Bcl-2和nm23的表达与乳腺癌生物学行为的关系

目的研究乳腺癌中bcl-2与nm23基因表达的生物学意义及它们之间的相互关系对乳腺癌生物学行为的影响.方法应用免疫组织化学S-P方法对65例乳腺癌石蜡包埋组织进行检测,分析bcl-2与nm23基因表达与乳腺癌病理参数的关系.结果 bcl-2和nm23阳性表达率分别为57.4%、63.1%.bcl-2基因阳性表达与组织学类型、患者年龄、有无淋巴结转移以及肿瘤大小无关(P>0.05);与乳腺癌组织学分级呈正相关(P<0.05),nm23阳性表达与肿瘤大小、组织学类型和患者年龄无关(P>0.05);与乳腺肿瘤的组织学分级和淋巴结转移呈负相关(P<0.05);bcl-2与nm23的表达有相关性(P<0.05).结论 bcl-2阳性表达和nm23阴性的乳腺癌患者细胞恶性程度高,bcl-2与nm23基因共同参与了乳腺癌的发生和发展.     

乳腺癌外周血单个核细胞与间质免疫微环境的关系研究

目的探讨乳腺癌免疫微环境与外周血的关系。方法应用BRB-Array Tools软件对公共基因芯片数据库GEO中的乳腺癌间质及乳腺癌患者外周血单个核细胞基因芯片表达数据进行统计学分析,找出在乳腺癌间质及外周血单个核细胞均发生变化的基因,DAVID工具进一步分析其功能及参与的生物学通路。PINA蛋白质互作平台分析这些基因的蛋白质相互作用情况。结果比较后得到共同差异表达的103条基因,失调方向一致的基因70条,功能涉及炎症反应、髓系细胞分化、白细胞激活、抗原加工提呈等多种免疫相关的生物学过程。结论乳腺癌患者外周血单个核细胞基因表达改变,与肿瘤间质微环境具有一定相似度,有望建立基于外周血的免疫微环境分子预测,为乳腺癌的治疗靶点及预后判断的研究开辟新思路。     

鼻咽癌EMT相关基因的筛选及生物信息学分析

目的利用生物信息学方法挖掘鼻咽癌上皮间充质转化（epithelial-mesenchymal transition,EMT）发生的潜在分子机制。方法从公共基因芯片数据库GEO（gene expression omnibus）中寻找并下载鼻咽癌的相关基因芯片数据,并使用Genclip软件对下载的数据进行分析,寻找鼻咽癌公共基因芯片数据中与EMT相关的基因,并用生物信息学方法对筛选出来的基因作进一步分析。结果在公共的鼻咽癌芯片数据中找到35个与EMT相关的差异表达基因,这些基因功能大致与细胞组分、细胞粘附、通信、信号转导、分化、运动、迁移以及细胞表面受体相关的信号转导等有关,这些功能往往都被认为与肿瘤的侵袭和转移有关。结论利用生物信息学的方法能有效分析基因芯片数据并获取生物内在信息。鼻咽癌EMT是由于多种基因表达改变所致,这为确定鼻咽癌早期转移诊断标志与预后的预示开辟了新的思路。     

乳腺癌中miR-134的表达及其与上皮-间质转化的关系

目的：探讨miR-134在乳腺癌中的表达及临床意义,分析其表达与上皮-间质转化（ epithe1ia1-mesenchyma1 transition, EMT）标志物的相关性。方法采用原位分子杂交法检测97例乳腺癌组织中miR-134的表达,同时应用免疫组化MaxVision两步法检测E-cadherin、N-cadherin、vimentin的表达,并分析三者表达与临床病理特征的关系。结果 miR-134在乳腺癌中的阳性率随组织学分级的升高而降低,其表达与HER-2表达呈负相关。miR-134的阳性率随E-cadherin表达下调及N-cadherin、vimentin表达上调而降低;E-cadherin在乳腺癌中表达下调以及N-cadherin、vimentin在乳腺癌中表达上调均与组织学分级升高、淋巴结转移和HER-2阳性以及ER、PR阴性有关。结论 miR-134在乳腺癌组织中的表达随肿瘤恶性程度的升高而下调,并与EMT关系密切,是乳腺癌的潜在生物学标志物。     

三阴型乳腺癌中CD151的表达及意义

目的观察CD151在三阴型乳腺癌及非特殊型浸润性乳腺癌中的表达及与乳腺癌临床病理特征的关系。方法采用免疫组化EnVision两步法检测CD151在三阴型乳腺癌及非特殊型浸润性乳腺癌中的表达,运用统计学方法分析CD151表达与乳腺癌临床病理特征、无瘤生存期和预后的关系。结果在三阴型乳腺癌中,CD151表达与淋巴结转移和组织学分级相关(P0.05),CD151表达随着组织学分级的增加而升高,与患者年龄(P=0.081)和肿瘤大小(P=0.170)无明显相关性(P均0.05);三阴型乳腺癌患者肿瘤直径与腋窝淋巴结转移具有相关性(P0.05),CD151高表达患者的腋窝淋巴结转移率较高,无瘤生存期较短。结论 CD151在三阴型乳腺癌患者中高表达,并与淋巴结转移及组织学分级具有相关性,检测CD151有望成为评估三阴型乳腺癌患者预后的重要指标,为临床治疗和研究提供靶标。     

SASH1和p-ERK在乳腺浸润性导管癌中的表达及意义

目的观察SASH1和p-ERK在乳腺浸润性导管癌组织和正常乳腺组织中的表达,探讨其与乳腺浸润性导管癌生物学行为和预后的关系。方法应用免疫组织化学方法检测100例乳腺浸润性导管癌及40例正常乳腺组织中SASH1和p-ERK的表达,并结合肿瘤临床病理特征进行分析。Western blot方法检测20例新鲜乳腺浸润性导管癌组织及10例新鲜正常乳腺组织中SASH1和p-ERK的表达水平;实时荧光定量PCR方法检测两组SASH1mRNA和p-ERK mRNA的表达水平。结果 SASH1蛋白在乳腺癌组织表达率(31%)低于在正常乳腺组织表达率(80%),表达与组织学分级、淋巴结转移有关,而与肿瘤大小、TNM分期、发病年龄、激素受体表达无明显相关性。p-ERK蛋白在乳腺癌组织表达率(62%)高于正常乳腺组织(20%),表达与肿瘤大小、组织学分级呈正相关,与发病年龄、TNM分期、组织学分级、有无淋巴结转移、激素受体表达无明显相关。乳腺癌中SASH1的表达与p-ERK表达呈负相关(P0.005)。Western blot与实时PCR结果显示,与正常乳腺组织相比,SASH1蛋白与mRNA在乳腺癌组织中表达水平显著降低,p-ERK蛋白与mRNA在乳腺癌组织中表达水平显著升高(P0.01)。结论 SASH1的低表达和p-ERK的高表达与乳腺浸润性导管癌的发生发展密切相关。     

乳腺癌的分子分型及其临床意义

个体化治疗将成为乳腺癌治疗的发展方向,以生物学差异为划分依据的乳腺癌分子分型对个体化治疗方案的选择具有重要的参考价值.乳腺癌分子分型的研究已从高分子水平深入到基因水平,其相关因子包括ER、PR、HER2基因、p53基因、Ki-67蛋白表达等,这些生物学指标不但与肿瘤的生长、浸润、转移、复发等密切相关,而且对乳腺癌的治疗有指导性的研究价值.     

不同分化程度卵巢癌细胞中胚胎干细胞相关通路的基因集富集分析

背景:近年来一些研究发现肿瘤细胞具有与干细胞类似的特征,因此,控制干细胞功能的某些基因调控网络,可能也在某些肿瘤中同样发挥作用。目的:观察不同分化程度卵巢癌的分子特征,获得与胚胎干细胞相关基因的表达情况。方法:从美国国立生物信息中心(NCBI)共享数据库GEO下载原始基因芯片文件,登录号为GSE2109,选取卵巢癌样本作为研究材料。去除临床指标缺失的样本,按照组织学分级将卵巢癌样本分为高分化和低分化两组。原始数据经dChip进行质量检验和标准化,获得基因表达的矩阵。收集胚胎干细胞相关的基因集、生物学过程和KEGG通路在不同分化的卵巢癌中的富集情况,用GSEA进行基因集富集分析。结果与结论:选取了13个与胚胎干细胞相关的基因集,低分化的卵巢癌细胞中有9个基因集上调,PRC2靶点相关的4个基因集下调,说明与胚胎干细胞相关的基因集大多在低分化的卵巢癌细胞中上调。并且在低分化卵巢癌细胞显著上调的都是与细胞周期、细胞分裂、DNA复制等与增殖相关的通路。基因表达谱分析表明低分化的卵巢癌与胚胎干细胞具有相似的分子特征,这可以为卵巢癌的早期诊断和治疗提供新的思路。     

P53基因蛋白在乳腺良性病变和癌细胞表达的定量研究

应用流式细胞术和细胞免疫荧光染色技术，对乳腺良性病变和乳腺癌细胞Ｐ５３基因蛋白表达进行定量研究。结果表明，乳腺良性病变Ｐ５３蛋白表达量明显低于乳腺癌。Ｐ５３表达阳性率和表达量与乳腺癌的组织学分级密切相关，Ｐ５３表达量与乳腺癌ＤＮＡ倍体相关，非整倍体肿瘤Ｐ５３表达量高于二倍体肿瘤。Ｐ５３表达阳性（ＦＩ＞１．０）的乳腺癌，其生存期和五年生存率明显低于Ｐ５３表达阴性（ＦＩ≤１．０）的乳腺癌。     

基因芯片技术筛选主动脉瘤与正常主动脉差异表达基因

目的利用基因芯片技术研究人主动脉瘤组织与正常主动脉组织基因表达谱的差异。方法选取主动脉瘤病例标本5例,正常主动脉标本4例。提取总RNA,反转录成c DNA、体外转录合成aRNA后与芯片进行杂交,对结果进行分析。同时利用RT-qPCR对从表达谱筛选出的其中6个差异基因进行基因转录水平的定量验证。结果人主动脉瘤组与正常主动脉组相比,表达差异倍数大于2的基因共有270个,其中有211个基因上调表达,59个基因下调表达。差异表达的基因主要参与信号传导、免疫反应和炎性反应等生物学过程。RT-qPCR检测结果与基因芯片结果的符合率为100%。结论主动脉瘤与正常主动脉的表达谱有较大差异,利用基因芯片技术结合RT-qPCR验证可以筛选出主动脉瘤差异表达的基因,为研究主动脉瘤发病机制提供分子基础。     

Cytohistological correlation of grading in breast carcinoma

Assigning a tumor grade to breast cancer provides important prognostic information and guides optimal therapy. The present study was undertaken to grade breast carcinoma on aspirates by Robinson grading (RGS) and Scarff-Bloom-Richardson grading system (SBR). Histological grading was done according to Nottingham modification of Scarff-Bloom-Richardson method and the two cytological grading systems were compared. Fifty cases of breast carcinoma with preoperative cytologic diagnosis were assigned tumor grade. Histologic grading was done on corresponding mastectomy or partially resected specimens. Statistical analysis was performed; correlation between cytologic and histologic grading was established using Spearman correlation coefficient. Regression analysis was done to assess the significance of each cytological feature. Univariate analysis showed strong correlation (P < 0.01) for all features except dissociation. Multiple regression analysis of cytologic features revealed cell dissociation nucleoli and nuclear margin as the most influential features. A concordance of 72.5% between RGS (cyto) and SBR (cyto), 64% for RGS (cyto) and SBR (histo), 82% for SBR (cyto) and SBR (histo) was noted. There was a significant association (P < 0.001) between the grades assigned to cytologic and histologic specimens. Cytologic grade could be used to predict histologic grade as significant relationship exists between grades assigned to cytologic and histologic specimens.     

Reduced expression of APC and DCC gene protein in breast cancer

AIMS: To investigate if the adenomatous polyposis coli (APC) protein and the deleted in colorectal cancer (DCC) protein expression can be demonstrated by an immuno-histochemical method and to study the role of APC and DCC gene inactivation in the development and progression of breast cancer using colorectal cancer as a control model. METHODS AND RESULTS: The reduced or loss of protein expression of the APC and DCC genes was studied in 27 surgical specimens of primary breast cancer using an immunohistochemical method. Reduced or lost expression was identified in 11 out of 27 samples (40.7%) for the APC gene and 15 out of 27 samples (55.6%) for the DCC gene. No statistically significant difference was observed between the reduced or lost protein expression and the histological grading of breast tumour for both the APC and the DCC gene. CONCLUSIONS: Both gene proteins can be demonstrated by the immunohistochemical method. Reduced or loss of APC and DCC gene product were observed in 40.7% and 55.6% cases of primary breast cancer respectively. Further work is required to investigate the significance of the finding.     

In silico identification of breast cancer genes by combined multiple high throughput analyses

Publicly available human genomic sequence data provide an unprecedented opportunity for researchers to decode the functionality of human genome. Such information is extremely valuable in cancer prevention diagnosis and treatment. Cancer Genome Anatomy Project (CGAP) and Gene Expression Omnibus (GEO) are two bioinformatic infrastructures for studying functional genomics. The goal of this study is to explore the feasibility of incorporating the Internet-available bioinformatic databases to discover human breast cancer-related genes. Several tools including the Gene Finder, Virtual Northern (vNorthern) and SAGE digital gene expression displayer (DGED) were used to analyze differential gene expression between benign and malignant breast tissues. A pilot study was performed using both EST and SAGE vNorthern to analyze the expression of a panel of known genes, including high abundance genes beta-actin and G3PDH, low abundance genes BRCA1 and p53, tissue-specific genes CEA and PSA and two breast cancer-related genes Her2/neu and MUC1. We found a high expression of beta-actin and G3PDH and a low expression of BRCA1 and p53 across different types of tissues as well as a tissue-specific expression of CEA in colon and PSA in prostate. A further analysis of 30 known breast cancer-related genes in breast cancer tissues by vNorthern demonstrated a high expression of oncogenes and low expression of tumor suppressor genes. An open-end analysis of two pools of breast cancer and benign breast tissue libraries by SAGE DGED produced 53 differentially expressed genes according to the screening criteria of a >five-fold difference and p<0.01. Further analysis by EST vNorthern and virtual microarray analysis reduced the candidate genes to six, with four down-regulated genes, ANXA1, CAV1, KRT5 and MMP7, and two up-regulated genes, ERBB2 and G1P3 in breast cancer. These findings were validated by a real-time RT-PCR analysis in eight paired human breast cancer tissue samples. We conclude that the combined multiple high throughput analyses is an effective data mining strategy in cancer gene identification. This approach may improve the usage of public available genomic data through strategic data mining of high throughput analysis.     

p53 and c-erbB-2 protein expression in breast carcinomas. An immunohistochemical study including correlations with receptor status, proliferation markers, and clinical stage in human breast cancer.

Receptor status, proliferative activity, loss of differentiation, inactivation of tumor suppressor genes, and overexpression of oncogenes are related events that may affect the prognosis of patients with breast cancer. Ninety-seven unselected breast carcinomas were immunostained for estrogen and progesterone receptors, Ki-67 proliferation-associated antigen, p53 tumor suppressor gene product (p53), and c-erbB-2 protein. Immunohistochemical results and clinical data were compared. Altered p53 expression (regarded as indirect indication of inactivating gene alterations) was found in 25.8% of cases and was associated with a high Ki-67 labeling index, high mitotic count, and high histologic grade, with c-erbB-2 overexpression, and with negative estrogen and progesterone receptor status. p53 immunostaining could be found also in cytologic samples and correlated with p53 immunoreactivity on frozen sections of the corresponding tumors. c-erbB-2 protein overexpression was seen in 24.7% of cases and was associated with p53 altered expression and negative receptor status. Double immunohistochemical staining showed p53 and c-erbB-2 immunoreactivity in the same cells. Median and mean +/- standard deviation Ki-67 labeling index values were 15 and 16.32 +/- 10.05, respectively. Ki-67 labeling index was correlated with high mitotic count and was positively associated with histologic grade, negative progesterone receptor status, and p53 expression. Estrogen receptor status was not associated with any histologic or clinical parameters, whereas progesterone receptor status was associated with grading. The direct relation of p53 protein alterations with c-erbB-2 overexpression may be interpreted in light of the multistep model of tumor progression. Cases with altered expression of both p53 and c-erbB-2 proteins could be interpreted as having lost one inhibitory control mechanism of cell proliferation and having gained one activator of the malignant potential. However, in comparing cases with the p53 + c-erbB-2 + phenotype with cases showing positivity for only one of these gene products, no association with higher stages was seen. Detection of p53 altered expression on cytologic samples of malignant tumors may have diagnostic relevance, and p53 immunostaining may prove to be an additional diagnostic criterion in cytologic diagnosis.     

Long-term follow-up and prognostic significance of angiogenic basic fibroblast growth factor (bFGF) expression in patients with breast cancer

The development of distant metastasis in breast cancer patients is the key step towards worse prognosis. The angiogenic basic fibroblast growth factor (bFGF) has been associated with tumorigenesis and metastasis in several human cancers. Therefore, bFGF expression was studied by immunohistochemistry in 111 patients with primary breast cancer. The results were correlated with prognostically relevant clinico-pathological features. such as tumor stage, grading. nodal stage and survival. bFGF was expressed in approximately 70% of the breast cancer tissues; 30% of the tumors showed strongly positive staining. With the exception of histological grading (p < 0.05), no correlation was found between the extent of bFGF expression and prognostic parameters. Analysis of survival showed a significantly (p < 0.05) prolonged survival for patients with a concomitant absence of axillary lymph node metastasis and bFGF immunoreactivity. Our data suggest that increased bFGF expression is a novel parameter for worse prognosis in nodal-negative breast cancer patients.     

基于癌基因组图谱挖掘胃癌预后相关基因

目的利用癌基因组图谱数据库中的大量胃癌基因组数据,在高频突变或肿瘤组织高表达的基因中挖掘与临床分化和预后相关的基因。方法从癌基因组图谱数据集中下载胃癌基因组的突变数据,基因表达数据以及病例样本的临床信息资料。筛选并统计出现有害突变频率较高的基因,并比较这些基因在不同分化程度病例中的突变次数。同时利用DESeq2对基因表达数据进行差异表达分析,对于在肿瘤组织中高表达的基因做生存分析,使用Log-rank检验并做FDR校正,筛选出mRNA表达量与胃癌预后相关联的基因。结果突变分析得到PIK3CA和APC的有害突变频率在不同分化程度的胃癌存在差异(Fisher's exact test:P0.05)。差异表达分析得到2 040个在肿瘤组织中上调的基因,2 357个在肿瘤组织中下调的基因。生存分析得到8个对胃癌预后有影响的基因,1个为保护因素,7个危险因素。结论本研究通过挖掘癌基因组图谱中的胃癌数据集,得到了ANGPT2、MMP10和WISP3等与胃癌分化及预后相关的基因,为接下来的研究提供了线索和依据,也为临床治疗提示了新的预后指标。     

cyclinD1,Rb基因蛋白在乳腺癌中的表达

目的：探讨ｃｙｃｌｉｎＤ１基因及Ｒｂ基因的表达与乳腺癌发生发展的关系。方法：应用ＡＢＣ免疫组化法检测２４例良性乳腺组织及５８例乳腺癌中ｃｙｃｌｉｎＤ１及Ｒｂ蛋白表达。结果：乳腺癌中ｃｙｃｌｉｎＤ１过表达阳性率５８．６２％（３４／５８）显著高于良性乳腺组织中的１６．６７％（４／２４），Ｐ＜０．０５。ｃｙｃｌｉｎＤ１过表达出现于导管原位癌并持续于浸润、转移等进展过程中，与年龄、肿瘤大小、组织学类型及淋巴结状态无相关性，但与组织学分级负相关。乳腺癌中Ｒｂ蛋白表达阳性率为３６．２％（２１／５８），显著低于良性乳腺组织的７５％（１８／２４），Ｐ＜０．０５；未见Ｒｂ失表达与临床病理参数间存在相关性，Ｒｂ表达与ｃｙｃｌｉｎＤ１过表达呈正相关。结论：ｃｙｃｌｉｎＤ１过表达及Ｒｂ失表达是乳腺癌发生中的重要事件且前者是一早期分子事件；ｃｙｃｌｉｎＤ１过表达发挥作用可能部分依赖于Ｒｂ蛋白的存在；提示细胞周期调控异常参与乳腺癌的发生。     

妊娠期乳腺癌相关基因的生物信息学分析

目的通过生物信息分析途径对妊娠期乳腺癌患者与正常人群的差异基因进行分析,从分子水平探讨妊娠期乳腺癌的发病机制。方法从公共数据库基因表达数据库(GEO)中下载妊娠期乳腺癌相关数据集,采用Qlucore Omics Explore(QOE)筛选差异表达基因,用DAIVID、STRING等在线分析工具对差异表达基因进行功能富集分析,信号转导通路分析以及预测蛋白质之间的关系。结果共筛选出148个差异表达基因,其中表达上调24个,下调124个,对其进行生物信息学分析发现,TAGLN、ACTG2、TPM2、TPM3、MYLK、ACTA2、MTH11等基因以及MAPK信号通路、黏着斑信号通路、血管平滑肌细胞收缩信号通路等在妊娠期乳腺癌的发生发展中可能起着重要作用。通过STRING分析发现,20个基因处于核心节点位置。结论利用生物信息学的方法能有效分析基因芯片数据并获取生物内在信息。