基质金属蛋白酶和组织金属蛋白酶抑制剂在肾细胞癌组织中的表达

目的:基质金属蛋白酶及组织金属蛋白酶抑制剂在肾细胞癌转移中占有重要的作用,研究肾细胞癌组织中MMP-2、MMP-9、TIMP-1和TIMP-2的表达情况,为肾癌转移的治疗提供理论依据。方法:选取36例肾细胞癌肾组织标本,从相同的肾细胞癌组织及癌旁肾组织获得对照样本,均进行根治性肾切除手术切除。肿瘤分期按TNM分期标准。为了统计评估,肿瘤1期和2期为低级,3期以上为高级。RT-PCR检测肿瘤和正常组织中的MMP-2、MMP-9、TIMP-1和TIMP-2的表达。结果:不同样本MMPs和TIMPs表达水平各不相同。肾细胞癌组织中MMP-2、MMP-9、TIMP-1、TIMP-2在肾细胞癌中的表达明显高于正常肾组织(P0.05)。在肾细胞癌的肿瘤分期方面,MMP-2与MMP-9和肿瘤的分期显著相关,随着肿瘤分期的增加,MMP-2与MMP-9的表达明显升高(P0.05),而TIMP-1与TIMP-2与肿瘤的分期无关。结论:肾细胞癌组织中TIMP-2、MMP-2,MMP-9,TIMP-1的mRNA表达显著高于正常肾组织,抑制MMPS的表达将成为治疗肾细胞癌转移的新的方向。     

Proliferating cell nuclear antigen in normal urothelium and urothelial lesions of the urinary bladder: a quantitative assessment using a true color image analysis system

To evaluate proliferating cell nuclear antigen (PCNA) staining for assessing proliferative activity in routine pathology specimens of urinary bladder, the bladder carcinoma cell line J82 and a total of 122 specimens of normal bladder and urothelial lesions were stained with the antibody clone PC10 against proliferating cell nuclear antigen. In in vitro plateau cultures the proportion of PCNA-positive cells exceeded that of Ki-67-positive cells, and only very few cells were negative. In formalin-fixed tissues, the PCNA staining pattern, which should be confined to replicon units in the nucleus, was optimized by 1 h postfixation in an organic solvent (methacarn). Sections showed positive nuclear staining confined to basal and some suprabasal cells in normal urothelium and grade 1 dysplasias, but more generalized nuclear staining in all other neoplastic lesions. In addition, stromal cells adjacent to invasive tumors showed nuclear positivity in some instances. Using quantitative true color image analysis of sections counterstained with hemalum, the degree of brown staining of the PCNA reaction product is contrasted with the blue staining of the nuclear area. With this method low contrast specific staining not appreciated optically can be reliably detected. Image analysis data confirmed observations made on noncounterstained sections and showed significant differences between grade 1 and 2 dysplasias as well as between grade 1 dysplasia and all grades of papillary tumor. Furthermore, a significant difference in PCNA staining indices was found between grade 1 and 3 bladder carcinomas. The results indicate that PCNA staining using the PC10 antibody is not confined to the proliferative fraction of neoplastic urothelium. In contrast with data from normal tissue and malignant hematological neoplasms, the amount of PCNA is regulated differently in urothelial neoplasms, emphasizing the biological differences between the following two sets: mild dysplasia and moderate dysplasia; mild dysplasia and papillary carcinomas. The use of image analysis to standardize the detection process after controlled staining conditions is advisable in order to provide reliable data. Supported by the DFG project: Knuechel/Urothelcarcinom 263     

Postoperative bladder washing cytology after transurethral resection. Can it predict the recurrence of urothelial carcinoma?

OBJECTIVE: To assess the ability of postoperative bladder washing cytology, performed immediately after transurethral resection of mostly stage Ta or T1 papillary urothelial carcinoma, to predict early recurrence. STUDY DESIGN: In a 1-year period, preoperative and postoperative bladder washing cytology specimens were sampled from patients undergoing transurethral resections in which all visible tumor was removed. There were 38 resections in 32 patients. RESULTS: Postoperative cytology was satisfactory in 35 of 38 cases and positive in 17 (49%) after a mean of 6.9 months. Follow-up of these 35 transurethral resections disclosed a 15/17 (88%) recurrence rate after positive cytology and a 4/18 (22%) recurrence rate after negative cytology (P < .001). Postoperative cytology demonstrated a sensitivity for recurrence of 79%, specificity of 88%, positive predictive value of 88% and negative predictive value of 77%. In contrast, tumor in the transurethral resection specimen had a positive predictive value of 54% for recurrence, and its grade and stage were inferior to cytology in predicting recurrence. CONCLUSION: Postoperative bladder washing cytology is a useful adjunct to the management of papillary urothelial carcinoma. A positive result, signifying residual tumor, should encourage prompt follow-up and possibly repeat transurethral resection.     

FKBP38蛋白在乳腺癌中的表达研究

摘要 目的：探讨乳腺癌组织FK506结合蛋白38（FK506-binding protein 38,FKBP38）的表达水平及其与病理分级和临床分期的关系。方法：采用免疫组化检测100例正常乳腺组织、300例浸润性导管癌（Invasion ductal carcinoma,IDC）和59例浸润性小叶癌组织（Invasion lobular carcinoma,ILD）中FKBP38的表达水平,分析FKBP38蛋白表达水平与乳腺癌临床病理参数之间的关系。结果：免疫组化结果表明,与正常乳腺组织相比,FKBP38在浸润性导管癌及浸润性小叶癌组织中的表达水平均显著降低,具有统计学差异（P<0.0001）。通过进一步分析可知,在浸润性导管癌中,FKBP38蛋白表达水平随病理分级及临床分期的增加而降低,具有统计学差异,而FKBP38蛋白与孕激素受体（Progesterone receptor,PR）蛋白的表达呈负相关。此外,三阴性乳腺癌（Triple-negative breast cancer,TNBC）FKBP38的表达水平显著高于非三阴性乳腺癌,同样,FKBP38在TNBC的表达水平随原发性肿瘤分期的增加而降低。结论：FKBP38蛋白水平在乳腺癌患者中表达水平显著降低,并与乳腺癌的病理分级、临床分期相关。这提示FKBP38蛋白水平可作为乳腺癌诊断和治疗的潜在靶点,但其作用机制仍需进一步研究。     

Correlation between angiogenesis, apoptosis and cell proliferation in invasive ductal carcinoma of the breast and their relation to tumor behavior

OBJECTIVE: To investigate the relationship between angiogenesis, apoptosis and cell proliferation in invasive ductal carcinoma of the breast and their relation to tumor behavior. STUDY DESIGN: Microvessels were immunohistochemically labeled with antibody to CD34 in sections from 82 cases of invasive ductal carcinoma. Computerized image analysis was used to evaluate microvessel density (MVD). The authors measured the apoptotic index (AI) using the terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling technique and proliferating cell nuclear antigen labeling index (PCNA LI) by PCNA immunohistochemistry on serial sections. RESULTS: Statistical analysis revealed a significant inverse correlation between MVD and AI (r = -.313, P = .004) and failed to find a significant correlation between MVD and PCNA LI. There was a significant positive correlation between AI and PCNA LI (r = .393, P = .000). Significant differences in AI between high MVD (> or = 59.9%) and low MVD (< 59.9%) were seen (P < .001), with no appreciable differences in PCNA LI between the two groups. Histologic grade and stage were the only independent prognostic factors in both disease-free and overall survival. CONCLUSION: Angiogenesis in breast cancer may be related to the ability of tumor cells to survive rather than to their proliferative activity. Apoptosis is related to cell proliferation in breast cancer.     

nm23、EGFR和Bcl-2蛋白在肺癌组织中的表达及临床意义

目的:观察nm23、EGFR和Bcl-2在人肺癌中的表达,探讨其与肺癌临床生物学行为的关系。方法:采用免疫组化检测了49例肺癌及癌旁组织中nm23、EGFR和Bcl-2蛋白的表达情况,并对其与肺癌临床生物学行为进行相关分析。结果:在49例肺癌中,EGFR和Bel-2蛋白的阳性表达率分别为51.0%和34.7%,均高于正常肺组织17.5%和12.5%(p<0.05),nm23蛋白的阳性表达率为55.1%,则显著低于正常肺组织87.5%(p<0.01)。在病理分级的高一中分化、低分化和未分化中nm23的阳性表达率分别为82.6%、41.2%和11.1%(p<0.01)。EGFR在非小细胞肺癌(NSCLC)组为61.5%,在小细胞肺癌(SCLC)组为10.0%(p<0.01)。在NSCLC TNM分期中,Ⅰ期 Ⅱ期与Ⅲ P<0.01)期中EGFR的阳性表达率分别为39.3%和81.8%(p<0.01);病理分级的高-中分化和低分化中EGFR的阳性表达率分别为39.1%和88.2%(p<0.01);在淋巴结转移组为57.9%,无淋巴结转移组为27.3%(p<0.05)。Bcl-2的阳性表达率在NSCLC组为25.6%,在SCLC组为70%,统计学分析差异有统计学意义(p<0.01);在NSCLC组病理分级的高分化、中分化和低分化中Bcl-2阳性表达率分别为62.5%、26.7%和5.89%(p<0.01)。结论:癌基因EGFR、凋亡抑制基因Bcl-2与抑癌基因nm23在肺癌的发生、发展中存在着相互促进作用。     

信号转导与转录因子3和表皮生长因子受体在宫颈不同病变组织中的表达及其临床意义

目的:探讨信号转导与转录因子3(STAT3)与表皮生长因子受体(EGFR)在宫颈不同病变组织中的表达变化及其临床意义。方法:采用Western blot与免疫组化的方法检测100例上皮内瘤样病变(CIN)组织、60例宫颈癌组织及40例正常宫颈组织中STAT3与EGFR的表达;分析STAT3与EGFR的表达与宫颈癌组织病理特征的关系;对宫颈癌组织中STAT3与EGFR的表达的相关性进行分析。结果:宫颈癌组织以及CIN组织中STAT3、EGFR的表达量和阳性率显著高于正常宫颈组织(P0.05),且宫颈癌组织中STAT3、EGFR的表达量和阳性率高于CIN组织,差异有统计学意义(P0.05);STAT3与EGFR在宫颈癌组织中的染色强度较正常宫颈组织以及CIN组织显著增强;STAT3与EGFR的异常表达与宫颈癌患者的组织学分级、临床分期以及是否发生淋巴结转移有关(P0.05),而与年龄以及病理分型无关(P0.05);宫颈癌组织中STAT3和EGFR表达呈正相关(P0.05)。结论:STAT3与EGFR表达与CIN、宫颈癌的进展紧密相关,且与宫颈癌部分病理特征相关,二者有可能作为宫颈癌诊断及预后的参考指标。     

Manual versus image analysis estimation of PCNA in breast carcinoma

OBJECTIVE: To compare manual to image analysis estimation of proliferating cell nuclear antigen (PCNA) expression in paraffin sections of breast carcinomas. STUDY DESIGN: Paraffin sections of 51 breast carcinomas were stained with primary antibody to PCNA. Nuclear PCNA expression in 100 randomly selected tumor cells from marked areas was manually graded from 0 to 3. Antigen expression was also calculated by a cell analysis system (CAS-200, Becton Dickinson, Elmhurst, Illinois, U.S.A.) from marked and random microscopic fields. Obtained proliferative index (PI) from both methods was compared. RESULTS: Manually calculated PI correlated strongly with the CAS-200-calculated PI (P < .01). The highest correlation was seen between the CAS-200 PI value and manually calculated PI value using grade 2 and 3 nuclei. A particularly high correlation was noted between the number of positive nuclei and antigen staining area (P < .01) as estimated by the CAS-200. CONCLUSION: Nuclear expression of PCNA and other nuclear antigens can be accurately evaluated by an image analysis system. The speed and objectivity of such machines allow the evaluation of larger parts of tissues and provide more-representative antigen expression profiles.     

The assessment of multiple variables on breast carcinoma fine needle aspiration (FNA) cytology specimens: method, preliminary results and prognostic associations

The assessment of multiple variables on breast carcinoma fine needle aspiration (FNA) cytology specimens: method, preliminary results and prognostic associations
We have assessed multiple biological variables on breast carcinoma FNA specimens using a Cytoblock technique. The growth fraction (MIBI), oestrogen receptor (ER), progesterone receptor (PR), p53 mutant protein, c-erbB-2, epidermal growth factor receptor (EGFR), NCRCl Vepithelial membrane antigen (EMA) and DNA plopidy were examined. Objective quantification using image analysis (CAS 200) was applied as appropriate. Fifty cases were examined in this preliminary study. Excellent correlation between the Cytoblock preparations and parallel tissue sections was seen. Of the cancers, 81% were aneuploid with only 19% diploid in character, but 67% of the carcinomas were of histological grade 3. The mean nuclear area staining with MIBl was 31.3% and with ER was 26.7%. Twenty-four percent (24.1%) of the nuclear area showed immunoreactivity with PR. Significant immunostaining was seen in 38%, 46%, 38% and 95% of carcinomas with c-erbB-2, p53, EGFR and EMA, respectively. A significant association between histological grade of the resected tumours and both MIBl (P=0.04) and EGFR (P=0.02) expression in the Cytoblock samples was seen. p53 (P = 0.03) and EGFR (P=0.01) immunoreactivity showed an association with tumour size. EGFR (P=0.04) immunostaining also showed a relationship with the lymph node status of the patient. The technique is, we believe, a useful one for the assessment of multiple variables on breast cytology specimens; these preliminary data suggest that some of these may be useful in predicting prognosis in breast cancer patients.     

乳腺浸润性导管癌组织中AIB1 与Ki67的表达及临床意义

目的:探讨乳腺浸润性导管癌(IDC)中乳腺癌扩增性抗原1(AIB1)和增殖细胞核抗原(Ki67)蛋白的表达及临床意义。方法:选择2012年6月到2014年6月在我院经病理检查确诊为IDC患者的组织石蜡标本160例,采用链霉素-生物素(SP)免疫组化法检测标本中AIB1和Ki67蛋白的表达,多因素Logistic回归分析二者与IDC临床病理学特征的相关性。结果:AIB1、Ki67的阳性表达率分别为75.63%和80.63%。AIB1、Ki67的表达与淋巴结转移、组织学分级及TNM分期存在相关性(P0.05),且随组织学分级和TNM分期的增高,阳性表达率逐渐增高(P0.05),Ki67的表达水平随肿瘤变大,阳性率逐渐增加(P0.05)。淋巴结阳性组AIB1、Ki67的阳性表达率显著高于淋巴结阴性组(P0.05)。多因素Logistic回归分析显示,AIB1、Ki67的阳性表达是淋巴结转移、病理组织学分级及TNM分期的危险因素(P0.05)。结论:在IDC组织中AIB1和Ki67的阳性表达均增高,二者与IDC临床病理学特征有密切关系。     

CCEPR is a novel clinical biomarker for prognosis and regulates cell proliferation through PCNA in osteosarcoma

CCEPR (cervical carcinoma expressed PCNA regulatory lncRNA) has been found to be upregulated and enhance cell proliferation in human cancers. However, the role of CCEPR in osteosarcoma remains to be discovered. In this study, we found CCEPR expression was elevated in osteosarcoma tissue specimens and cell lines compared with adjacent normal tissue specimens and osteoblast cell line, respectively. Meanwhile, osteosarcoma patients with advanced stage or tumor size greater than 8 cm had higher expression of CCEPR than patients with early stage or tumor size less than or equal to 8 cm, respectively. Survival analysis suggested that osteosarcoma patients with high CCEPR expression had a worse overall survival rate than those with low CCEPR expression. The in vitro study indicated that CCEPR positively regulated proliferating cell nuclear antigen (PCNA) expression in osteosarcoma cells and silencing of CCEPR inhibited osteosarcoma cell proliferation through decreasing PCNA expression. In conclusion, CCEPR is a potential prognostic predictor and functions as oncogenic long non-coding RNA (lncRNA) to regulate cell proliferation via PCNA in osteosarcoma.     

Immunohistochemical detection of EGFR in paraffin-embedded tumor tissues: variation in staining intensity due to choice of fixative and storage time of tissue sections.

The epidermal growth factor receptor (EGFR) is highly expressed in a variety of solid malignant tumors and its expression has been correlated with disease progression and poor survival. With the advent of targeted therapies, especially IMC-C225 (Cetuximab), a monoclonal antibody (MAb) directed against the EGFR, there is an increasing interest in immunohistochemistry (IHC)-based EGFR screening methods using paraffin-embedded tumor specimens to select cancer patients eligible for treatment with Cetuximab. With the EGFRpharmDX kit, a complete assay for demonstration of EGFR is now available. Because no information about the preservation of the EGFR under various conditions of fixation is available, we performed a prospective study on a panel of commonly used fixatives to determine optimal tissue preservation protocols. The stability of the epitope on cut tissue sections stored for a period up to 24 month was also tested using material originating from patients with head and neck cancer, non-small-cell lung carcinomas, and colorectal adenocarcinomas. Depending on the fixative used and the time of storage of cut tissue sections, a variation in the determined level of EGFR expression was demonstrated compared with the most optimal fixation procedure.     

Fn14 在 EGFR 外显子 19 缺失突变的非小细胞肺癌中的表达及其临床意义

目的:探讨成纤维细胞生长因子诱导14(Fn14)在表皮生长因子受体(EGFR)外显子19缺失的非小细胞肺癌(NSCLC)组织中的表达及其临床意义。方法:选择2010年9月至2013年11月第四军医大学唐都医院收治的125例原发性EGFR外显子19缺失的NSCLC组织及与之相对应的30例正常肺组织为研究对象,应用免疫组化法检测和比较其Fn14的表达,分析Fn14的表达与原发性EGFR外显子19缺失的的NSCLC临床病理特征的关系。结果:Fn14阳性表达主要定位于胞膜和胞质中。在125例原发性EGFR外显子19缺失的NSCLC组织中,Fn14的阳性表达率为100%,显著高于正常肺组织(P<0.001)。鳞癌和腺癌NSCLC组织中Fn14的阳性表达率比较无统计学差异(P=0.106);不同病理分化程度、不同TNM分期的NSCLC组织中Fn14的阳性表达率比较均具有显著性差异(P=0.000、P=0.000)。Fn14蛋白的表达与EGFR外显子19缺失的NSCLC的淋巴结转移状态、肿瘤大小以及肿瘤解剖学分类均显著相关(P=0.000、P=0.029、P=0.000、P=0.026),而与患者的性别、年龄、吸烟史均无关(P=0.816、P=0.122、P=0.816)。结论:Fn14在原发性EGFR外显子19缺失的NSCLC中呈异常高表达,EGFR外显子19缺失突变很可能通过上调Fn14通路,促进NSCLC的发生和发展。     

FLIP、FADD在上皮性卵巢癌中表达及其临床意义

目的：研究FLIP、FADD在上皮性卵巢癌组织中的表达情况,探讨二者在上皮性卵巢癌发生发展中的作用及其临床意义。方法：采用免疫组化SP法及RT—PCR法检测44例上皮性卵巢癌及17例正常卵巢组织中FLIP、FADD的表达,并检测上皮性卵巢癌中PCNA的表达,分析FLIP,FADD表达与上皮性卵巢癌临床病理参数及PCNA表达的关系。结果：FLIP、FADD在上皮性卵巢癌组织中的阳性表达率,与正常卵巢组织相比,差异有统计学意义（P〈0．05）。FLIP,FADD表达与上皮性卵巢癌病理分级及临床分期有关,FLIP、FADD在I—II期中的阳性表达率,与III．IV期相比,差异有统计学意义（P〈0．05）。FLIP、FADD在病理组织学-1级中的阳性表达率,与2—3级相比,差异有统计学意义（P〈0．05）。FLIP、FADD的表达与患者年龄、组织学分型及淋巴结转移无关。上皮性卵巢癌组织中FLIP表达与PCNA表达成线性正相关,r分别0．880和0．564;FADD表达与PCNA表达成线性负相关,r分别为-0．591和-0．683。结论：FLIP、FADD与上皮性卵巢癌的发生发展密切相关,可能是治疗上皮性卵巢癌的潜在靶点。     

LncRNA TP73-AS1 predicts poor prognosis and functions as oncogenic lncRNA in osteosarcoma

TP73 antisense RNA 1 (TP73-AS1), a novel long noncoding RNA (lncRNA), has been suggested to be deregulated in various human cancers and serve as a tumor suppressor or promoter, depending on tumor types. The role of TP73-AS1 in osteosarcoma is still unknown. In our results, TP73-AS1 was highly expressed in osteosarcoma tissue samples and cell lines compared with matching adjacent nontumor tissue specimens and a normal human osteoblast cell line, respectively. Moreover, high expression of TP73-AS1 was statistically associated with advanced Enneking stage, large tumor size, present distant metastasis, and poor histological grade, while exhibiting no statistical association with age, sex, and tumor site. The survival analyses showed that patients with osteosarcoma with high expression of TP73-AS1 obviously had lower overall survival than osteosarcoma patients with low expression of TP73-AS1, and high expression of TP73-AS1 was an independent poor prognostic factor for osteosarcoma patients. The experiments in vitro indicated that inhibition of TP73-AS1 expression depressed osteosarcoma cell viability, migration, and invasion, and arrested cell cycle. In conclusion, TP73-AS1 serves as oncogenic lncRNA participated in osteosarcoma progression.     

Overexpression of RBBP6, Alone or Combined with Mutant TP53, Is Predictive of Poor Prognosis in Colon Cancer

Retinoblastoma binding protein 6 (RBBP6) plays an important role in chaperone-mediated ubiquitination and interacts with TP53 in carcinogenesis. However, the clinicopathologic significance of RBBP6 expression in colon cancer is unknown; in particular, the prognostic value of RBBP6 combined with TP53 expression has not been explored. Therefore, quantitative real-time PCR and western blot analyses were performed to detect RBBP6 expression in colon cancer tissues. RBBP6 and TP53 expression were assessed by immunohistochemistry in a tissue microarray format, in which the primary colon cancer tissue was paired with noncancerous tissue. Tissue specimens were obtained from 203 patients. We found that RBBP6 was overexpressed in colon tumorous tissues and was significantly associated with clinical stage, depth of tumor invasion, lymph node metastasis (LNM), distant metastasis, and histologic grade. Further studies revealed that a corresponding correlation between RBBP6 overexpression and mutant TP53 was evident in colon cancer (r = 0.450; P<0.001). RBBP6 expression was an independent prognostic factor for overall survival (OS) and disease free survival (DFS). Interestingly, patients with tumors that had both RBBP6 overexpression and mutant TP53 protein accumulation relapsed and died within a significantly short period after surgery (P<0.001). Multivariate analysis showed that patients with LNM and patients with both RBBP6- and TP53-positive tumors had extremely poor OS (HR 6.75; 95% CI 2.63–17.35; P<0.001) and DFS (HR 8.08; 95% CI 2.80–23.30; P<0.001). These clinical findings indicate that the assessment of both RBBP6 and mutant TP53 expression will be helpful in predicting colon cancer prognosis.     

EGFR soluble isoforms and their transcripts are expressed in meningiomas

The EGFR (epidermal growth factor receptor) is involved in the oncogenesis of many tumors. In addition to the full-length EGFR (isoform a), normal and tumor cells produce soluble EGFR isoforms (sEGFR) that lack the intracellular domain. sEGFR isoforms b, c and d are encoded by EGFR variants 2 (v2), 3 (v3) and 4 (v4) mRNA resulting from gene alternative splicing. Accordingly, the results of EGFR protein expression analysis depend on the domain targeted by the antibodies. In meningiomas, EGFR expression investigations mainly focused on EGFR isoform a. sEGFR and EGFRvIII mutant, that encodes a constitutively active truncated receptor, have not been studied. In a 69 meningiomas series, protein expression was analyzed by immunohistochemistry using extracellular domain targeted antibody (ECD-Ab) and intracellular domain targeted antibody (ICD-Ab). EGFRv1 to v4 and EGFRvIII mRNAs were quantified by RT-PCR and EGFR amplification revealed by MLPA. Results were analyzed with respect to clinical data, tumor resection (Simpson grade), histological type, tumor grade, and patient outcome.Immunochemical staining was stronger with ECD-Ab than with ICD-Ab. Meningiomas expressed EGFRv1 to -v4 mRNAs but not EGFRvIII mutant. Intermediate or high ECD-Ab staining and high EGFRv1 to v4 mRNA levels were associated to a better progression free survival (PFS). PFS was also improved in women, when tumor resection was evaluated as Simpson 1 or 2, in grade I vs. grade II and III meningiomas and when Ki67 labeling index was lower than 10%. Our results suggest that, EGFR protein isoforms without ICD and their corresponding mRNA variants are expressed in meningiomas in addition to the whole isoform a. EGFRvIII was not expressed. High expression levels seem to be related to a better prognosis. These results indicate that the oncogenetic mechanisms involving the EGFR pathway in meningiomas could be different from other tumor types.     

Intratumor heterogeneity of biomarker expression in breast carcinomas

Small biopsy samples are used increasingly to assess the biomarker expression for prognostic information and for monitoring therapeutic responses prior to and during neoadjuvant therapy. The issue of intratumor heterogeneity of expression of biomarkers, however, has raised questions about the validity of the assessment of biomarker expression based on limited tissue samples. We examined immunohistochemically the expression of HER-2neu (p185^erbB-2), epidermal growth factor receptor (EGFR), Bcl-2, p53, and proliferating cell nuclear antigen (PCNA) in 30 breast carcinomas using archived, paraffin embedded tissue and determined the extent of intratumor heterogeneity. Each section was divided into four randomly oriented discrete regions, each containing a portion of the infiltrating carcinoma. For each tumor, the entire lesion and four regions were analyzed for the expression of these markers. Scores of both membrane and cytoplasmic staining of HER-2neu and EGFR, scores of cytoplasmic staining of Bcl-2, and scores of nuclear staining of both p53 and PCNA were recorded. The intensity of staining and the proportion of immunostained cells were determined. A semiquantitative immunoscore was calculated by determining the sum of the products of the intensity and corresponding proportion of stained tumor cells. We analyzed both invasive (IDC) and in situ (DCIS) carcinomas. The Wilcoxon signed-rank test was used for paired comparisons between overall and regional immunoscores and between overall and regional percentages of stained cells. Spearman's correlation coefficients were used to assess the level of agreement of overall biomarker expression with each of the regions. Generalized linear models were used to assess overall and pair-wise differences in the absolute values of percent changes between overall and regional expression of biomarkers. For IDCs, there were no statistically significant differences in the expression of the biomarkers in terms of either the percentage of cells staining or the immunoscores when comparing the entire tumor with each region except for the lower EGFR expression of arbitrarily selected region 1 and lower p53 expression of region 1 compared to that of the entire tumor section. For DCIS, there were no statistically significant differences in the expression of the biomarkers between the entire tumor and each region except in PCNA of region 2 compared to that of entire tumor section. Positive correlation of immunoscores was observed between the entire tumor and each region as well as across all four regions for IDC. Similar observations were noted with DCIS except for HER-2neu and PCNA. No statistically significant differences were observed in the absolute values of percent changes of biomarker expression between overall and the four regions for both DCIS and IDC. Therefore, no significant intratumor heterogeneity in the expression of HER-2neu, Bcl-2, and PCNA was observed in IDC. Minor regional variations were observed for EGFR and p53 in IDC. Similarly, no significant regional variation in the expression of markers was observed in DCIS except for PCNA.     

The prognostic significance of thymidine phosphorylase, thymidylate synthase and dihydropyrimidine dehydrogenase mRNA expressions in breast carcinomas

Thymidine phosphorylase (TP), thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) have been indicated as possible predictive markers for epithelial malignancies. All these three enzymes are actively involved in 5-FU metabolism. In this report, we investigated mRNA expression of these factors with real-time quantitative PCR in a series of 86 micro-selected breast carcinomas and 8 micro-selected tumour-adjacent normal breast epithelial specimens. Highly variable mRNA expressions of these factors were observed in both normal and cancerous samples. TP and TS mRNA expressions in breast carcinomas were elevated, but only TS mRNA expression showed a trend for statistical difference, compared with the expression in normal breast epithelial samples. Although the DPD mRNA expression range in tumours was also elevated, the average mean was reduced in tumours compared to that in normal samples. No association between mRNA expressions of TP, TS and DPD and clinicopathological features such as histological grade, tumour size, node status, S-phase fraction, ploidy, and clinical stage was found. A negative association between DPD mRNA expression and age was, however, revealed. Ten-year follow-up analysis showed no association between TP and DPD mRNA expression and clinical outcome. An high level of TS mRNA expression, however, was associated with a shorter clinical survival, indicating its potential role as a clinical marker in breast carcinoma.     

Prognostic significance of p53, bcl-2 and EGFR in carcinoma of the endometrium

OBJECTIVE: To evaluate the part played by several parameters in the prognosis of patients with endometrial carcinoma. STUDY DESIGN: Eighty imprint smears from fresh endometrial tumor specimens were studied immunocytochemically for the expression of p53, bcl-2 and epidermal growth factor receptor. Also, the presence of estrogen receptor (ER) and progesterone receptor (PR) in the tumor tissue was measured. The data obtained were related to survival, and associations were sought between the parameters studied. RESULTS: Strong associations were found between advanced stage, high grade, lymph node metastases at diagnosis, nonendometrioid histology and p53 expression with poor survival. Bcl-2 expression was associated with good five-year survival. ER expression was associated marginally with good five-year survival, but PR expression was not. A strong association was found between p53 and advanced disease, stage and lymph node metastases at diagnosis. An association between EGFR positivity and survival was not found. CONCLUSION: p53 Expression of uterine tumors is an independent and strong indicator of poor prognosis. Even patients with stage I and II disease at surgery who have p53-positive tumors must be considered at high risk.