Over-expression of S100B protein as a serum marker of brain metastasis in non-small cell lung cancer and its prognostic value

Validated serum biomarkers for patients suffering from non-small cell lung cancer (NSCLC) brain metastasis are urgently needed for early diagnosis, treatment monitoring, and prognostic classification in daily clinical practice. Serum S100B was reported to be a marker of leaky blood-brain barrier (BBB), which was often caused by brain tumors. This study aimed to investigate the role of S100B in NSCLC brain metastasis. The results showed that serum S100B correlated significantly with NSCLC brain metastasis (P?<?0.001). When evaluated by the ROC curve, at the cutoff point 13.83 pg/ml, the sensitivity and specificity were 94% and 93%, respectively (AUC= 0.938, P < 0.001). High level of serum S100B was significantly correlated with a higher number of brain metastases and significantly worse prognosis (P?<? 0.05). In addition, S100B was an independent prognostic factor (P?<? 0.001). In conclusion, serum S100B was a sensitive and specific marker for early detection of brain metastasis in NSCLC and could be used as a surveillance tool for prognosis evaluation.     

Immunohistochemical analysis of a muscle ankyrin-repeat protein, Arpp, in paraffin-embedded tumors: evaluation of Arpp as a tumor marker for rhabdomyosarcoma

Arpp, a protein including an ankyrin-repeat, P EST motif, and p roline-rich region, is a recently identified protein that is exclusively expressed in striated muscles. This study comprehensively analyzed its expression among soft tissue sarcomas of various histological types and evaluated its potential use for the differential diagnosis of rhabdomyosarcoma (RMS). Formalin-fixed, paraffin-embedded tissues, including 37 RMS cases, 88 non-RMS sarcomas, and 38 carcinomas, were analyzed for Arpp expression. Arpp was detected in 33 (89.2%) of 37 RMS cases by immunohistochemistry. Western blot analysis revealed expression of Arpp in all RMS cases tested. High expression of Arpp was generally associated with morphological evidence of skeletal muscle differentiation of tumor cells. In contrast, Arpp displayed 6.3% (8/126) positivity among the non-RMS tumors. Focal or weak expression was seen in malignant fibrous histiocytoma (2/27), synovial sarcoma (1/11), Ewing sarcoma (1/5), and epithelioid sarcoma (3/5), whereas one epithelioid sarcoma displayed strong expression for Arpp. A comparative analysis of the Arpp profile with that of myogenic markers in RMS revealed that the sensitivity of Arpp (89.2%) was higher than that of myoglobin (59.6%) and comparable with that of myogenin (88.2%), MyoD (80.6%), muscle-specific actin (83.8%), and desmin (89.2%). These results suggested that Arpp is sensitive to and specific for RMS. Thus, we proposed that Arpp is a novel skeletal muscle-specific marker, which is useful for differential diagnosis of RMS.     

Limited value of KAI1/CD82 protein expression as a prognostic marker in human gastric cancer

The cell surface glycoprotein KAI1/CD82 suppresses tumor growth and metastasis in animal models. This study aimed to evaluate the prognostic relevance of KAI1/CD82 protein expression in human gastric cancer. Primary gastric carcinomas (n=271) with a mean clinical follow-up time of 48 months were immunostained using the monoclonal anti-KAI1/CD82 antibody G2. Staining was evaluated as negative versus positive for statistical analysis. KAI1/CD82 immunoreactivity was absent in 103/271 (38%) cases. There was a trend towards KAI1/CD82 negativity in poorly differentiated cases (p=0.0679). Moreover, KAI1/CD82-negative carcinomas were associated with a higher pT status (p=0.0222), metastatic lymph node involvement (p=0.0018) and a higher clinical tumor stage (p=0.0050). The median overall survival times of KAI1/CD82-negative and KAI1/CD82-positive gastric carcinomas were 20 and 37 months, respectively (p=0.2305). These results are in line with the proposed function of KAI1/CD82 as a suppressor of tumor growth and metastasis. However, these data suggest that KAI1/CD82, as detected by immunohistochemistry, is of limited value as a prognostic marker for gastric cancer in routine histological workup.     

Skp2与肿瘤的生物学性状、预后及治疗

S期激酶相关蛋白2(S-phase k inase assoc iated prote in 2,Skp2)是泛素连接酶复合物的底物识别序列,对正常细胞周期调控起重要作用。在多数恶性肿瘤组织中Skp2呈过表达,并通过泛素化降解细胞周期蛋白依赖性激酶抑制剂p27k ip1,使细胞G1-S期进程加速。Skp2具有致癌潜能,与肿瘤的增殖,分化,抵抗凋亡和耐药性等生物学性状关系密切。Skp2不仅可作为判断肿瘤患者预后独立的指标,还可能成为肿瘤的药物和基因治疗的新靶点。     

细胞周期蛋白D1、Ki-67和bcl-2的表达与CD117阳性的胃肠道间质瘤生物学行为的关系

目的 检测细胞周期蛋白D1（cyclin D1）、Ki-67和bcl-2在胃肠道间质瘤（GIST）中的表达,探讨它们在GIST发生、发展中的作用及临床病理意义。方法 59例手术切除GIST标本进行CD117、CD34、平滑肌肌动蛋白（SMA）、结蛋白、S-100、cyclin D1、bcl-2和Ki-67免疫组织化学染色,同时进行病理形态学观察包括形态学类型、肿瘤大小、坏死和核分裂象。所有病例随访2～9年。所有数据进行单因素、多因素和相关分析。结果 随访40例患者一直健在,15例患者死于GIST,4例患者死于其他原因。统计学分析显示肿瘤直径〉5cm、有坏死、核分裂象在每50个高倍视野〉5个、Ki-67增殖指数（LI）〉5％、cyclin D1和bcl-2免疫组织化学染色强阳性都可以作GIST患者手术后的预测指标,且具有统计学意义;核分裂象和Ki-67增殖指数是独立的预测指标;Ki-67 LI≥5％和核分裂象≥5／50 HPF呈正相关（r=0．532,P〈0．01）;cyclin D1与bcl-2强阳性表达呈正相关（r=0.273,P〈0．05）。结论 肿瘤大小、坏死、核分裂象、cyclin D1、Ki-67增殖指数和bcl-2可作为GIST患者临床预测指标;核分裂象和Ki-67增殖指数可作为独立的预测指标;cyclinD1与bcl-2呈明显相关性,Ki-67免疫组织化学染色可以代替核分裂象作为一项很有用的预测指标。     

Clinical significance and prognostic value of chromatin assembly factor-1 overexpression in human solid tumours

Polo S E, Theocharis S E, Grandin L, Gambotti L, Antoni G, Savignoni A, Asselain B, Patsouris E & Almouzni G
(2010) Histopathology 57 , 716–724
Clinical significance and prognostic value of chromatin assembly factor‐1 overexpression in human solid tumours Aims: Chromatin assembly factor‐1 (CAF‐1), whose function is critical for maintaining chromatin stability during DNA replication and repair, has been identified as a proliferation marker in breast cancer. The aim was to investigate CAF‐1 as a proliferation marker in a wide variety of solid tumours, and to assess its potential value in predicting clinical outcome. Methods and results: Using immunocytochemistry on paraffin‐embedded tissue sections, the CAF‐1 labelling index was compared with known proliferation markers Ki‐67 and minichromosome maintenance (MCM), and its association with clinicopathological data and patients’ outcome analysed. CAF‐1 expression showed a strong positive correlation with Ki‐67, used routinely to detect proliferating cells, while it generally displayed weaker correlations with MCM markers, known to label cells with replicative potential. CAF‐1 expression was associated significantly with histological grade in breast, cervical, endometrial and renal cell carcinomas, and with disease stage in endometrial and renal carcinomas. Furthermore, high expression of CAF‐1 was an independent predictor of adverse clinical outcome in renal, endometrial and cervical carcinomas. Conclusions: CAF‐1 is a proliferation marker in various malignant tumours with prognostic value in renal, endometrial and cervical carcinomas, which supports the value of CAF‐1 as a clinical marker of cancer progression.     

人类额外小标记染色体的鉴定及其研究价值的探讨

目的 应用荧光原位杂交(fluorescent in situ hybridization,FISH)结合染色体核型分析了解人类额外小标记染色体(small supernumerary marker chromosomes,sSMC)的来源,并探讨其发生机理及应用价值.方法 对3例羊水染色体核型分析结果显示是47,XN,+mar的胎儿羊水细胞用两种探针cepFISH(针对着丝粒)和SubcenM-FISH(针对近着丝粒区域)进行分析.结果 病例1的FISH结果为47,XY,+mar.ish inv dup(22) (q11.1)(D22Z4++,D14/22Z1+,RP11-172D7-),标记染色体完全由异染色质组成,胎儿活产未见异常临床表型.病例2的FISH结果为47,XX,+mar.ish r(10) (p11.2q11.2)(cep10+,RP11 232C13+,RP11-178A10+)[25]/46,XX[10],标记染色体由着丝粒附近的常染色质和异染色质组成,胎儿活产无异常临床表型.病例3的FISH结果为47,XY,+mar.ish inv dup(22) (q11.1)(D22Z4+,D14/22Z1+),标记染色体由异染色质组成,胎儿B超有异常但与此标记染色体关系不明.结论 由于sSMC来源的多样性,给产前诊断带来了巨大的困难.其鉴定需要在传统的显带核型分析基础上结合FISH或者其他分子技术.其特殊的结构也为基因定位、异染色质研究以及基因治疗等提供了非常有价值的研究载体.     

套细胞淋巴瘤的临床病理特征及预后因素分析

目的探讨套细胞淋巴瘤（MCL）的临床病理特征及预后因素。方法对102例经形态学及免疫表型检测确定的MCL进行分析,组织病理制片和链霉素抗生物素蛋白过氧化物酶法或EnVision法染色,并进行了随访。结果102例患者中位年龄59岁（30～79岁）,男女之比约2．92：1。淋巴结是最常受累的部位（98／98,100％）,结外常受累的部位：骨髓（29／45,64．4％）、脾脏（36／57,63．2％）、咽淋巴环（15／48,31．3％）、外周血（15／51,29．4％）、肝脏（12／53,22．6％）及胃肠道（15／102,14．7％）;87．7％（71／81）初次就诊时处于临床Ⅲ～Ⅳ期,45．5％（25／55）患者有B症状;48．7％（19／39）患者血清乳酸脱氢酶升高。除7例（6．9％）因组织取材小无法区分病变模式外,余95例中12例（11．76％）为套区增生型,41例（40．2％）结节型,42例（41．2％）弥漫型。75．5％（77）经典型,24．5％（25例）瘤细胞呈母细胞样变型。102例均表达B细胞标记而不表达T细胞标记,96例（94．1％）肿瘤细胞表达细胞周期蛋白D1,70例（71．4％）CD5弱阳性。68例获得随访,中位生存时间10个月（0～89个月）。套区增生型＋结节型、经典型的核分裂象≤15／10HPF,增殖指数≤15％;骨髓无受累,提示患者预后好,而其他临床病理因素对患者生存未见影响。结论国内MCL患者病征与国外患者基本一致,其病变模式、细胞变型、核分裂象、增殖指数、骨髓是否受累及受累程度与预后有关。     

Diagnostic and prognostic value of ARID1A in endometrial hyperplasia: a novel marker of occult cancer

AT‐rich interaction domain 1A (ARID1A) is a tumor suppressor protein involved in endometrioid carcinogenesis. The expression of ARID1A may be lost in the premalignant phase. Our aim was to assess ARID1A as: (i) diagnostic marker to differentiate premalignant endometrial hyperplasia (EH) form benign EH; (ii) prognostic marker for the risk of occult cancer in premalignant EH. A systematic review and meta‐analysis were performed by searching electronic databases from their inception to October 2018 for all studies assessing ARID1A in EH by immunohistochemistry. Sensitivity, specificity, positive and negative likelihood ratios (LR+ and LR?), and diagnostic odds ratio (DOR) were calculated for both diagnostic and prognostic accuracy. LR+ > 5, LR? < 0.2, DOR > 25 defined good accuracy; LR+ > 10, LR? < 0.1, DOR > 100 defined excellent accuracy. Seven studies with 467 EH were included. As diagnostic marker, ARID1A showed sensitivity = 0.12, specificity = 0.99, LR+ = 4.34, LR? = 0.85, DOR = 5.12. As prognostic marker for occult cancer, ARID1A showed sensitivity = 0.33, specificity = 0.99, LR+ = 20.70, LR? = 0.49, DOR = 49.59. In conclusion, ARID1A loss is highly specific, but little accurate as diagnostic marker of premalignant EH. Instead, ARID1A loss in premalignant EH is an accurate and almost perfectly specific prognostic marker for coexistent cancer.     

Elevated content of p53 protein in the absence of p53 gene mutations as a possible prognostic marker for human renal cell tumors

P53 tumour suppressor gene expression was estimated immunohistochemically using DO-1 monoclonal antibody (recognising both wild-type and mutant p53) in 88 human renal tumours. Single strand conformation polymorphism (SSCP) analysis of possible mutations within exons 4–8 of the p53 gene was performed in 29 of the tumours (mostly immunostaining-positive cases). Obviously elevated p53 content was detected with DO-1 antibody in chromophobic cell carcinomas and most clear/chromophilic cell tumours (in chromophilic cell populations). In contrast, clear cell carcinomas demonstrated either complete absence of p53 expression or the presence of single immunopositive nuclei. Oncocytomas were completely negative. Additional immunostaining of the positive samples with mutant p53-specific Pab240 monoclonal antibody failed to detect immunopositive material. No p53 mutation was found in any of the samples analysed by SSCP. Our results suggest that the elevated p53 content in human renal cell carcinomas does not result from gene mutation and that p53 gene alterations are probably not an important mechanism in the development of human renal cell carcinomas. Accumulation of the wild-type p53 protein may be a useful prognostic marker indicating neoplastic progression and malignancy.     

Clinicopathologic and immunohistochemical correlation in sporadic pancreatic endocrine tumors: possible roles of utrophin and cyclin D1 in malignant progression

Pancreatic endocrine tumors (PETs), both functioning and nonfunctioning, are usually well differentiated and progress slowly. The 2004 World Health Organization (WHO) criteria classify PETs according to clinicopathologic features and Ki-67 proliferative index. A tumor associated with poorer prognostic features may be considered "uncertain" in behavior, but the malignant classifications are reserved for tumors showing clear signs of aggressive behavior. It remains difficult to predict malignant progression in any individual PET. The cytoskeletal protein utrophin is encoded on chromosome 6q, a region frequently lost in malignant PETs. Cyclin D1 is a highly regulated mediator of the cell cycle and is frequently overexpressed in sporadic PETs. Sporadic PETs resected or biopsied from 40 patients were identified and classified using WHO criteria (19 benign/uncertain, 21 malignant). Distinctive patterns of biologic activity in unequivocally malignant PETs were demonstrated by immunohistochemistry for utrophin and cyclin D1. Utrophin localized to cell membranes (76% in malignant versus 21% in benign/uncertain PETs, P < .0006) and cyclin D1 staining showed nuclear positivity (67% in malignant versus 17% in benign/uncertain PETs, P < .003). Membranous utrophin localization was associated with significantly reduced patient survival (P = .045). Both membranous utrophin and nuclear cyclin D1 staining were also associated with higher Ki-67 proliferative indices. In our series, neither utrophin nor cyclin D1 was predictive of malignant progression in uncertain (WHO 1.2) PETs. Further studies are warranted to elucidate the role of utrophin and cyclin D1 in the malignant progression of PETs.     

Phosphorylated 4E-binding protein 1 (p-4E-BP1): a novel prognostic marker in human astrocytomas

Korkolopoulou P, Levidou G, El‐Habr E A, Piperi C, Adamopoulos C, Samaras V, Boviatsis E, Thymara I, Trigka E‐A, Sakellariou S, Kavantzas N, Patsouris E & Saetta A A
(2012) Histopathology  61, 293–305 Phosphorylated 4E‐binding protein 1 (p‐4E‐BP1): a novel prognostic marker in human astrocytomas Aims: To investigate the significance of the mammalian target of rapamycin (mTOR) pathway in astrocytic tumours, published information in this context being limited, especially regarding phosphorylated 4E‐binding protein (p‐4E‐BP) 1. Methods and results: Paraffin‐embedded tissue from 111 patients with astroglial tumours (grades II–IV) was investigated for the association of phosphorylated mTOR (p‐mTOR) signalling components with phosphorylated extracellular signal‐related kinase 1/2 (p‐ERK1/2) and phosphorylated AKT (p‐AKT) expression, clinicopathological features, angiogenesis, isocitrate dehydrogenase 1 (IDH1)‐R132H, and survival. Expression was also quantified by western blot analysis in 12 cases and in three primary glioma cell cultures following rapamycin treatment. p‐mTOR expression correlated with p‐4E‐BP1 expression and marginally with p‐p70S6K expression. p‐4E‐BP1 expression increased with tumour grade. Rapamycin induced a decline in phosphorylation levels of all three proteins. Nuclear p‐AKT and cytoplasmic p‐ERK1/2 immunoexpression correlated with p‐4E‐BP1 expression, whereas cytoplasmic p‐AKT expression correlated with p‐p70S6K expression. All three proteins were associated with increased angiogenesis but not with IDH1‐R132H expression status. p‐mTOR adversely affected overall and disease‐free survival in univariate analysis. In multivariate survival analysis, the presence of p‐4E‐BP1 predicted shortened overall survival in the entire cohort and glioblastomas. Conclusions: mTOR signalling components are differentially involved in the acquisition of a more aggressive and angiogenic phenotype in astrocytic tumours. Moreover, p‐4E‐BP1 emerges as a novel prognostic marker, which might aid in the selection of patients who are more likely to benefit from therapy with mTOR inhibitors.     

The cohesin-interacting protein, precocious dissociation of sisters 5A/sister chromatid cohesion protein 112, is up-regulated in human astrocytic tumors

Glioblastoma multiforme (GBM) is the most prevalent, highly malignant, invasive and difficult-to-treat primary brain tumor in adults. At the genetic level, it is characterized by a high degree of chromosomal instability and aneuploidy. It has been shown that defects in the mitotic spindle checkpoint could lead to the development of aneuploidy as well as tumorigenesis. Additional proteins regulating sister chromatid cohesion could also be involved in maintaining the fidelity of chromosome segregation. One such protein is the precocious dissociation of sisters 5A (Pds5A), also known as sister chromatid cohesion protein 112. It is a nuclear protein, expressed from the S right through to the mitotic phase. It is highly conserved from yeast to man and plays a role in the establishment, maintenance and dissolution of sister chromatid cohesion. The mutation of Pds5A orthologs in lower organisms results in chromosome missegregation, aneuploidy and DNA repair defects. It is considered that such defects can cause either cell death or contribute to the development of cancer cells. Indeed, altered expression levels of Pds5A have been observed in tumors of the breast, kidney, oesophagus, stomach, liver and colon. Malignant gliomas, however, have not been analysed so far. Herein, we report on the cloning of Rattus norvegicus Pds5A and on the analysis of its expression pattern in rat tissue. We also show that Pds5A is significantly overexpressed at both the mRNA and protein level and that this overexpression correlates positively with the WHO grade of human gliomas. However, functional assays show that the siRNA-mediated knockdown of Pds5A affects sister chromatid cohesion but does not influence mitotic checkpoint function or the proliferation and survival of GBM cells. Although the mechanism by which Pds5A functions in GBM cells remains unclear, its overexpression in high grade gliomas implies that it could play a pivotal role during the development and progression of astrocytic tumors.     

Chromosome DNA imbalances in human astrocytic tumors: A comparative genomic hybridization study of 63 Chinese patients

Astrocytic tumors are the most frequent primary brain neoplasms. They are clinically characterized by wide variations in histology. Analysis of chromosome DNA imbalance may help to advance diagnosis, grading, and classification, and to determine appropriate therapeutic approaches for tumors of astrocytic lineages. Comparative genomic hybridization (CGH) provides comprehensive information about chromosome DNA aberrations, and is an important technique for evaluating the differences at genomic levels among the same or different grade tumors. In this study, 63 astrocytic tumors of Chinese patients were screened by CGH, and the relationship between their chromosome DNA imbalances and the histopathological classification, grading, and clinical features was analyzed. Most tumors showed genomic copy aberrations detected by CGH. The most frequent abnormalities were regional gains in chromosome 1q and 7p; regional losses in chromosome 1p, 2q, 4q, 6p, 10q, 12q, 15q, 19q, and 22q were also frequently observed. The gain of 1q and the loss of 15q were relevant to the histological types and grades of WHO classification. The losses of 4q and 10q correlated with age in the group of anaplastic astrocytoma, which was unreported in the literature. This study confirmed that chromosomal aberrations, such as +1q, −4q, −10q, +7p, and −15q possibly contributed to the pathogenesis of these tumors. Our data was the first report on the chromosomal aberrations of astrocytic tumors of Chinese patients.     

Time to positivity in bloodstream infection is not a prognostic marker for mortality: analysis of a prospective multicentre randomized control trial

ObjectivesTime to positivity (TTP), calculated automatically in modern blood culture systems, is considered a proxy for microbial load and has been suggested as a potential prognostic marker in bloodstream infections. In this large, multicentre, prospectively collected cohort, our primary analysis aimed to quantify the relationship between the TTP of monomicrobial blood cultures and mortality.MethodsData from a multicentre randomized controlled trial (RAPIDO) in bloodstream infection were analysed. Bloodstream infections were classified into 13 groups/subgroups. The relationship between mortality and TTP was assessed by logistic regression, adjusted for site, organism, and clinical variables, and linear regression was applied to examine the association between clinical variables and TTP. Robustness was assessed by sensitivity analysis.ResultsIn total 4468 participants were included in the RAPIDO. After exclusions, 3462 were analysed, with the most common organisms being coagulase-negative staphylococci (1072 patients) and Escherichia coli (861 patients); 785 patients (22.7%) died within 28 days. We found no relationship between TTP and mortality for any groups except for streptococci (odds ratio (OR) with each hour 0.98, 95%CI 0.96–1.00) and Candida (OR 1.03, 95%CI 1.00–1.05). There was large variability between organisms and sites in TTP. Fever (geometric mean ratio (GMR) 0.95, 95%CI 0.92–0.99), age (GMR per 10 years 1.01, 95%CI 1.00–1.02), and neutrophilia were associated with TTP (GMR 1.03, 95%CI 1.02–1.04).ConclusionsTime to positivity is not associated with mortality, except in the case of Candida spp. (longer times associated with worse outcomes) and possibly streptococci (shorter times associated with worse outcomes). There was a large variation between median times across centres, limiting external validity.