P53 Scores Research Articles

Evaluation of p53 and Ki67 Expression Profiles in Basal Cell Carcinomas in a Usual and an Unusual Location.

Owing to their importance in cell proliferation in cutaneous malignancies, we aimed to immunohistochemically compare the expression profiles of p53 and Ki67 in basal cell carcinoma (BCC) cases in both a usual and an unusual locations in this study. In this study we included 12 in an unusual location of BCC cases and 21 BCC cases in a usual location. Immunohistochemical expression of p53 and Ki67 antibodies were studied in 33 paraffin-embedded tissue specimens of basal cell carcinoma. We compared the p53 and Ki67 staining scores with clinicopathologic features. The tumor size was found to be greater in BCC cases in an unusual location than those in a usual location. The relationship between age and tumor size was also evaluated in both groups and it was found that tumor size increased with age. A comparative study between the two groups showed no difference p53 and Ki67 expression percentages. There was a linear correlation between the Ki67 and p53 marker staining rates (ρ=0.420; p=0.015). In the samples taken from cases in a usual region, there was a linear and moderate relationship between the markers (ρ=0.513; p=0.017). Median tumor diameter results were similar to the marker staining score (p > 0.05). This is the first study comparing the expression profiles of p53 and Ki67 of BCC cases in an unusual and a usual location. No significant difference was found concerning Ki67 and p53 expression levels between the two groups.

Establishing cut-off points with clinical relevance for bcl-2, cyclin D1, p16, p21, p27, p53, Sox11 and WT1 expression in glioblastoma - a short report.

Glioblastoma (GBM) ranks among the most challenging cancers to treat and there is an urgent need for clinically relevant prognostic and diagnostic biomarkers. Here, we set out to investigate the expression of eight proteins (bcl-2, cyclin D1, p16, p21, p27, p53, Sox11 and WT1) in GBM with the specific aim to establish immunohistochemistry cut-off points with clinical relevance. Immunohistochemistry (IHC) was used to examine protein expression in 55 surgical GBM specimens using H-scores, and IHC cut-off points were established using the Cutoff Finder web platform. Protein co-expression and its correlation with histopathological features were assessed, and cases were classified according to IDH1 mutation status. Survival curves were determined using Kaplan-Meier analyses. Clinical and molecular parameters found to be correlated with overall survival (OS) were tumor size (r= -0.278; p= 0.048), p53 (r= -0.452; p= 0.001), p16 (r= 0.351; p= 0.012) and Sox11 (r= 0.324; p= 0.020). In addition, we found that tumor size correlated with cyclin D1 (r= -0.282; p= 0.037), p53 (r= 0.269; p= 0.041), Sox11 (r= -0.309; p= 0.022) and WT1 (r= -0.372; p= 0.003). Variables found to be significantly associated with IDH1 mutation status were OS (p< 0.01), age (p< 0.01), cyclin D1 (p= 0.046), p16 (p= 0.019) and Sox11 (p= 0.012). Variables found to be significantly associated with a poor survival were tumor size >5cm (p< 0.001), bcl-2 score > 40 (p= 0.034), cyclin D1 score ≤ 70 (p= 0.004), p16 score ≤ 130 (p= 0.005), p53 score > 20 (p= 0.003), Sox11 score ≤ 40 (p< 0.001) and WT1 score ≤ 270 (p= 0.02). Correlations between protein biomarkers and main clinical GBM variables were identified. The establishment of distinct biomarker cut-off points may enable clinicians and pathologists to better weigh their prognostic value.

A cross-sectional study of p53 expression in patients with squamous cell carcinoma cervix: A hospital-based study

Context: Cervical cancer is the second most common cancer in women in India and the fourth most common cancer in women worldwide. The p53 is a tumor suppressor gene. Overexpression of p53 is detected by immunohistochemistry (IHC) and it indicates a worsened prognosis in some malignancies. Aims: The present study was undertaken to assess the expression of p53 by IHC in normal cervical epithelium and squamous cell carcinoma (SCC) cervix. The role of p53 as a potential biomarker by correlating p53 expression with clinico-pathological parameters was also studied. Settings and Design: Cross-sectional study. Subjects and Methods: A total of 50 cases of cervical biopsy or hysterectomy specimens of SCC cervix and fifty cases of hysterectomy specimens for other gynecologic causes were included in the study. IHC was used to study the p53 expression in normal and neoplastic cervical epithelium. The p53 positivity was assessed by using a semi quantitative method in SCC cervix. The p53 expression was correlated with various clinico-pathological parameters. Statistical Analysis Used: Data collected was analyzed using appropriate statistical test. Results: All the fifty cases (100%) of SCC cervix showed positive p53 expression. The normal cervical epithelium in 90% cases was p53 negative and 10% showed basal layer positivity. A statistically significant correlation was seen between p53 score and menopausal status ( P = 0.010). Conclusions: Overexpression of p53 is suggestive of high grade of tumor.

Evaluation of Invasive Squamous Cell Carcinoma, Seborrheic Keratosis and Verruca Vulgaris in Superficial Shave Biopsies using p16, p53, p63, and PHLDA1 Immunohistochemistry

Occasionally, the distinction between malignant and benign is challenging in superficial shave biopsies of squamoproliferative lesions. This phenomenon is compounded by the increasing prevalence of conditions encountered that weaken the immune system, such as chemotherapy, immune deficiency diseases, and antirejection medications for organ transplantation that have all been shown to increase the risk of the development of squamous cell carcinoma that are sampled. We collected 30 cases (10 invasive SCC, 10 SK and 10 VV) and performed immunohistochemical staining using a panel approach composed of markers important for proliferation and cell cycle, including Ki-67, p16, p53, and PHLDA1. The results demonstrate that the invasive SCC group was enriched for high PHLDA1 (80% with PHLDA1 score=3, 100% with PHLDA1 score ≥ 2) and high p53 (50% of SCC with p53 score ≥ 2 vs. 60% of SK and 90% of VV with p53 score=1). The SK group was enriched for low p16 (100% with p16 score ≤ 1) and high p63 scores (100% with p63 score=3). A panel approach may be utilized to help in the distinction between benign keratoses and carcinoma and may be increasingly critical to promote quality care.

Expression of Phosphatase and Tensin Homologue, phospho-Akt, and p53 in Acral Benign and Malignant Melanocytic Neoplasms (Benign Nevi, Dysplastic Nevi, and Acral Melanomas).

BackgroundThe role of the phosphatidylinositol-3 kinase signaling pathway in the development of acral melanoma has recently gained evidence. Phosphatase and tensin homologue (PTEN), one of the key molecules in the pathway, acts as a tumor suppressor through either an Akt-dependent or Akt-independent pathway. Akt accelerates degradation of p53.ObjectiveWe assessed the expression of PTEN, phospho-Akt (p-Akt), and p53 by immunohistochemistry in benign acral nevi, acral dysplastic nevi, and acral melanomas in the radial growth phase and with a vertical growth component.MethodsTen specimens in each group were included. Paraffin-embedded specimens were immunostained with antibodies for PTEN, p-Akt, and p53. We scored both the staining intensity and the proportion of positive cells. The final score was calculated by multiplying the intensity score by the proportion score.ResultsAll specimens of benign acral nevi except one showed some degree of PTEN-negative cells. The numbers of p-Akt and p53-positive cells were higher in acral dysplastic nevi and melanoma than in benign nevi. P-Akt scores were 1.7, 1.8, 2.6, and 4.4, and p53 scores were 2.0, 2.1, 3.8, and 4.1 in each group. PTEN and p-Akt scores in advanced acral melanoma were higher than in the other neoplasms.ConclusionThe expression of PTEN was decreased and the expression of p-Akt was increased in acral melanoma, especially in advanced cases. The PTEN-induced pathway appears to affect the late stage of melanomagenesis. Altered expression of p-Akt is thought to be due to secondary changes following the loss of PTEN.

Expression level of p53 in Tumor Tissue of Patients with Acute Leukemia and Its Clinical Significance

To study the expression level of p53 in tumor tissue of patients with acute leukemia (AL) and its clinical significance. From April 2013 to April 2015, 80 patients with AL in our hospital were chosen as leukemia group, at the same time, 50 patients with non-hematologic diseases were chosen as control group. All the patients were detected by bone marrow smear mean and p53 staining. The positive rate of p53 and staining score were compared between 2 groups. The clinical data of leukemia group were collected, and the correlation of clinical features with p53 expression was analyzed. In the control group, the positive rate of p53 was 6.00%, the staining score was (0.2 ± 0.1); in the leukemia group, the positive rate of p53 was 73.75%, the staining score was (1.9 ± 0.4), the difference was statistically significant (P < 0.05). The expression of p53 significantly correlated with the blood routine indicators, clinical manifestation, multiple infiltration and curative effects (P < 0.05), and the p53 expression not correlated statistically with the sex, age, anamnesis, family history (P > 0.05). Compared with the patients with non-hematologic diseases, the expression level of p53 in the AL patients increases significantly, the p53 expression correlates significantly with the blood routine indicators, clinical manifestation and curative effect of the patients.

Abstract C32: PTEN and p53 expression in endometrial cancer correlated with clinicopathological phenotype.

Abstract p53 overexpression and loss of PTEN expression have been implicated in the pathogenesis of endometrial cancer. Our aim was to correlate expression levels of these proteins with clinicopathological parameters to characterise their utility as biomarkers. Immunohistochemistry was done on 49 cases of formalin fixed paraffin embedded endometrial cancer tissue for p53 and 47 for PTEN expression. For p53 staining the Hue Saturation and Intensity mathematical model was used to evaluate different degrees of positivity of the immunohistochemical stain. The quick score was used for p53 and immunoreactive score for PTEN was done to score immunoreactivity. For PTEN, average percentage of positive cells and intensity was evaluated and the immunoreactive score was calculated. Both these scoring methods incorporated percentage of positive cells and staining intensity. Univariate Cox regression analysis for cause specific survival showed a hazard ratio of 9.2 for p53 overexpression (P&amp;lt;0.05). High levels of P53 were significantly associated with histologic type, grade, LVI and recurrent disease. There was no significant association between PTEN expression and clinicopathological features but it was noted that the 8 patients who relapsed were PTEN negative. Cause specific survival could not be assessed for PTEN expression due to the lack of events in this group. PTEN and p53 (n=46) combined analysis failed to demonstrate a significant association on prognostic features in this group. Overexpression of P53 has a negative effect on disease specific survival and was associated with poor prognostic features. We did not demonstrate a significant association with clinicopathological features assessed and PTEN expression. However of the 8 patients who relapsed all were PTEN negative and 7 were p53 positive. Dual assessment of p53 and PTEN warrants further investigation as a prognostic indicator. Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C32. Citation Format: Bihani Kularatne, Arrigo Capitanio, Rupali Arora, Philippa Jones, Andre Lopes, Jennifer Paterson, Rebecca Kristeleit. PTEN and p53 expression in endometrial cancer correlated with clinicopathological phenotype. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C32.

Precursor Lesions and Prognostic Factors in Primary Peritoneal Serous Carcinoma

Primary peritoneal serous carcinoma (PPSC) is uncommon and precursor lesions and prognostic factors are incompletely understood or described. Charts of 22 women with PPSC were reviewed for clinical and pathology data. Glass slides were reviewed for areas of PPSC, ovarian surface epithelium (OSE), ovarian cortical inclusion cysts (OCICs), tubal epithelial atypia (TEA), and serous tubal intraepithelial carcinoma (STIC). p53 and p16 immunohistochemical staining was scored and expression between the sites was compared. PPSC outcome was correlated with biomarker expression and clinicopathologic variables. p53 and p16 scores were significantly higher among PPSC and tubes than OSE and OCICs. Approximately 64% of PPSC, 46% of fallopian tubes, and none of the OSE overexpressed both biomarkers. OCICs were more frequently negative. Tubal pathologies of STIC, TEA and/or p53 signatures were present in approximately 46% of PPSC cases and the association with tubal p53 overexpression was significant. Statistically significant associations between tubal pathologies, OSE and OCICs biomarker expression profiles and their PPSC expression profiles did not occur. The median overall survival was 53 months and low-grade tumors had a better prognosis (P=0.02). A role for p53 and p16 overexpression in the formation of some PPSC, and p53 overexpression in the generation of precursor tubal pathology was identified. The high frequency of tubal pathology overexpressing p53 and low frequency of OSE and OCICs expressing either biomarker highlights a possible precursor role for the tube in some PPSC. Tumor grade was the only significant prognostic factor.

A study of histopathological spectrum and expression of Ki-67, TP53 in primary brain tumors of pediatric age group

Objectives:The primary brain tumors are the second most common cause of death due to malignancies in children. This study was done to analyze the histological spectrum of primary brain tumors in children and also to find out the expression of p53 and Ki67 in some of the common pediatric brain tumors.Materials and Methods:This study was done over a period of 2.5 years. The patients were followed up until 6 months to determine the outcome. We examined H and E sections from 61 pediatric brain tumors and also performed immunohistochemical stains with p53 and Ki67 on 52 of these samples.Results:Of the 61 cases of pediatric brain tumors the commonest were pilocytic astrocytomas and medulloblastomas both constituting 22.9% of total cases, followed by high grade gliomas, that is, anaplastic astrocytoma and glioblastoma taken together (14.7%), diffuse astrocytomas (11.4%), ependymomas (8.1%), and oligodendrogliomas (4.9%). Other cases comprised craniopharyngiomas, astroblastomas, and gangliocytoma. The mean age of presentation was 9.3 years, male children being more commonly affected. Ki67 labeling index (LI) and p53 expression in pilocytic astrocytomas and diffuse astrocytomas were significantly lower than that of high-grade astrocytomas. However, there was no significant difference of expression of these two antigens in pilocytic astrocytomas and diffuse astrocytomas. It was found that Ki67 LI was a better marker for distinguishing between grades of astrocytoma than p53 (P=0.000 and P=0.002, respectively). The survival in cases of pilocytic astrocytomas was far better than high-grade gliomas. However, there was no significant difference in survival between pilocytic astrocytoma and diffuse infiltrating astrocytoma. There was significant positive correlation between expression of p53 and Ki67 LI in cases of medulloblastomas. Both p53 (P=0.002) and Ki67 LI (P=0.000) taken individually correlated well with survival in these cases. Also, Ki67 LI is better predictor of outcome than p53.Conclusion:From this study, it can be concluded that Ki67 and p53 score correlated well with the grade of astrocytoma; however, Ki67 is a better marker for differentiating between the grades of astrocytoma than p53. Also, Ki67 LI is a better prognostic factor than p53 in case of medulloblastomas.

Increased fibroblast density in actinic cheilitis: association with tryptase‐positive mast cells, actinic elastosis and epithelial p53 and COX‐2 expression

Actinic cheilitis (AC) is characterized by epithelial and connective tissue alterations caused by ultraviolet sunlight overexposure known as photodamage. Fibroblasts have been linked to photodamage and tumor progression during skin carcinogenesis; however, their role in early lip carcinogenesis remains unknown. The aim of this study was to assess the density of fibroblasts in AC and normal lip (NL) samples and determine their association with markers of lip photodamage. Fibroblasts, mast cells, p53, COX-2, and elastin were detected in NL (n = 20) and AC (n = 28) biopsies using immunohistochemistry/histochemistry. Mast cell and fibroblast density and epithelial p53 and COX-2 expression scores were then obtained. Elastosis was scored 1-4 according to elastin fiber density and tortuosity. Fibroblasts, mast cells, p53, COX-2, and elastosis were increased in AC as compared to NL (P < 0.001). Multivariate analysis showed an association between fibroblast and mast cell density at the papillary and reticular areas of AC and NL (P < 0.05). Papillary fibroblast density was also associated with epithelial p53 and COX-2 expression (P < 0.05). Increased fibroblast density, both papillary and reticular, was found in the high elastosis group (scores 3-4) as compared to the low elastosis group (scores 1-2) (P < 0.01). Increased reticular mast cell density was detected only in the high elastosis group (P < 0.01). Fibroblasts are increased in AC, and they are associated with mast cell density, epithelial p53 and COX-2 expression, and actinic elastosis. Therefore, fibroblasts may contribute to lip photodamage and could be considered useful markers of early lip carcinogenesis.

Precursors of Endometrial Clear Cell Carcinoma

The recognition of morphologically identifiable lesions which may confer an increased risk for subsequent development of an invasive malignancy offers an opportunity to investigate and better understand the molecular-genetic etiopathogenesis of the well-developed tumor, and potentially, to administer a therapeutic intervention before its development. In contrast to uterine endometrioid and serous carcinomas, very little is known about the potential precursor lesions of endometrial clear cell carcinoma (ECCC). In our routine practice, we have noted the presence of a spectrum of atypical glandular changes in the endometria adjacent to ECCC or endometrial carcinomas with a clear cell component, which on the basis of current criteria, would not qualify for any specific designation. We hypothesize that these lesions represent the earliest morphologically recognizable precursor lesions of ECCC and systematically characterize their clinicopathologic features herein. Thirty archived cases of pure ECCC (n=14) or mixed endometrial carcinomas with a >10% clear cell component (n=16) were retrieved and the "normal" endometria adjacent to the malignancies were evaluated in detail. Thirty-eight benign uteri and 30 uteri with classic endometrial endometrioid carcinoma (EEC) served as controls. All cases were reviewed in a blinded fashion. Putative precursor lesions (PPL) were searched for and identified microscopically. The lesions were typically isolated glands or surface epithelium (within an otherwise normal endometrial region) that displayed cytoplasmic clarity and/or eosinophilia with varying degrees of nuclear atypia. Twenty-seven (90%) of the 30 cases had at least 1 PPL. In contrast, PPL were identified neither in the benign uteri nor in endometrioid carcinoma control groups (P<0.001). A total of 67 foci of PPL were identified in the 27 cases with an average of 2.5 foci per case. The immunohistochemical expression of p53, mib-1, estrogen receptor (ERs), and progesterone receptor in the benign endometria, ECCC, and the PPL were evaluated on all 27 cases. The mean p53 scores for the benign endometria, PPL, and ECCC were 0, 4.5, and 6.2, respectively. Parallel values for mib-1 were 15%, 45%, and 63%. ER/progesterone receptor indices for benign endometria, PPL, and carcinoma were 90/80, 21.52/4.61, and 11/4, respectively. The PPL described herein have a morphologic and immunophenotypic profile which seems to be distinct from both the benign endometria in which they reside and the adjacent areas of ECCC. The high frequency of association of these lesions with ECCC, their frequent occurrence as isolated lesions within otherwise "benign-appearing" endometria, and their continuous spectrum of nuclear atypia from minimum (grade 1, cytologic atypia falls short of ECCC cells) to maximum (grade 3, cytologically identical to ECCC cells), argues in favor of our hypothesis that these may represent precursor lesions of ECCC. Further studies are required to conclusively define the nature of these lesions. However, such studies can only be performed if diagnostic surgical pathologists recognize, highlight, and segregate these lesions for further analysis.

P53 and erbB-2 are not associated in matched cases of primary and metastatic ovarian carcinomas.

Ovarian cancer has a high mortality rate largely due to the limited number of ovarian carcinomas detected at an early stage. Understanding the molecular changes occurring during the progression of ovarian carcinoma would aid in the development of therapies that may inhibit or target metastasis. Primary and metastatic lesions from 54 and 40 patients with advanced ovarian carcinoma, respectively (including matched primary and metastatic lesions from 30 patients) were evaluated for nuclear accumulation of p53 (clone BP53-12-1) and cytoplasmic and membranous immunostaining of p185 erbB-2 (clone 3B5) by immunohistochemistry. No differences in the immunostaining of p53 and p185erbB-2 (cytoplasm or membrane) were observed between primary and metastatic lesions of the matched cases. Similarly, no differences in the proportion of positive cases of p53 between primary and metastatic lesions of the matched cases was observed. Thus, novel therapies that target p53 or p185erbB-2 can utilize specimens from either primary or metastatic lesions to characterize these targets prior to therapy. Spearman correlations between p53 and p185erbB-2 (cytoplasm or membrane) immunohistochemistry scores were insignificant for the matched cases, all primary lesions, and all metastatic lesions. Also, no significant associations occurred between nuclear accumulation of p53 (positive versus negative) and phenotypic expression of p185erbB-2 (cytoplasm or membrane) immunostaining scores for the matched cases, all primary lesions, and all metastatic lesions. Thus, the nuclear accumulation of p53 and immunostaining of p185erbB-2 in the cytoplasm or on the cellular membranes are independent.

EGFR overexpression and radiation response in glioblastoma multiforme

Purpose: Recent studies have suggested relative radioresistance in glioblastoma multiforme (GM) tumors in older patients, consistent with their shorter survival. Two common molecular genetic abnormalities in GM are age related: epidermal growth factor receptor (EGFR) overexpression in older patients and p53 mutations in younger patients. We tested whether these abnormalities correlated with clinical heterogeneity in GM response to radiation treatment.Methods and Materials: Radiographically assessed radiation response (5-level scale) was correlated with EGFR immunoreactivity, p53 immunoreactivity, and p53 exon 5–8 mutation status in 170 GM patients treated using 2 prospective clinical protocols. Spearman rank correlation and proportional-odds ordinal regression were used for univariate and multivariate analysis.Results: Positive EGFR immunoreactivity predicted poor radiographically assessed radiation response (p = 0.046). Thirty-three percent of tumors with no EGFR immunoreactivity had good radiation responses (>50% reduction in tumor size by CT or MRI), compared to 18% of tumors with intermediate EGFR staining and 9% of tumors with strong staining. There was no significant relationship between p53 immunoreactivity or mutation status and radiation response. Significant relationships were noted between EGFR score and older age and between p53 score or mutation status and younger age.Conclusion: The observed relative radioresistance of some GMs is associated with overexpression of EGFR.

STAGE PROGRESSION IN TA PAPILLARY UROTHELIAL TUMORS: RELATIONSHIP TO GRADE, IMMUNOHISTOCHEMICAL EXPRESSION OF TUMOR MARKERS, MITOTIC FREQUENCY AND DNA PLOIDY

We studied 363 patients with stage Ta bladder tumors during long-term followup who were classified according to the 1998 WHO and International Society of Urological Pathology consensus classifications. We determine whether various immunohistochemical and molecular markers could predict tumor progression. A total of 680 patients in western Sweden with a first diagnosis of bladder carcinoma in 1987 and 1988 were registered and followed for at least 5 years. There were 363 (53%) tumors that were papillary stage pTa. The tumors were classified as papillary urothelial neoplasm of low malignant potential in 95 patients, low grade papillary urothelial carcinoma in 160 and high grade carcinoma in 108. Of the patients in the latter group 95 were subdivided into WHO grade 2 and 13 into WHO grade 3. Tissue from the primary tumors that progressed in stage during followup was further analyzed with immunohistochemical methods (p21, p53, Ki67 and pRb), DNA ploidy and mitotic frequency. The results were compared with those in matched controls (nonprogressors). Recurrence developed in 35% of patients with papillary urothelial neoplasm of low malignant potential compared to 71% with low grade urothelial carcinoma and 73% with high grade carcinoma (p <0.0001). No papillary urothelial neoplasm of low malignant potential progressed in stage. Disease progressed in 4% of patients with low grade compared to 23% with high grade carcinoma (p <0.0001). Of the patients with WHO grade 3 disease progressed in 45% compared to grade 2 in 20% (p <0.0011). At first diagnosis p53 score was significantly higher (p <0.0022) among patients with WHO grade 2 carcinoma which later progressed compared to that in matched controls but there was no significant difference regarding the other markers. In contrast to grade 2 most grade 3 carcinoma was aneuploid, had high mitosis frequency, high p53 and Ki67 scores as well as loss of retinoblastoma gene expression. The 1988 WHO and International Society of Urological Pathology consensus classifications divide noninvasive papillary bladder tumors into 3 subgroups with different clinical behavior, which seems to be an advantage compared with the 1973 WHO classification. A disadvantage is that the high grade carcinoma group contains 2 subgroups with different progression rates and immunohistochemical marker profiles, corresponding to the 1999 WHO grades 2 and 3. Grade 2 tumors in patients that progressed in stage years later seem to have different immunohistochemical and molecular marker profiles compared to those in matched controls.

Immunohistochemically stained markers (p53, PCNA, bcl-2) in dysplastic lesions of the larynx

The percentage of malignant transformation of laryngeal dysplastic lesions is difficult to estimate. There is a need for new histological markers which could enable more objective assessment of the premalignant stages of the larynx and help in estimation of the potential of future neoplastic progression. We performed a retrospective study to determine whether immunohistochemical staining for the proliferating cell nuclear antigen (PCNA), tumour suppressor gene protein p53 and anti-apoptotic protein bcl-2 may be prognostic factors in laryngeal epithelial lesions. Staining was performed on 57 paraffin-embedded biopsies from patients with clinically detected precancerous stages of the larynx. Histopathologic examination revealed normal epithelium in six cases, mild dysplasia in 20 cases, moderate dysplasia in 18 cases, severe dysplasia in seven cases, CA in situ in four cases, papilloma in one case and CA invasivum in one case. The p53 count in mild and moderate dysplasia was 26.8 and 38.6%, respectively. This difference was statistically significant. There was significant correlation between PCNA and p53 scores. There was also a relationship between the scores of these markers and bcl-2 expression. In ten out of 45 cases of dysplastic lesions the invasive cancer developed in 4 years of follow-up. The correlation between PCNA score and malignant progression of the dysplastic lesions was on the statistical borderline. There was significant relationship between malignant transformation and age of the patients.

Sun-exposure- and aging-dependent p53 protein accumulation results in growth advantage for tumour cells in carcinogenesis of nonmelanocytic skin cancer.

Three hundred and sixteen patients with nonmelanocytic skin cancer, including 46 cases of Bowen's disease (BOD), 134 cases of squamous cell carcinoma (SCC), and 136 cases of basal cell carcinoma (BCC), were examined immunohistochemically using monoclonal antibody DO-7 to assess p53 protein accumulation related to sun exposure and ageing, and growth and differentiation of skin cancer and its precursors. The rates of p53 immunostaining of BOD, SCC and BCC were 80.4%, 76.1% and 70.6%, respectively. p53-positive cells were present not only in cancer nests, but also in dysplastic and even morphologically normal epidermis adjoining cancers. Sun exposure was statistically correlated with the p53 immunostaining scores in morphologically normal epidermis of the three skin cancers and in cancer nests of SCC and BCC. The positivity and score of p53 protein often differed significantly among the three types of cancer, especially in regions of dysplasia. Interestingly, differentiation of SCC was correlated with individual p53 scores for dysplasia and cancer nests, especially for dysplasia. BOD, as the precursor of SCC, demonstrated the strongest p53 expression. Furthermore, 12.3% cases with p53 negative cancer nests showed p53-positive reaction in dysplasia and in morphologically normal epidermis. It seems that the accumulation of p53 protein plays a part in precancerous lesions and in the genesis of more highly differentiated types of skin cancer and affects mainly the growth of tumour cells rather than their differentiation. For BCC, however, age was significantly related to p53 expression. Our findings suggest that overexpression of p53 in normal skin and cancer nests of SCC and BCC is significantly related to sun exposure, that the expression of p53 in BCC is an age-dependent process, and that the early accumulation of p53 protein may be a useful predictor for the detection of nonmelanocytic skin cancer.

Association of Bcl-2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis

Bcl-2 protein is believed to play a role in neoplasia by inhibiting tumor cell apoptosis. To assess its contribution to gallbladder tumorigenesis and cancer progression, an immunohistochemical study was performed. Fifteen adenomas and 68 adenocarcinomas were immunohistochemically and histopathologically investigated for the relation of Bcl-2 expression to p53 status, apoptosis (apoptotic index, AI), and proliferation activity (mitotic index, MI; Ki-67 labeling index, Ki-67 LI). The Bcl-2 score, based on intensity and extent, decreased in the order of adenoma, well-differentiated, and moderately to poorly differentiated adenocarcinoma. Early stage carcinomas demonstrated significantly higher Bcl-2 scores than their advanced counterparts (P < 0.05). On the other hand, p53 score, MI, Ki-67 LI, and AI increased in the same order. The Bcl-2 negative adenocarcinomas displayed higher AI and AI-to-MI ratios than the Bcl-2 positive group, especially in the early stage, well-differentiated lesions. A significantly positive correlation between MI(r=0.549) or Ki-67 LI(r = 0.446) and AI was observed. In early stage carcinomas, adenomatous components in the lesions were found more frequently in the polypoid lesions than in the nonpolypoid lesions (P < 0.05). Expression of Bcl-2 protein in gallbladder tumors appears to be positively associated with tumor cell differentiation and inversely with tumor progression. It may thus play a role in regulating carcinoma growth, especially in the early stage of tumorigenesis. It is believed that the polypoid carcinomas may arise from preexisting adenomas but the nonpolypoid carcinomas may arise as de novo carcinoma.

Association of Bcl‐2 protein expression with gallbladder carcinoma differentiation and progression and its relation to apoptosis

BACKGROUND Bcl-2 protein is believed to play a role in neoplasia by inhibiting tumor cell apoptosis. To assess its contribution to gallbladder tumorigenesis and cancer progression, an immunohistochemical study was performed. METHODS Fifteen adenomas and 68 adenocarcinomas were immunohistochemically and histopathologically investigated for the relation of Bcl-2 expression to p53 status, apoptosis (apoptotic index, AI), and proliferation activity (mitotic index, MI; Ki-67 labeling index, Ki-67 LI). RESULTS The Bcl-2 score, based on intensity and extent, decreased in the order of adenoma, well-differentiated, and moderately to poorly differentiated adenocarcinoma. Early stage carcinomas demonstrated significantly higher Bcl-2 scores than their advanced counterparts (P < 0.05). On the other hand, p53 score, MI, Ki-67 LI, and AI increased in the same order. The Bcl-2 negative adenocarcinomas displayed higher AI and AI-to-MI ratios than the Bcl-2 positive group, especially in the early stage, well-differentiated lesions. A significantly positive correlation between MI(r = 0.549) or Ki-67 LI(r = 0.446) and AI was observed. In early stage carcinomas, adenomatous components in the lesions were found more frequently in the polypoid lesions than in the nonpolypoid lesions (P < 0.05). CONCLUSIONS Expression of Bcl-2 protein in gallbladder tumors appears to be positively associated with tumor cell differentiation and inversely with tumor progression. It may thus play a role in regulating carcinoma growth, especially in the early stage of tumorigenesis. It is believed that the polypoid carcinomas may arise from preexisting adenomas but the nonpolypoid carcinomas may arise as de novo carcinoma. Cancer 1999;85:318–25. © 1999 American Cancer Society.

The prognostic value of p53 nuclear overexpression and MIB-1 as a proliferative marker in transitional cell carcinoma of the bladder

There is controversy regarding the value of biologic markers as prognostic indicators independent of clinicopathologic parameters in transitional cell carcinoma (TCC) of the bladder. The authors examined the prognostic value of p53 tumor suppressor gene expression and the proliferative marker MIB-1 in TCC of the bladder. Fresh frozen samples from 114 TCCs of the bladder and 13 normal bladders were studied by immunohistochemistry, using monoclonal antibodies MIB-1 for proliferation and PAb 1801 for p53 nuclear overexpression. Scores were determined in each case by counting at least 500 nuclei per slide in 3-5 selected regions. Patients were stratified for both markers into two groups for time-event analysis, according to the median number of nuclei stained. Patients with nuclear staining below the median value of the score were considered negative in the statistical analysis. Quantitative immunostaining was analyzed in relation to the time to recurrence, progression, and cancer death, and compared with clinical and pathologic parameters for prognostic significance in univariate and multivariate analysis (stepwise logistic regression). Median nuclear overexpression of p53 was 22% and that of MIB-1 28%. There was a strong association between proliferation and p53 nuclear overexpression (P < 0.0001). Progression free and disease specific survival rate estimates (log rank test) were significantly lower in patients with p53 or MIB-1 scores above 22% and 28%, respectively. Multivariate analysis indicated that stage as well as p53 and MIB-1 immunostaining provided independent prognostic information. Quantitative immunohistochemical evaluation of nuclear p53 and MIB-1 immunostaining inexpensively provides prognostic indicators in cases of TCC of the bladder.

P53, bcl-2 and Retinoblastoma Proteins as Long-Term Prognostic Markers in Localized Carcinoma of the Prostate

Purpose The accumulation of p53 and bcl-2 gene products as well as the loss of the retinoblastoma (Rb) gene product have been associated with prostate cancer progression. We assessed whether the levels of immunoreactivity for p53, Rb and bcl-2 are better long-term predictors of disease specific survival than conventional pathological parameters of the primary tumor, such as Gleason score, capsular penetration, seminal vesicle invasion, and percent tumor in the specimen, in patients with clinically localized prostate cancer treated with radical prostatectomy. Materials and Methods A total of 71 patients with clinical stages A1 to B2 adenocarcinoma of the prostate underwent radical prostatectomy after a negative metastatic evaluation. No neoadjuvant or adjuvant treatments were given and causes of death were recorded. Prostatectomy specimens were analyzed to determine the conventional pathological parameters, and p53, Rb and bcl-2 immunohistochemical staining. Univariate and multivariate analyses were done to determine the independent contributions of p53, Rb and bcl-2 in predicting survival. Results On multivariate analysis the independent factors predicting disease specific survival were p53 staining score (p <0.001) and Rb staining score (p <0.001). In patients with p53 immunoreactive tumors the 15-year disease specific survival was 38% compared to 87% for those with less immunoreactivity. Analysis of Rb immunoreactivity for 15-year disease specific survival yielded 92 and 66% high and low staining levels, respectively. Best subset analysis revealed that the combination of p53 score and Rb score yielded the best predictive value for disease specific survival. Conclusions p53 and Rb immunohistochemical staining scores were independent predictors of disease specific survival and were superior to conventional pathological prognostic factors of the primary tumor. These findings lay the groundwork for the prospective study of these markers in patients treated with radical prostatectomy.