well, with a high signal-to-noise ratio after clearing in BABB or X-BABB. Other clearing agents were less effective, including thiodiglycol. Anethole's unpleasant odour precluded further evaluation. All evaluated clearing agents preserved comparable immunoreactivity for all markers. PI is a promising stain for subgross breast studies, compatible with BABB clearing. The hope that the water-miscible thioalcohol, thiodiglycol, might accelerate tissue clearing was not realised. There is scope for further streamlining to make subgross techniques more acceptable in combined research/diagnostic settings

Still waiting for predictive biomarkers in Barrett's oesophagus

Barrett's oesophagus is important as a precursor of oesophageal adenocarcinoma via a metaplasia-dysplasia-carcinoma sequence. It is often detected on upper gastrointestinal endoscopy. In the absence of glandular dysplasia the risk of progression to cancer is low but ascertainment of dysplasia is not always straightforward. Sparse mucosal sampling may miss dysplasia, or reactive changes may be overinterpreted due to inter and intraobserver variation. Low-grade and even high-grade dysplasia do not necessarily progress, provided prevalent cancer has been rigorously excluded. This indeterminacy motivates an ongoing search for clinically useful predictive biomarkers. Although many genetic and epigenetic abnormalities have been associated with neoplastic progression in Barrett's mucosa no molecular tests have as yet been accepted into routine pathology practice. Challenges of assay definition remain and many marker studies lack statistical power or have other methodological flaws. Even where strong evidence of clinically relevant predictive value does exist (in the case of ploidy analysis by flow or image cytometry) adoption has been minimal, likely reflecting technological and possible reimbursement obstacles. Well designed multicentre studies are likely to be required to translate improved knowledge of Barrett's carcinogenesis into clinically significant progress on predictive testing, and will require a degree of cooperation not so far widely seen in the field

The role of B lymphocytes in breast cancer: a review and current status

Primary breast carcinomas are often associated with tumour-infiltrating lymphocytes and metastatic carcinoma cells in axillary lymph nodes make intimate contact with lymphocytic cells. The defensive role of these lymphocytes against breast cancer remains controversial despite several decades of investigation. The identification of human tumour antigens recognised by the autologous host has provided convincing evidence for immune recognition of the tumour. Medullary breast carcinoma is characterized by heavy B lymphocyte infiltration, but these tumourinfiltrating lymphocytes clearly fail to eradicate it and apparently also fail to contain its growth, as the prognosis of medullary breast carcinoma may be no better than that of the more common ductal breast carcinoma. On the other hand, the effectiveness of herceptin has demonstrated that antibodies against tumour-associated antigens are potentially potent agents in the treatment of breast cancer. It is therefore possible that neoplastic zones that elicit effective immunological attack might be destroyed before they are clinically significant. The purpose of this article is to review the role of tumour-infiltrating B lymphocytes in breast cancer, the development of monoclonal antibodies from these lymphocytes, and their possible uses in therapy. It is not our intent nor can we discuss all breast cancer antigens and antibodies against them, as the numbers are too large, nor the role of other types of immune cells such as cytotoxic T cells and natural killer cells

Antigen-driven clonal proliferation, somatic hypermutation and selection of B-lymphocytes infiltrating human ductal breast carcinoma

Infiltration of B lymphocytes into the tumor tissue of breast cancer patients is a common occurrence, but the role of these cells in the immune response to the tumor is unknown. Heavy B-cell infiltration in medullary breast carcinoma is well documented and associated with a more favorable prognosis, implying a positive role for the humoral immune response in elimination of tumor cells. Variable B-cell infiltration has also been detected in infiltrating ductal carcinomas of the breast, but little is known about the immunoglobulin gene repertoire of these tumor-infiltrating B lymphocytes and whether they are actively responding to a local stimulus or merely passive bystanders. We have therefore investigated the repertoire of B cells infiltrating four invasive ductal carcinomas. A group of 233 rearranged Ig V-H genes was amplified, cloned, and sequenced from microdissected foci of infiltrating B cells. B cells within individual foci were polyclonal, and most were highly mutated. Several foci expressed dominant sets of V genes derived from B-cell clones. Some of these were found in more than one lymphoid cluster, indicating that B cells had migrated into the surrounding tissue and seeded new clusters. Analysis of the pattern of mutations in clonally related sets of Ig V genes expressed by tumor-infiltrating B cells shows that these cells are undergoing antigen-driven proliferation, somatic hypermutation, and affinity maturation in situ

Chromosome 17 aneusomy is associated with poor prognostic factors in invasive breast carcinoma

Aberrations of chromosome 17 are common in breast cancer. Fluorescence in situ hybridization (FISH) enables gene or chromosome copy number to be assessed in situ in archival tissues and related to morphology and clinical outcome. In this study direct labeled DNA probes for the chromosome 17 alpha satellite and the HER2/neu gene were applied simultaneously to 5 micron sections of 214 formalin-fixed paraffin-embedded invasive primary breast carcinomas. A high proportion (54%) of invasive breast carcinomas displayed aneusomy of chromosome 17. Polysomy 17 correlated with multiple copies of HER2/neu (p=less than 0.001), but not with HER2/neu amplification. Eighty-six patients without HER2/neu amplification had aneusomy 17. Fifty-eight of the 86 patients that had aneusomy 17 had high HER2/neu copy number. Twelve patients with normal copy number for chromosome 17 had amplification of HER2/neu and 30 patients had amplification of HER2/neu with aneusomy 17. Aneusomy 17 was associated with grade 3 carcinoma (p=0.008), ER negativity (p=0.0032) and a Nottingham prognostic index of greater than 5.4 (p=0.039) but was not associated with survival by univariate analysis. In conclusion, the determination of chromosome 17 copy number should be incorporated in assessment of HER2/neu status, as this will give an accurate measure of amplification of HER2/neu and may also be helpful in determining suitability for breast carcinoma trials

Expression of telomerase RNA in oesophageal and oral cancer

Background: Telomerase is detectable in 85% of cancers, but is largely repressed in normal tissues. Human telomerase RNA (hTR) inhibition is a promising anti-cancer strategy, but requires differential expression between malignant and normal tissue.<br/><br/> Method: Archival paraffin sections from 48 oral squamous cell carcinomas (SCC) (23 floor of mouth, 25 tongue) and 56 oesophageal carcinomas (41 SCC, 15 adenocarcinomas) were evaluated for hTR expression using a radiolabelled riboprobe. Results were compared with expression in controls and adjacent histologically normal tissue. Statistical analysis was by the chi<sup>2</sup> test.<br/><br/> Results: hTR was detectable in 76% of oral SCC overall (floor of mouth 65%, tongue 88%, P = 0.61), and in 54% of oesophageal cancers (SCC 51%, adenocarcinoma 60%, P = 0.56). Detectable hTR expression was significantly more frequent in oral SCC than oesophageal SCC (P = 0.01). hTR expression was only detected in normal tissue at low levels in basal squamous epithelium. There was agreement of hTR expression between 8/9 surgically excised carcinomas and their initial diagnostic biopsies. <br/><br/>Conclusion: Tumour-specific hTR expression confirms hTR inhibition as a possible therapeutic strategy in some if not all oesophageal and oral cancers. Generally concordant hTR status between biopsy and resection suggest that biopsy may have a role in selecting candidates for telomerase inhibition therapy

Tumour specific regulation of telomerase RNA gene expression visualized by in situ hybridization

Maintenance of telomere structure by the ribonucleoprotein enzyme telomerase is considered central to the development of most human cancers, However, regulatory mechanisms governing telomerase expression during oncogenesis are largely unknown. We address potential tumour-specific regulation of telomerase RNA gene expression by RNA in situ hybridization to over 300 tumour samples of germ cell and epithelial origin, Twenty-six per cent of non-small cell lung cancers (NSCLC), expressed detectable levels of the telomerase RNA gene (hTR), and interestingly expression was almost confined to squamous carcinomas (41%), being rare in pulmonary adenocarcinomas and large-cell anaplastic carcinomas (P=0.006), Low frequency hTR expression was also associated with adenocarcinoma of the breast (13%), and ovary (17%), In comparison, hTR expression was detected in 43% of cervical cancers with no significant differences in frequency between squamous-cell carcinoma and adenocarcinoma or in transitions between intraepithelial neoplasia and invasive carcinoma, In contrast to the common epithelial cancers, the malignant cells in 73% of testicular germ-cell tumours (seminomas and teratomas), expressed hTR consistent with hTR expression in normal testicular germ cells, Differentiated tissues within ovarian germ cell tumours and in testicular teratomas lacked detectable hTR expression, These studies show that different tumour types have distinct patterns of hTR expression, which has implications for our understanding of mechanisms regulating telomerase activity and for targeting the telomerase RNA component as an anti-cancer therapy