ASPM and microcephalin expression in epithelial ovarian cancer correlates with tumour grade and survival

Background:

The clinico-pathological and molecular heterogeneity of epithelial ovarian cancer (EOC) complicates its early diagnosis and successful treatment. Highly aneuploid tumours and the presence of ascitic fluids are hallmarks of EOC. Two microcephaly-associated proteins, abnormal spindle-like microcephaly-associated protein (ASPM) and microcephalin, are involved in mitosis and DNA damage repair. Their expression is deregulated at the RNA level in EOC. Here, ASPM and microcephalin protein expression in primary cultures established from the ascites of patients with EOC was determined and correlated with clinical data to assess their suitability as biomarkers.

Methods:

Five established ovarian cancer cell lines, cells derived from two benign ovarian ascites samples and 40 primary cultures of EOC derived from ovarian ascites samples were analysed by protein slot blotting and/or immunofluorescence to determine ASPM and microcephalin protein levels and their cellular localisation. Results were correlated with clinico-pathological data.

Results:

A statistically significant correlation was identified for ASPM localisation and tumour grade, with high levels of cytoplasmic ASPM correlating with grade 1 tumours. Conversely, cytoplasmic microcephalin was only identified in high-grade tumours. Furthermore, low levels of nuclear microcephalin correlated with reduced patient survival.

Conclusion:

Our results suggest that ASPM and microcephalin have the potential to be biomarkers in ovarian cancer.     

Gene expression analysis using long-term preserved formalin-fixed and paraffin-embedded tissue of non-small cell lung cancer

This study was performed to evaluate RNA extraction and gene expression analysis of non-small cell lung cancer (NSCLC) using formalin-fixed and paraffin-embedded (FFPE) specimens stored for more than 20 years by quantitative PCR (qPCR) and DNA microarrays. Long-term preserved FFPE materials enable large retrospective studies correlating molecular features with therapeutic response and clinical outcome. qPCR was used to evaluate RNA extraction methods and to compare DNA microarray gene expression profiles of FFPE and fresh frozen (FF) tissue. The Ambion RecoverAll kit appeared to be suited for RNA extraction of long-term preserved FFPE tissues. Microarray analysis using the Affymetrix platform displayed a high degree of correlation for endogenous control genes comparing FF and FFPE tissues and identified known NSCLC signature genes in both specimens. We conclude that high quality gene expression signatures can be recognized using the Affymetrix gene expression platform on FFPE tissue stored for more than 20 years. However, a general interpretation must be done with caution as different FFPE procedures have varying effects on RNA quality.     

石蜡包埋的肝细胞癌组织中ASPP家族的表达

目的:研究肝细胞癌中p53凋亡刺激蛋白（ASPP）家族的表达以及临床意义。方法:应用免疫组织化学SP方法检测73例甲醛固定和石蜡包埋的肝细胞癌组织切片和相应癌旁正常组织中ASPP蛋白家族的表达情况。结果:在癌旁正常组织中,ASPP1和ASPP2的表达阳性率高于肝细胞癌组织(P=0.000,0.000),但是肝细胞癌组织中iASPP的表达阳性率高于相应癌旁正常组织(P=0.000)。在肝细胞癌组织中,ASPP1的表达与患者性别、年龄、临床分期以及淋巴结转移均无相关性(P＞0.05),但与病理分级有关(P=0.001);ASPP2的表达与病理分级和临床分期有关(P=0.000,0.000),与性别、年龄以及淋巴结转移无关(P＞0.05)。iASPP的表达与淋巴结转移有关(P=0.004),但是与性别、年龄、病理分级和临床分期无关(P＞0.05)。结论:ASPP蛋白家族异常表达可能与肝细胞癌的发生机制相关,ASPP1和ASPP2表达水平下调并与肝细胞癌病理分级相关,而 iASPP的表达水平上调与癌的淋巴转移相关,iASPP可能是基因治疗肝细胞癌的一个靶点。      

Direct cancer tissue proteomics: a method to identify candidate cancer biomarkers from formalin-fixed paraffin-embedded archival tissues

Successful treatment of multiple cancer types requires early detection and identification of reliable biomarkers present in specific cancer tissues. To test the feasibility of identifying proteins from archival cancer tissues, we have developed a methodology, termed direct tissue proteomics (DTP), which can be used to identify proteins directly from formalin-fixed paraffin-embedded prostate cancer tissue samples. Using minute prostate biopsy sections, we demonstrate the identification of 428 prostate-expressed proteins using the shotgun method. Because the DTP method is not quantitative, we employed the absolute quantification method and demonstrate picogram level quantification of prostate-specific antigen. In depth bioinformatics analysis of these expressed proteins affords the categorization of metabolic pathways that may be important for distinct stages of prostate carcinogenesis. Furthermore, we validate Wnt-3 as an upregulated protein in cancerous prostate cells by immunohistochemistry. We propose that this general strategy provides a roadmap for successful identification of critical molecular targets of multiple cancer types.     

Correlation between sigma2 receptor protein expression and histopathologic grade in human bladder cancer

Sigma2 (sigma2) receptor proteins are overexpressed in several tissues and tumour cell lines. Although the biomolecular mechanism of this overexpression must be elucidated, sigma2 receptor was considered a potential biomarker for monitoring solid tumour proliferation. In this study, we verified first sigma2 receptor overexpression by saturation analysis with radioligand in six human bladder cancer specimens, then if a possible correlation could be established between sigma2 overexpression and tumour tissue stage and grade. The results displayed that sigma2 receptor protein was normally expressed in human bladder and overexpressed in the case of high-grade transitional cell carcinomas. Moreover, these receptors were undetected in a low-grade squamous cell carcinoma and in a very rare form of anaplastic, large cells plasmacytoid cancer.     

钙离子结合蛋白A4和上皮型钙黏附蛋白在膀胱癌中的表达及临床意义

目的：探讨钙离子结合蛋白A4（S100A4）及上皮性钙黏附蛋白（E—cadherin）在膀胱癌中的表达和临床意义。方法：应用免疫组化S—P法检测64例膀胱移行细胞癌组织及10例正常人膀胱组织中S100A4蛋白及E—cad,herin的表达。结果：正常膀胱组织S100A4蛋白表达阴性，E—cadherin表达阳性。64例膀胱移行细胞癌组织中46例（71．9％）s100A4蛋白阳性表达，36例（56．3％）E—cadherin蛋白阳性表达。正常对照膀胱组织与膀胱移行细胞癌组织之间S100A4蛋白及E—cadherin表达差异有统计学意义（均为P〈0．05）S100A4蛋白及E—cadherin在肿瘤组织中表达明显增高，与膀胱移行细胞癌的组织学分级、临床（T）分期和淋巴结转移有关，而与年龄、性别和肿瘤大小无关。结论：在膀胱移行细胞癌中，S100A4蛋白及E—cadherin的表达变化与肿瘤的进展密切相关，是判断其生物学行为，预测转移趋势极具价值的指标。     

The relationship between T-lymphocyte infiltration, stage, tumour grade and survival in patients undergoing curative surgery for renal cell cancer

The present study examined the relationship between tumour stage, grade, T-lymphocyte subset infiltration and survival in patients who had undergone potentially curative surgery for renal clear-cell cancer (n=73). Intratumoural CD4+ T-lymphocyte infiltrate was associated with poor cancer-specific survival, independent of grade, in this cohort.     

Gene expression patterns in formalin-fixed, paraffin-embedded core biopsies predict docetaxel chemosensitivity in breast cancer patients

Previously, we had identified gene expression patterns that predicted response to neoadjuvant docetaxel. Other studies have validated that a high Recurrence Score (RS) by the 21-gene RT-PCR assay is predictive of worse prognosis but better response to chemotherapy. We investigated whether tumor expression of these 21 genes and other candidate genes can predict response to docetaxel. Core biopsies from 97 patients were obtained before treatment with neoadjuvant docetaxel (4 cycles, 100 mg/m² q3 weeks). Three 10-μm FFPE sections were submitted for quantitative RT-PCR assays of 192 genes that were selected from our previous work and the literature. Of the 97 patients, 81 (84%) had sufficient invasive cancer, 80 (82%) had sufficient RNA for QRTPCR assay, and 72 (74%) had clinical response data. Mean age was 48.5 years, and the median tumor size was 6 cm. Clinical complete responses (CR) were observed in 12 (17%), partial responses in 41 (57%), stable disease in 17 (24%), and progressive disease in 2 patients (3%). A significant relationship (P < 0.05) between gene expression and CR was observed for 14 genes, including CYBA. CR was associated with lower expression of the ER gene group and higher expression of the proliferation gene group from the 21 gene assay. Of note, CR was more likely with a high RS (P = 0.008). We have established molecular profiles of sensitivity to docetaxel. RT-PCR technology provides a potential platform for a predictive test of docetaxel chemosensitivity using small amounts of routinely processed material.     

Proteome profiling of formalin-fixed,paraffin-embedded lung adenocarcinoma tissues using a tandem mass tag-based quantitative proteomics approach

Over the past few decades, increasing efforts have been made to improve the understanding of, and treatment options for, lung adenocarcinoma (LUAD). However, considering the heterogeneity of LUAD, precise proteomics-based characterization at the molecular level is an urgent clinical requirement for effective treatment. Formalin-fixed, paraffin-embedded (FFPE) tissue is a good option as the working tool for proteomics studies. The present study aimed to obtain a global protein profile using LUAD FFPE tissue samples. Using a quantitative proteomics approach, the study revealed that 360 proteins were significantly more highly expressed in LUAD than in adjacent nontumor lung tissues. Also, 19 differentially expressed membrane proteins were found to be primarily responsible for immune processes. Epidermal growth factor (EGF)-like domain and laminin EGF domain showed markedly different expression levels between cancer tissues and tumor-adjacent normal tissues. Furthermore, Gene Ontology functional enrichment analysis showed that significantly upregulated proteins were associated with the endoplasmic reticulum lumen, protein disulfide isomerase activity, vitamin binding, cell cycle G₁/S phase transition, to name but a few. Also, numerous kinases and post-translational modification enzymes were significantly upregulated across all eight LUAD samples compared with paracarcinoma tissues. Proteomics analysis revealed that AAA domain containing 3A (ATAD3a), a member of the ATPase family, was highly expressed in LUAD tissues, which was supported by immunohistochemical analysis. Furthermore, the study confirmed that ATAD3a enhanced the cisplatin sensitivity of LUAD cells. Collectively, the findings of the present study provide new potential candidate targets in patients with LUAD, and may aid auxiliary LUAD diagnosis and surveillance in a noninvasive manner.     

Molecular subtypes, histopathological grade and survival in a historic cohort of breast cancer patients

Molecular subtyping of breast cancer may provide additional prognostic information regarding patient outcome. However, its clinical significance remains to be established. In this study, the main aims were to discover whether reclassification of breast cancer into molecular subtypes provides more precise information regarding outcome compared to conventional histopathological grading and to study breast cancer-specific survival in the different molecular subtypes. Cases of breast cancer occurring in a cohort of women born between 1886 and 1928 with long-term follow-up were included in the study. Tissue microarrays were constructed from archival formalin-fixed, paraffin-embedded tissue from 909 cases. Using immunohistochemistry and in situ hybridisation as surrogates for gene expression analyses, all cases were reclassified into the following molecular subtypes: Luminal A; Luminal B (HER2?); Luminal B (HER2+); HER2 subtype; Basal phenotype; and five negative phenotype. Kaplan–Meier survival curves and Cox proportional hazards models were used in the analyses. During the first 5 years after diagnosis, there were significant differences in prognosis according to molecular subtypes with the best survival for the Luminal A subtype and the worst for HER2 and five negative phenotype. In this historic cohort of women with breast cancer, differences in breast cancer-specific survival according to subtype occur almost exclusively amongst the histopathological grade 2 tumours. From 5 years after time of diagnosis until the end of follow-up, there appears to be no difference in survival according to molecular subtype or histopathological grade.     

广泛期小细胞肺癌一线化疗肿瘤缓解深度与生存期的相关性

目的:分析广泛期小细胞肺癌一线化疗后肿瘤缓解深度与患者生存期的相关性。方法:回顾性分析符合入组条件的50例初治广泛期小细胞肺癌患者的临床资料。通过Spearman秩相关检验评价广泛期小细胞肺癌化疗后肿瘤缓解深度与生存期的相关性，应用Log-rank检验比较不同肿瘤缓解深度对生存期的影响，应用COX比例回归模型进行多因素分析。结果:Spearman秩相关分析显示肿瘤缓解深度与PFS及OS均呈中等程度相关。不同缓解深度患者的生存期存在统计学差异。体重减少（P＜0.000 1）、缓解深度（P＜0.001）为无进展生存期的独立影响因素；体重减少（P＜0.000 1）、体力状态（P=0.001 2）、缓解深度（P＜0.001）、化疗周期（P=0.000 2）、二线治疗（P=0.006 7）为总生存期的独立预后因素。结论:广泛期小细胞肺癌一线化疗后肿瘤缓解深度对患者生存期有一定的预测价值。