Antigen unmasking on formalin‐fixed, paraffin‐embedded tissue sections

Cattoretti, Giorgio; Pileri, Stefano; Parravicini, Carlo; Becker, M. H. G.; Poggi, Simonetta; Bifulco, Carlo; Key, Göran; D'Amato, Lucia; Sabattini, Elena; Feudale, Elisa; Reynolds, Fred H.; Gerdes, Johannes; Rilke, F

doi:10.1002/path.1711710205

Cited by 675 publications

(347 citation statements)

References 25 publications

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“…The IHC used to reveal the Bcl-2 protein is not always performed with the same antibody. Sometimes the protocol was performed without prior reaction of epitope unmasking on fixed issue (Cattoretti et al, 1993). To try to exclude technical biases, we performed subgroup analysis according to the most frequently used methods: IHC with antibody clone 100 and clone 124 (Figures 1 and 2).…”

Section: Discussionmentioning

confidence: 99%

Role of Bcl-2 as a prognostic factor for survival in lung cancer: a systematic review of the literature with meta-analysis

et al. 2003

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The role of the anti-apoptotic protein Bcl-2 in lung cancer remains controversial. In order to clarify its impact on survival in small and non-small cell lung cancer (NSCLC), we performed a systematic review of the literature. Trials were selected for further analysis if they provided an independent assessment of Bcl-2 in lung cancer and reported analysis of survival data according to Bcl-2 status. To make it possible to aggregate survival results of the published studies, their methodology was assessed using a quality scale designed by the European Lung Cancer Working Party (including study design, laboratory methods and analysis). Of 28 studies, 11 identified Bcl-2 expression as a favourable prognostic factor and three linked it with poor prognosis; 14 trials were not significant. No differences in scoring measurement were detected between the studies, except that significantly higher scores were found in the trials with the largest sample sizes. Assessments of methodology and of laboratory technique were made independently of the conclusion of the trials. A total of 25 trials, comprising 3370 patients, provided sufficient information for the meta-analysis. The studies were categorised according to histology, disease stage and laboratory technique. The combined hazard ratio (HR) suggested that a positive Bcl-2 status has a favourable impact on survival: 0.70 (95% confidence interval 0.57 -0.86) in seven studies on stages I -II NSCLC; 0.50 (0.39 -0.65) in eight studies on surgically resected NSCLC; 0.91 (0.76 -1.10) in six studies on any stage NSCLC; 0.57 (0.41 -0.78) in five studies on squamous cell cancer; 0.75 (0.61 -0.93) and 0.71 (0.61 -0.83) respectively for five studies detecting Bcl-2 by immunohistochemistry with Ab clone 100 and for 13 studies assessing Bcl-2 with Ab clone 124; 0.92 (0.73 -1.16) for four studies on small cell lung cancer; 1.26 (0.58 -2.72) for three studies on neuroendocrine tumours. In NSCLC, Bcl-2 expression was associated with a better prognosis. The data on Bcl-2 expression in small cell lung cancer were insufficient to assess its prognostic value.

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Section: Discussionmentioning

confidence: 99%

Role of Bcl-2 as a prognostic factor for survival in lung cancer: a systematic review of the literature with meta-analysis

et al. 2003

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“…Before incubation with the primary antibodies the slides were subjected to antigen retrieval using a boiling solution of 0.01 M citrate buffer pH 6.0 (Cattoretti et al, 1993) in a microwave oven at 700 W and cooled down to room temperature in citrate buffer for 2 h. After washing in PBS, an ovemight incubation followed with mouse monoclonals anti-human ,B-catenin (1:4000, Transduction Laboratories) and anti-E-cadherin, clone HECD-1 (1:500, Zymed). Biotin-labelled rabbit anti-mouse immunoglobulins and a biotinylated HRP-streptavidin complex (both Dako) were subsequently applied for 30 min each with washes in PBS in between.…”

Section: Polymerase Chain Reaction (Pcr) Techniquementioning

confidence: 99%

Loss of heterozygosity for defined regions on chromosomes 3, 11 and 17 in carcinomas of the uterine cervix

Kersemaekers

Hermans²,

Fleuren³

et al. 1998

Br J Cancer

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Summary Loss of heterozygosity (LOH) frequently occurs in squamous cell carcinomas of the uterine cervix and indicates the probable sites of tumour-suppressor genes that play a role in the development of this tumour. To define the localization of these tumour-suppressor genes, we studied loss of heterozygosity in 64 invasive cervical carcinomas (stage IB and IIA) using the polymerase chain reaction with 24 primers for polymorphic repeats of known chromosomal localization. Chromosomes 3, 11, 13, 16 and 17, in particular, were studied. LOH was frequently found on chromosome 11, in particular at 11 q22 (46%) and 11 q23.3 (43%). LOH on chromosome 11p was not frequent. On chromosome 17p1 3.3, a marker (Dl 7S513) distal to p53 showed 38% LOH, whereas p53 itself showed only 20% LOH. On the short arm of chromosome 3, LOH was frequently found (41%) at 3p21.1. The f-catenin gene is located in this chromosomal region. Therefore, expression of f-catenin protein was studied in 39 cases using immunohistochemistry. Staining of 1-catenin at the plasma membrane of tumour cells was present in 38 cases and completely absent in only one case. The tumour-suppressor gene on chromosome 3p21.1 may be ,B-catenin in this one case, but (an)other tumour-suppressor gene(s) must also be present in this region. For the other chromosomes studied, 1 3q (BRCA-2) and 16q (E-cadherin), only sporadic losses (< 15% of cases) were found. Expression of E-cadherin was found in all of 37 cases but in six cases the staining was very weak. No correlation was found between clinical and histological parameters and losses on chromosome 3p, 1 1 q and 1 7p. In addition to LOH, microsatellite instability was found in one tumour for almost all loci and in eight tumours for one to three loci. In conclusion, we have identified three loci with frequent LOH, which may harbour new tumour-suppressor genes, and found microsatellite instability in 14% of cervical carcinomas.

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“…For detection the Elite ABC kit (Vector, Burlingame, CA) was used. The immunohistochemical assay for p16 in para nembedded tissues was performed following a microwave oven antigen retrieval step using hot citrate bu er for 10 min (Cattoretti et al, 1993). Five mm para n sections of oral premalignant lesions were mounted on polysilanecoated slides, dewaxed, rehydrated and incubated with p16 antibody (normal preimmune mouse serum was used as negative control) at a 1 : 400 dilution at 48C overnight.…”

Section: Immunohistochemistrymentioning

confidence: 99%