p53 immunohistochemistry as an independent prognostic factor for superficial transitional cell carcinoma of the bladder.

Although patients with superficial bladder cancer (Ta, T1) have a generally good prognosis, those patients who develop muscle-invasive tumours or metastatic disease at recurrence do poorly clinically. In the current study 69 patients undergoing complete transurethral resection for superficial transitional cell cancer of the bladder were investigated for different clinical and biological characteristics as possible prognostic factors: age, sex, performance of instillation therapy and immunohistochemical determination of mutational inactivation of p53 tumour-suppressor gene (monoclonal antibody PAb 1801) as well as immunohistochemical determination of the proliferation rate by staining for PCNA (proliferating cell nuclear antigen) (monoclonal antibody PC 10). After a median follow-up of 45.8 months, 12 of 14 patients (85.7%) with more than 20% of cells positive for p53 had disease progression with muscle-invasive growth compared with only one of 55 patients (1.8%) negative for p53 (P < 0.01, chi 2 test). During univariate analysis histological grade (G1 vs G2) (P = 0.0373), positivity for PCNA (> 60% of cells) (P = 0.0033) and positivity for p53 (P < 0.001) were significant prognostic factors for disease progression (log-rank test), while during multivariate analysis only positivity for p53 was a significant predictor for relapse of bladder cancer (P = 0.0029) (multivariate Cox regression analysis). The immunohistochemical detection of mutations of the p53 gene has been demonstrated to be a reliable, easily performed and thereby widely available technique for the investigation of fresh-frozen or paraffin-embedded tumour specimens. The results demonstrate the important role of the p53 tumour-suppressor gene protein in the development and for the progression of bladder cancer. If the high prognostic value of p53 mutations in superficial bladder cancer is confirmed in larger prospective trials, more aggressive therapeutic strategies could be discussed for patients with p53 mutations in their tumour specimens.     

Proliferating cell nuclear antigen and p53 expression as prognostic factors in T1-2MO prostatic adenocarcinoma

The expression of proliferating cell nuclear antigen and p53 protein was analysed by immunocytochemical methods (PC10, CMI antisera) in 139 patients with T1-2MO prostatic adenocarcinomas followed-up for > 12 years. p53 protein was expressed in 21 (15%) tumours (15%), the fraction of positive nuclei being very low (mean SE, 1% ± 0.7%). Accumulation of p53 protein in epithelial cells was independent of tumour stage and Gleason score, and had no effect on prognosis. In 4 cases, p53 protein was expressed only in stromal cells. The fraction of PCNA-positive nuclei (evaluable in 116 cases) was higher in T2 than in T1 tumours (p < 0.001); furthermore, high Gleason score was positively correlated with PCNA positivity (p < 0.001). A finding of over 5% of PCNA-positive nuclei predicted progression in T and M categories and were a sign of poor outcome. The fraction of PCNA-positive stromal-cell nuclei was related to T-category with a borderline significance (p = 0.06). In a multi-variate analysis of the prognostic factors, independent predictors of survival included Gleason score (p < 0.001), fraction of PCNA-positive nuclei (p = 0.013), observation before therapy (p = 0.05), and T-category (p = 0.07) in that order of significance. The results suggest that overexpression of p53 protein is of marginal prognostic value in local prostatic adenocarcinomas, whereas direct measurement of cell proliferation by PCNA immunolabelling provides important prognostic information in T1-2MO tumours, in addition to the Gleason score.     

p53、c-erbB2基因、增殖细胞核抗原表达与卵巢上皮性癌预后的关系

背景与目的：p53、c-erbB2基因和增殖细胞核抗原（proliferating cell nuclear antigen,PCNA)的表达与卵巢癌预后关系问题已有报道,由于研究方法、样本大小等方面的不同,结果存在差异。为进一步探讨p53、c-erbB2基因和PCNA表达与卵巢上皮性癌预后的关系,我们进行了本研究。材料与方法：1990年3月1日至1994年3月31日间确诊的卵巢上皮性癌共84例,采用标记的链白素－生物素辣根过氧化酶复合物（LSAB）免疫组化方法检测癌组织中p53、c-erbB2基因和PCNA的表达,使用SPSS8．0版分析它们的表达与卵巢癌患者平均生存期、5年生存率之间的关系。结果：本组84例癌组织中,p53、c-erbB2和PCNA表达的阳性率分别为58．3％（49／84）、77．5％（65／84）和100％（84／84）。单因素分析显示,c-erbB2过度表达和PCNA表达强度与患者的平均生存期、5年生存率均呈负相关（P值分别为0．05和小于0．01）,未发现p53蛋白表达与患者的平均生存期、5年生存率之间存在相关性（P＞0．05）。多因素分析发现卵巢上皮性癌的预后与临床分期、组织分化程度有关,而与p53、c-erbB2和PCNA的表达均无相关性。结论：p53的表达与卵巢上皮性癌的预后无关,而c-erbB2、PCNA表达对卵巢上皮性癌预后的影响是间接的。临床分期、组织分化程度仍是卵巢上皮性癌独立的预后因素。     

Can p53 nuclear over-expression, Bcl-2 accumulation and PCNA status be of prognostic significance in high-risk superficial and invasive bladder tumours?

AIMS: To evaluate p53 and Bcl-2 expression and proliferating status (PCNA) in subgroups of patients with high-risk superficial and invasive bladder cancer, with relation to cancer progression and death, and to correlate the results with established clinical prognostic factors. METHODS: Paraffin-embedded sections from 42 high-risk superficial (T1G2,T1G3) and 33 invasive (T2-T4aG3 N0M0) tumours were investigated immunohistochemically for p53, Bcl-2 and PCNA. The median follow-up was 52 months. RESULTS: In the cohort of superficial tumours, statistical analysis showed that p53 and PCNA positivity were significant prognostic factors (P-values: 0.008 and 0.006, respectively) for disease-specific death (DSD). When life expectancy was evaluated (log-rank test), p53(+) (P = 0.015) and PCNA(+) (P = 0.017) offered the most accurate prognosis compared to grade, tumour size and multiplicity. Bcl-2 status had no significant effect on patient survival. In the subset of muscle-invasive tumours we failed to demonstrate any important role of p53, Bcl-2 or PCNA positivity. CONCLUSIONS: p53 and PCNA over-expression may offer valuable additional prognostic information in high-risk subgroups of superficial bladder tumours. From our results, Bcl-2 does not appear to contribute significantly to the prognosis of these patients. None of the studied markers offered prognostic information in muscle-invading disease.     

膀胱移行细胞癌中p53突变、bcl-2和PCNA表达的临床意义(英文)

目的探讨膀胱癌中 bcl-2、p53、PCNA 表达与细胞增殖、凋亡和临床病理学参数之间的关系。方法 SABC 免疫组化分折62例(T1G1-G339例,T2-T4aG3 NOM023例)甲醛固定和石蜡包埋的膀胱癌标本 bcl-2、p53和 PCNA 蛋白的免疫反应性。平均随访37个月,24例复发。增殖指数(PI)表示肿瘤细胞中 PCNA 阳性细胞百分比。TUNEL 法检测细胞凋亡,凋亡指数(AI)表示肿瘤细胞中凋亡细胞的百分比。结果 62例膀胱癌中,50例(80.0%)发生 p53突变,与 G1(72.7%)和 G2(78.5%)相比较 G3(91.3%)更多见(P<0.05);pT2期(95.7%)p53突变率较 pTa-1期(74.3%)高(P<0.01)。14例(22.5%)发现有 bcl-2表达,bcl-2表达阳性率 G3明显高于 G1和 G2(P<0.05),与分期无关(P>0.05)。Bcl-2表达与 p53突变无关。在膀胱癌中,PI 为17.2%～41.8%(平均为22.4%),AI 为1.9%～3.5%(平均为2.9%)。统计分析显示 PI 与肿瘤分级、分期关系密切,AI 与肿瘤的分级有明显关系。结论结果表明,p53突变与浸润性行为呈正相关。在膀胱癌中 p53和 PCNA 过表达可能能提供有价值的预后信息。随着肿瘤的进展,肿瘤细胞过度增殖可能伴有频繁的凋亡,但增殖指数的增加明显强于凋亡指数的增加。     

Pre-treatment lymphocytopaenia is an adverse prognostic biomarker in muscle-invasive and advanced bladder cancer

BackgroundPre-treatment lymphocytopaenia may result from cytokines secreted by the tumour microenvironment in association with aggressive tumour biology. We sought to establish the prognostic significance of lymphocytopaenia in muscle-invasive and advanced bladder cancer.Patients and methodsSeventy-four patients with muscle-invasive bladder cancer treated with radical chemoradiotherapy and 131 patients with advanced bladder cancer treated with palliative chemotherapy were included in the study. The absolute lymphocyte count on the first day of treatment was recorded. Invasive local or systemic recurrence in the muscle-invasive bladder cancer cohort and all-cause mortality in the advanced bladder cancer cohort were defined as survival end points. Receiver operating characteristic (ROC) curve analysis was utilized to determine the cut-off for defining lymphocytopaenia in the muscle-invasive bladder cancer cohort followed by multivariable analysis in a model evaluating the following variables: anaemia, neutrophilia, tumour stage, hydronephrosis and neoadjuvant chemotherapy. Subsequently, lymphocytopaenia was assessed in a multivariable model of the advanced bladder cancer cohort analysing the following prognostic variables: neutrophilia, anaemia, performance status and presence of bone or visceral metastases. A further analysis was carried out evaluating absolute lymphocyte count as a continuous variable.ResultsAn absolute lymphocyte count of 1.5 × 10⁹/l was determined as the cut-off on ROC curve analysis in the muscle-invasive bladder cancer cohort, and multivariate analysis revealed that only lymphocytopaenia was predictive for inferior outcome in this cohort. In the advanced bladder cancer cohort, lymphocytopaenia [hazard ratio (HR) 1.6, 95% confidence interval (CI) 1.1–2.4; P = 0.02] and performance status (HR 1.7, 95% CI 1.0–2.7; P = 0.047) were adverse prognostic factors in the binary variable multivariate model. Absolute lymphocyte count was the sole significant factor when analysed as a continuous variable (HR 0.66, 95% CI 0.5–0.87; P = 0.003).ConclusionPre-treatment lymphocytopaenia is an independent adverse prognostic factor in both muscle-invasive and advanced bladder cancer. It may be a manifestation of cancer-induced immune suppression driving tumour progression.     

Prognostic significance of K-ras,p53, bcl-2, PCNA,CD34 in radically resected non-small cell lung cancers

The aim of this study was to investigate the prognostic significance of a panel of biological parameters in patients with radically resected non-small cell lung cancers (NSCLC). 269 cases with pathological stage I-IIIA NSCLC were retrospectively analysed. Immunohistochemistry was performed to detect protein expression of p53, bcl-2, proliferating cell nuclear antigen (PCNA) and CD34. Polymerase chain reaction (PCR)/direct nucleotide sequencing method was used to detect mutations in K-ras (codons 12, 13, 61, exons 1-2). The Kaplan-Meier estimates of survival were calculated for clinical and biological variables using the Cox model for multivariate analysis. Histological subtype and the pathologic tumour extension (pT) were the most powerful clinical-pathological prognostic factors for survival (P=0.030 and P=0.031, respectively), whereas among the biological parameters, p53 overexpression (P=0.032) and K-ras mutation (P=0.078) had a negative prognostic role, as demonstrated by multivariate analysis. Conversely, bcl-2, PCNA and CD34 expression were not correlated with survival. Statistically significant associations between p53 expression and the squamous cell carcinoma (SCC) subtype, bcl-2 expression and SCC subtype, K-ras mutation and p53 negative expression, p53 and bcl-2, bcl-2 and PCNA overexpression were observed. In conclusion, some biological characteristics such as the K-ras and p53 status may provide useful prognostic information in resected NSCLC patients, in addition to the classical clinico-pathological parameters. However, further studies are needed to clarify the value of adopting biological prognostic factor into clinical practice.     

Prognostic value of p27Kip1 and p21WAF/Cip protein expression in muscle invasive bladder cancer

Two genes, namely p27Kip1 and p21WAF/Cip1 that reveal distinct structural homology, have been identified as inductors of cell cycle arrest at the G1-checkpoint to prevent entry of somatic cells into the S phase of the cell cycle when substantial DNA damage has occurred. It was demonstrated that the p21WAF/Cip1 gene is induced by pathways dependent and independent from a functionally intact p53 tumour suppressor protein. It has been suggested that decreased expression both of the p21WAF/Cip1 and p27Kip1 protein may contribute to the development of human malignancies due to loss of critical antiproliferative mechanisms. So far, the role of altered p21WAF/Cip1 and mainly of a decreased p27Kip1 protein expression in patients with muscle invasive bladder cancer has not been investigated. In the present study, 50 tumour specimens from 50 patients undergoing radical cystectomy (T2-T4) were investigated for different biological and clinical characteristics as possible prognostic factors: age, depth of tumour infiltration (T-stage), histological grading (G), lymph node status as well as immunohistochemical staining for the p21WAF/Cip1 and p27Kip1 proteins. The median recurrence-free survival for patients with and without retained p21WAF/Cip1 protein expression was 54 months (3-86 months) and 13 months (1-40 months), respectively (p=0.07). During univariate analysis, loss of p21WAF/Cip1 protein expression (p=0.02), T-stage (p=0.02) and histological grading (p=0.03) were significant prognostic factors for survival, among which a negative reaction for the p21WAF/Cip1 protein (p=0.02) as well as T-stage (p=0.005) remained independent significant predictors during multivariate analysis. Loss of p27Kip1 protein expression was not correlated with the recurrence-free or the overall survival of the patients. Prospective studies are needed to confirm the independent prognostic potential of cell-cycle associated proteins such as p21WAF/Cip1 in patients with muscle invasive bladder cancer. The availability of more refined prognostic factors should assist decision making regarding the value of more aggressive treatment options, such as adjuvant or neoadjuvant chemotherapy, for defined subgroups of patients.     

The prognostic value of c‐Kit,K‐ras codon 12, and p53 codon 72 mutations in Egyptian patients with stage II colorectal cancer

BACKGROUND:

The prognosis for patients with colorectal cancer (CRC) depends mainly on standard clinicopathologic factors. However, patients with similar disease characteristics exhibit various outcomes, especially in stage II. Therefore, the identification of molecular prognostic markers is needed to predict patient outcomes.

METHODS:

The authors assessed the prognostic value of c‐Kit (also called cluster of differentiation 117 [CD117] or KIT), cyclooxygenase‐2 (COX‐2), tumor protein 53 (p53), and Kirsten rat sarcoma viral oncogene homolog (K‐ras) aberrations in 90 patients with stage II CRC using immunohistochemistry and molecular techniques. The results were correlated with standard clinicopathologic prognostic factors, overall survival (OS), and disease‐free survival (DFS).

RESULTS:

COX2 and c‐Kit overexpression was detected in 54.6% and 59.3% of patients, respectively. Overexpression of p53 was detected in 47 patients, including 29 who had mutations, and a unique mutation pattern was detected with 3 hotspots at codons 72, 245, and 273. Overexpression of ras was detected in 44 patients, including 37 who had mutations. On multivariate analysis, c‐Kit overexpression, p53 codon 72 mutations, perforation, and performance status were independent prognostic factors for DFS (P = .054, P = .015, P < .0001, and P = .043, respectively); whereas codon 12 K‐ras mutation, performance status, and perforation were independent prognostic factors for OS (P = .033, P = .006, and P < .0001, respectively).

CONCLUSIONS:

The current results provide evidence for the prognostic value of c‐Kit overexpression in patients with stage II CRC. The high p53 mutation rate and the unique hotspot in codon 72 have not been reported previously in CRC. This may be related to environmental or racial features that are unique to the studied population. Cancer 2010. © 2010 American Cancer Society.     

Down-regulation of tumor suppressor gene PTEN, overexpression of p53, plus high proliferating cell nuclear antigen index predict poor patient outcome of hepatocellular carcinoma after resection

We aimed to evaluate the interaction of two tumor suppressor genes PTEN and p53 and their relationship with cell cycle protein proliferating cell nuclear antigen (PCNA) in hepatocellular carcinoma (HCC). A total of 124 resected HCC paraffin specimens were collected from 1987 to 1999 for immunohistochemistry. Expression of PTEN, p53 and PCNA in HCC were analyzed for clinicopathologic correlation. The study revealed decreased or absent PTEN immunostaining (PTEN down-regulation) in 42.7% and positive p53 (p53+) immunostaining in 41.9% of HCC. There was a positive correlation between PTEN down-regulation and p53 (+) (P=0.001). PTEN down-regulation or p53 (+) correlated with increased HCC dedifferentiation, advanced pathologic stages and high PCNA labeling index (LI) of tumors (P<0.05). Patients with either PTEN down-regulation, p53 (+), or high PCNA LI had shorter survival and higher recurrence rates than patients with intact PTEN expression, p53 (-), or low PCNA LI respectively (P<0.05). By combining the three genes, patients with all PTEN down-regulation (+)/p53 (+)/high PCNA LI had the shortest overall survival (P<0.001) and the highest recurrence rates (P<0.001), followed by patients with two, one and none of three events accordingly. Combination of PTEN/p53/PCNA represented an independent prognostic factor for tumor recurrence and disease-specific survival (P<0.05). In conclusion, the down-regulated PTEN expression and p53 over-expression are involved in the pathogenesis of HCC. They correlate with high PCNA expression, HCC de-differentiation and advanced HCC stages. A combination of the three genes predicts patient outcome more powerfully than any of the individual genes.     

Correlation and expression of p53, HER-2, vascular endothelial growth factor (VEGF), and e-cadherin in a high-risk breast-cancer population

Background Many genetic traits common to aggressive breast carcinoma have been identified; yet little is known about the interrelationships of such traits during tumor development, especially in women prone to aggressive cancer. This study examined the expression of four biological markers associated with poor prognosis at each stage of breast cancer progression in primary tumors from women of lower economic status and assessed the relationship between these markers.Methods Archived primary breast tumors from 77 patients were assessed by immunohistochemical analysis for expression of human epidermal growth receptor 2 (HER-2), p53, vascular endothelial growth factor (VEGF), and e-cadherin, and the relationships between the expressions of these molecules were studied.Results Twenty-two (29%) patients had advanced (stage III or IV) disease. HER-2, VEGF, e-cadherin, and p53 signal were positive for 31 (40%), 58 (75%), 63 (82%), and 37 (48%) of patients, respectively. Among the markers tested, only p53 exhibited a significant association between expression and stage of the disease (P = 0.012). Expression of e-cadherin was positively associated with HER-2 overexpression (P = 0.004), and high levels of HER-2 occurred with strongly positive e-cadherin tumors. Marginally significant positive associations were observed between HER-2 and p53 signal (P = 0.06), and between disease stage and e-cadherin expression (P = 0.08).Conclusion The significant tendency toward expression of e-cadherin in conjunction with HER-2 overexpression in breast cancer is a novel finding. The association of p53 with more advanced stages of cancer emphasizes it as a key participant in metastatic processes in breast cancer.     

Integrated p53 histopathologic/genetic analysis of premalignant lesions of the esophagus

Esophageal carcinoma frequently occurs in patients with long-standing achalasia. AIM: To examine the role of p53 alterations and PCNA in patients with megaesophagus. METHODS: Sections of four tumors, and corresponding adjacent areas, from patients with achalasia due to Chagas' disease were examined by immunohistochemistry for p53 and PCNA proteins. Furthermore, 128 biopsies from 16 advanced achalasic patients were prospectively collected and evaluated for grades of inflammation, hyperplasia, dysplasia and also for p53 and PCNA proteins. All specimens showing p53 immunoreactivity were topographically genotyped using microdissection, PCR amplification and direct sequencing of p53 exons 5-8. RESULTS: Diffuse strong immunoreactivity of p53 was observed in 2/4 tumors. In one patient, the adjacent mucosa also showed strong p53. In the adjacent mucosa, the same areas showing p53 overexpression also had PCNA positive cells. In the prospective group, 7/16 (43.7%) patients or 53/128 (41.4%) biopsies expressed p53. The grade of inflammation was significantly correlated with the presence of positive p53, in patients, p = 0.004 and in biopsies, p < 0.00001. PCNA expression was found in the basal layer of the mucosa, and increased PCNA was associated with p53 overexpression, p = 0.00018. Genotyping detected mutation in exon 6, codon 213 RG, in one patient (1/16, 6.2%). CONCLUSIONS: (1.) p53 alterations, overexpression and mutational change, are an early event in patients with achalasia; (2.) The inflammation frequently seen in these patients appears to be associated with alterations of the p53 protein; (3.) Expression of the tumor suppressor gene is increased in areas showing proliferation.     

The prognostic value of MDM2 overexpression in superficial urothelial bladder carcinoma

The prognostic significance of p53 accumulation and MDM2 overexpression in superficial and papillary urothelial cell carcinoma (UCC) of the bladder has not yet been established. Therefore, MDM2 and p53 protein were investigated focusing on tumour grade and muscularis mucosae invasion in order to identify their prognostic significance. Thirty-five archival UCC were studied by immunohistochemistry using monoclonal antibodies DO7 against p53, and IB10 against MDM2. MDM2 overexpression was observed in 51.4% of the cases. The recurrence rate for patients with MDM2 overexpression was 7.09 fold higher than for those without MDM2 overexpression. p53 accumulation was not related to prognosis. There was no significant difference between low and high grade tumours in regard to MDM2 overexpression and p53 accumulation. pTa cases presented the higher frequency of MDM2 positive cases (66.7%) and a lower frequency of p53 accumulation (33.3%). MDM2 overexpression does not seem to be related to muscular mucosae invasion. Thus, MDM2 status may well be a prognostic marker in superficial urothelial bladder carcinomas.     

p53 expression predicts dismal outcome for medulloblastoma patients with metastatic disease

Medulloblastoma (MB) is the most common malignant primary brain tumour in childhood. Metastatic disease (M+) at diagnosis is the most important negative prognostic clinical marker and, despite craniospinal irradiation and intensive chemotherapy, it remains one of the leading causes of treatment failure. To date, few clinical and biological data have been evaluated to obtain an additional prognostic profile for these high-risk patients. In this study, 169 patients with metastatic MB registered in the multicentre HIT2000 trial of the German Society of Pediatric Oncology and Haematology (GPOH) have been investigated to determine the importance of p53 protein expression in predicting survival. At a median follow-up of 4.1 years, 159 patients with p53-negative tumours had significantly better four-year event-free survival (EFS) and progression-free survival (PFS) (56 ± 11, 59 ± 4%) than 10 patients with p53-positive tumours (40 ± 16, 40 ± 16%; P = 0.018 for EFS, P = 0.007 for PFS, respectively). Furthermore, four-year overall survival (OS) of children with p53-negative tumours was higher than for children with p53-positive tumours (72 ± 4 vs. 35 ± 18%, P = 0.05). Three of the p53-positive MBs harbored a point mutation in the TP53 gene. p53 protein assessment by immunohistochemistry may be a useful tool for sub-stratification of metastatic high-risk MB patients.     

P53 nuclear overexpression and disease progression in ta-bladder carcinoma

This study investigates the prevalence and clinical relevance of nuclear overexpression of the p53 protein, as detected by antibody PAb1801, in 54 patients with pathologically confirmed Ta bladder carcinomas obtained by transurethral resection at Memorial Sloan-Kettering Cancer Center between 1972 and 1980. No patient received any prior adjuvant therapy. Clinical and histopathological prognostic variables such as age, sex and tumor grade were also analyzed. The median follow-up was 110 months. Patients were stratified into two groups according to the percent of tumor cells with nuclear overexpression of the p53 protein. Group A included carcinomas with less than 20% (n=42) and Group B with 20% or more (n=12) of tumor cells with positive nuclear immunoreactivity. Five patients of Group A (12%) developed tumor progression as did 7 (58%) patients of Group B (p=0.002). Three (7%) patients of Group A and 3 (25%) patients of Group B died of bladder cancer (p=0.12). This study demonstrates a relatively low prevalence of altered p53 expression in this early stage of bladder cancer (22%). Nuclear overexpression of the p53 protein in greater-than-or-equal-to 20% tumor cells,was highly associated with tumor grade (p=0.01), but the former was the only independent marker of tumor progression (p=0.0008) on the basis of the multivariate analysis performed. It was also the only independent variable . associated with death due to bladder cancer (p=0.04). We conclude that p53 nuclear overexpression is a prognostic indicator in this disease, and may be useful for selection of therapy for patients with non-invasive papillary superficial bladder carcinoma.     

肺癌p53蛋白和增殖细胞核抗原的检测及其意义

目的：探讨ｐ５３蛋白、ＰＣＮＡ联合表达在临床中的意义。方法应用免疫组织化学Ｓ－Ｐ法检测１０１例肺癌中ｐ５３蛋白、ＰＣＮＡ的表达。结果ｐ５３蛋白在肺癌组织中有较高的表达。总阳性率为５９．４％（６０／１０１）。Ｐ５３蛋白表达与肿瘤分化程度呈正相关（Ｐ〈０．０５）。而与肺癌患者的生存期呈负相关（Ｐ〈０．０１）。Ｐ５３蛋白表达与ＰＣＮＡ染色密切相关（Ｐ〈０．０５）。结论Ｐ５３蛋白表达可作为判断肺癌患者预后     

Combination of molecular alterations and smoking intensity predicts bladder cancer outcome

BACKGROUND:

Traditional single‐marker and multimarker molecular profiling approaches in bladder cancer do not account for major risk factors and their influence on clinical outcome. This study examined the prognostic value of molecular alterations across all disease stages after accounting for clinicopathological factors and smoking, the most common risk factor for bladder cancer in the developed world, in a population‐based cohort.

METHODS:

Primary bladder tumors from 212 cancer registry patients (median follow‐up, 13.2 years) were immunohistochemically profiled for Bax, caspase‐3, apoptotic protease‐activating factor 1 (Apaf‐1), Bcl‐2, p53, p21, cyclooxygenase‐2, vascular endothelial growth factor, and E‐cadherin alterations. “Smoking intensity” quantified the impact of duration and daily frequency of smoking.

RESULTS:

Age, pathological stage, surgical modality, and adjuvant therapy administration were significantly associated with survival. Increasing smoking intensity was independently associated with worse outcome (P < .001). Apaf‐1, E‐cadherin, and p53 were prognostic for outcome (P = .005, .014, and .032, respectively); E‐cadherin remained prognostic following multivariable analysis (P = .040). Combined alterations in all 9 biomarkers were prognostic by univariable (P < .001) and multivariable (P = .006) analysis. A multivariable model that included all 9 biomarkers and smoking intensity had greater accuracy in predicting prognosis than models composed of standard clinicopathological covariates without or with smoking intensity (P < .001 and P = .018, respectively).

CONCLUSIONS:

Apaf‐1, E‐cadherin, and p53 alterations individually predicted survival in bladder cancer patients. Increasing number of biomarker alterations was significantly associated with worsening survival, although markers comprising the panel were not necessarily prognostic individually. Predictive value of the 9‐biomarker panel with smoking intensity was significantly higher than that of routine clinicopathological parameters alone. Cancer 2013. © 2013 American Cancer Society.     

膀胱移行细胞癌中p53突变、bcl-2和PCNA表达的临床意义

To correlate the frequency of p53 mutations, bcl-2 expression and the proliferation status (proliferating cell nuclear antigen, PCNA) in patients with bladder cancer with cell proliferation, apoptosis and their clinico-pathologic findings. Paraffin-embedded sections from 39 superficial (T1G1-G3) and 23 invasive (T2-T4a G3 N0M0) primary transitional cell carcinomas (TCC) in the bladder were investigated immunohistochemically for p53, bcl-2 and PCNA. The median follow-up was 37 months; 24 had recurrences. The proliferation index (PI) was expressed as a percentage of the PCNA-positive cells in the tumor cells. Apoptosis was detected by terminal deoxy-nucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and the apoptotic index (AI) was expressed as a percentage of the TUNEL-positive tumor cells. p53 mutation was identified in 50 patients (80.6%). The mutation was most common in tumors grade 3 (91.3%) as compared to grade 2 (78.5%) and grade 1 (72.7%, P<0.05). Stage pT2 tumors had a higher frequency of p53 mutation (95.7%) as compared to pTa-1 tumors (74.3%, P<0.01). Only 14 tumors (22.5%) expressed bcl-2; grade 3 tumors expressed bcl-2 significantly more frequently (P<0.05); there was no correlation between bcl-2 and tumor stage. There was no interrelation between p53 mutation and bcl-2 expression (P>0.05). The PI ranged from 17.2% to 41.8% (median 22.4%) and the AI from 1.9% to 3.5% (median 2.9%) in bladder cancer. Statistical analyses revealed a close associations between PI, AI and tumor grade and stage of bladder cancer. p53 mutation correlates with invasion. p53 and PCNA overexpression may offer valuable additional prognostic information in bladder tumors. With the progression of the tumor grade, cell proliferation may be accompanied by frequent apoptosis in bladder cancer, but the PI increased much more than the AI.     

Prognostic value of the apoptotic index analysed jointly with selected cell cycle regulators and proliferation markers in non-small cell lung cancer

In a previous small series of surgically treated non-small cell lung cancer patients (NSCLC), we found that higher apoptotic index (AI) negatively influenced survival (Dworakowska D, Jassem E, Jassem J, Karmolinski A, Dworakowski R, Wirth T, et al. Clinical significance of apoptotic index in non-small cell lung cancer: correlation with p53, mdm2, pRb and p21WAF1/CIP1 protein expression. J Cancer Res Clin Oncol 2005; 131:617–623.). In this study we attempted to verify our previous finding in larger group of 170 NSCLC cases, additionally correlating AI to selected cell cycle regulators as well as a proliferation marker. Apoptosis was assessed with the use of the TUNEL technique, whereas the expression of p53, pRb, mdm2, p21^WAF1/CIP1, cyclin D1 and PCNA were assessed immunohistochemically. The mean and the median AI was 12 and 8, respectively. The expression of p53, pRb, mdm2, p21^WAF1/CIP1 proteins and cyclin D1 was found in 47%, 71%, 37%, 65% and 40% of cases, respectively. The mean and the median PCNA labeling index (PCNA LI) was 34 and 35, respectively. AI was not correlated with any patient characteristic or other tumor markers. In uni- and multivariate analysis AI, analysed separately or jointly with cell cycle regulators and PCNA LI, did not influence disease-free or over-all survival. However, patients with “very high AI/very high PCNA LI” had a particularly poor prognosis (P = 0.001). Patients with “very low AI/negative pRb” phenotype survived for a shorter time in comparison to others (P = 0.04). In addition, patients with the highest PCNA LI had a worse outcome in comparison to patients with the lowest PCNA LI (P = 0.04), especially those with concomitant p53 protein expression (P = 0.026) or lacking pRb protein expression (P = 0.04). This study demonstrates that joint analysis of several factors involved in apoptosis, proliferation and cell cycle regulation, but not AI alone, might provide additional prognostic information in NSCLC patients.     

Relationship between p53 gene mutation and protein expression: clinical significance in transitional cell carcinoma of the bladder

Although the mutated p53 gene has been postulated to induce immunohistochemically-detectable p53 protein, reports regarding the relationship between p53 mutation and p53 protein expression have been contradictory. This study investigated the relationship between p53 mutations and p53 expression and their clinical significance for patients with transitional cell carcinoma of the bladder. Eighty-seven transitional cell carcinoma of the bladder were analyzed by immunohistochemistry (IHC) for p53 nuclear accumulation, and the results compared to mutations detected in the p53 gene evaluated by polymerase chain reaction single-strand conformation polymorphism (SSCP) and DNA sequence analysis. By p53 IHC analysis, positive p53 staining was observed in 50 (57.5%) of the 87 tumors. The specificity of IHC, defined as a percentage of IHC negative (<20%) tumors among tumors without mutation, was 94.6%. Despite the good concordance between p53 mutation and p53 protein expression (p<0.0001), 48.0% (24/50) of the tumors showed p53 overexpression without mutation, and 2 (5.4%) tumors with mutation showed no p53 immunoreactivity. Patients with higher grade (grade 3), stage (stages pT2-4), and p53 mutations had a poorer prognosis by Kaplan-Meier survival analysis. A Cox univariate analysis found that grading (hazard ratio 3.139; p=0.002), staging (hazard ratio 3.832; p=0.0005) and p53 mutation (hazard ratio 2.498; p=0.013) were significant variables in these patients, but no variable was independently associated with an increased survival of bladder carcinoma by multivariate analysis. We found that a 20% cut-off level of p53 overexpression showed the highest correlation with prognosis and p53 mutation, however, p53 overexpression and mutation were not superior to staging as prognostic markers. These data suggest that careful assessment of the TNM staging system remains the most reliable predictive indicator of survival for patients with transitional cell carcinoma of the bladder.