乳腺癌微环境成纤维细胞对乳腺癌细胞表达TIGAR 和Bcl-2 的影响

目的:探索乳腺癌微环境成纤维细胞对乳腺癌细胞表达TIGAR 和Bcl-2 的影响及对乳腺癌生长的作用。方法:体外实验,建立了人乳腺癌细胞株MDA-MB-231 和人成纤维细胞株CCC-ESF-1 共培养模型,RT-qPCR 和Western blot 检测成纤维细胞对乳腺癌细胞表达TIGAR 和Bcl-2 的影响,Annexin V 流式细胞术和Caspase-3 活性荧光检测乳腺癌细胞的凋亡;体内实验,建立人乳腺癌荷瘤裸鼠模型,测量荷瘤裸鼠肿瘤体积,免疫组化检测移植瘤组织TIGAR 和Bcl-2 的表达。结果:体外实验研究结果显示,共培养的成纤维细胞可上调MDA-MB-231 细胞TIGAR 和Bcl-2 的表达并抑制MDA-MB-231 细胞的凋亡;体内实验研究结果显示,与乳腺癌细胞共植入的成纤维细胞能上调荷瘤裸鼠乳腺癌组织TIGAR 和Bcl-2 的表达,高表达的TIGAR 和Bcl-2 可加速荷瘤裸鼠乳腺癌组织生长。结论:乳腺癌微环境成纤维细胞可上调乳腺癌细胞TIGAR 和Bcl-2 的表达,高表达的TIGAR 和Bcl-2 抑制乳腺癌细胞的凋亡,促进了乳腺癌的生长。     

Percoll离心结合免疫磁珠分选的方法从外周血分离单核细胞

目的 探讨采用Percoll密度梯度离心结合免疫磁珠分选从人外周血富集白细胞层中分离提纯单核细胞的方法.方法 采用Ficoll密度梯度离心分离淋巴细胞得到外周血单个核细胞,Percoll密度梯度离心从外周血单个核细胞中富集单核细胞,免疫磁珠阴性分选纯化单核细胞.流式细胞术检测细胞CD14和CD16的表达,分析单核细胞分...     

维生素D通过下调谷氨酰胺合成酶抑制乳腺癌细胞增殖

目的 探究维生素D对非三阴性乳腺癌(non-triple negative breast cancer, Non-TNBC)和TNBC癌细胞增殖的影响及分子机制。方法 收集TNBC和Non-TNBC患者乳腺组织,原代培养TNBC和Non-TNBC患者乳腺细胞;免疫荧光检测乳腺细胞雌激素受体(ER)、孕激素受体(PR)和HER2蛋白表达;免疫荧光检测Non-TNBC和TNBC乳腺癌组织和乳腺细胞中VDR蛋白表达;CCK-8检测细胞活力变化;流式细胞术检测细胞增殖和细胞周期的变化;光学比色法检测细胞谷氨酰胺合成酶(glutamine synthetase, GS)活力的变化;ELISA检测细胞培养上清和胞内谷氨酰胺的水平;Western blot检测细胞GS和VDR蛋白表达的变化;CHIP-PCR检测VDR对GS的转录调控。结果 TNBC患者外周血中维生素D水平和癌组织VDR蛋白表达明显低于Non-TNBC患者(P<0.05)。相比于原代Non-TNBC乳腺癌细胞,原代TNBC患者乳腺癌细胞低表达雌激素受体(ER)、孕激素受体(PR)和HER2蛋白表达;TNBC乳腺癌细胞VDR表达水...     

白介素-13 对与乳腺癌细胞共培养的成纤维细胞SDF-1 和EGF 表达影响

目的:为了探索白细胞介素-13(Interleukin-13,IL-13)在乳腺癌发展中的作用机制,我们研究了IL-13 对与乳腺癌细胞共生长的成纤维细胞表达SDF-1(Stromal cell derived factor 1)和EGF(Epidermal growth factor)的影响。方法:采用体外和荷瘤裸鼠体内人乳腺癌细胞株MDA-MB-231 和人成纤维细胞株CCC-ESF-1(ESF)共培养的方法,用定量PCR(RT-qPCR)方法、流式细胞术和Western blot 方法检测在IL-13 作用下体外共培养的成纤维细胞SDF-1 和EGF 的表达,细胞增殖实验Cell counting kit-8(CCK-8)观察IL-13 对体外共培养的乳腺癌细胞增殖的影响;免疫荧光激光共聚焦显微镜观察在IL-13 作用下荷瘤裸鼠体内与乳腺癌细胞共生长的成纤维细胞SDF-1 和EGF 的表达,检测IL-13 对荷瘤裸鼠肿瘤体积的影响。结果:IL-13 上调体外与乳腺癌细胞共培养的成纤维细胞SDF-1 和EGF 的表达,并促进共培养的乳腺癌细胞的增殖;IL-13 上调荷瘤裸鼠乳腺癌组织成纤维细胞SDF-1 和EGF 的表达,并促进荷瘤裸鼠肿瘤生长。结论:IL鄄13 上调与乳腺癌细胞共培养的成纤维细胞SDF-1 和EGF 的表达,IL-13 对乳腺癌促进作用的分子机制涉及乳腺癌基质成纤维细胞的SDF-1 和EGF。     

丙戊酸钠在γδT细胞对肺癌A549细胞免疫中的作用

目的:研究丙戊酸钠(Sodium Valproate,VPA)作为组蛋白脱乙酰酶抑制剂对肺癌A549细胞MICA蛋白的影响,并探讨丙戊酸钠在人外周血γδT细胞对肺癌A549细胞免疫中的作用.方法:用Ficoll密度梯度离心法分离健康人外周血单核细胞(PBMC),用流式细胞仪分选纯化γδT细胞,用IL-2及唑来磷酸刺激γδT细胞扩增.用不同浓度的丙戊酸钠处理肺癌A549细胞,培养24小时后用PCR及Western blot检测MICA的变化.将分离纯化的γδT细胞与处于对数期生长的肺癌A549细胞混合培养,并加入VPA、MICA抗体处理,24小时后用LDH法检测γδT细胞的细胞毒性.结果:加入VPA后,PCR及Western blot结果显示肺癌A549细胞MICA蛋白在转录及表达上均增强；分离纯化的γδT细胞经体外扩增后,对肺癌A549细胞有较强的细胞毒性作用；加入VPA后γδT细胞对肺癌A549细胞的细胞毒性作用较未加入组增强(P＜0.05).结论:体外扩增的γδT细胞对肺癌A549细胞产生较强的细胞毒性杀伤作用；组蛋白脱乙酰酶抑制剂-丙戊酸钠可上调A549细胞MICA的表达,并增强γδT细胞对肺癌A549细胞的杀伤作用.     

ImmunoAIF△1-480融合蛋白对HER2阳性肿瘤细胞的杀伤作用

目的:观察免疫促凋亡分子ImmunoAIF△1-480对HER2阳性肿瘤细胞的杀伤作用.方法:利用PCR的方法将九聚精氨酸编码序列(R9)与AIF C-末端结构域编码区重组, 然后将R9-AIF△1-480基因与pCMV-e23sFv载体重组, 构建免疫促凋亡分子ImmunoAIF△1-480真核表达载体.用脂质体法转染CHO细胞, 经G418筛选, 建立稳定转染的细胞株, 通过RT-PCR、Western blot、流式细胞术(FCM)等方法检测重组基因在转染细胞中的表达及其对HER2阳性肿瘤细胞的特异性杀伤活性.结果:经过酶切鉴定与测序证实, 带有ImmunoAIF△1-480基因的真核表达载体构建成功, 经RT-PCR、Western blot可检测到培养上清中免疫促凋亡基因的表达, 用FCM检测发现融合蛋白ImmunoAIF△1-480对HER2阳性肿瘤细胞SGC-7901和SKBR-3有明显的促凋亡活性, 而对不表达HER2分子的ECV-304细胞几乎没有影响.结论:ImmunoAIF△1-480可以特异性杀伤HER2阳性肿瘤细胞.     

人卵巢癌荷瘤裸鼠肿瘤微环境局部F4 / 80 阳性巨噬细胞表达Foxp3

目的:探讨人卵巢癌裸鼠腹腔荷瘤模型体内F4/80阳性巨噬细胞是否表达Foxp3,初步探索卵巢癌改造免疫微环境促进自身发展的新机制。方法:应用人卵巢癌细胞系SKOV3构建人卵巢癌裸鼠腹腔荷瘤模型为实验组,以未处理裸鼠为对照组,流式细胞术检测两组外周血和腹水中F4/80阳性巨噬细胞Foxp3的表达情况。应用流式细胞分选两组腹水中F4/80阳性巨噬细胞,分别应用Western blot和PCR检测该细胞中Foxp3在蛋白和mRNA水平的表达情况。免疫荧光双染法鉴定腹腔内卵巢癌组织局部浸润F4/80阳性巨噬细胞Foxp3的表达情况。结果:流式细胞术检测两组外周血中F4/80阳性细胞均不表达Foxp3,差异无统计学意义(P0.05)。流式细胞术和Western blot显示实验组腹水F4/80阳性巨噬细胞在蛋白水平明显表达Foxp3,实时荧光定量PCR显示实验组在mRNA水平明显表达Foxp3,与对照组比较差异均有统计学意义(P0.05)。免疫荧光双染法显示卵巢癌组织可见F4/80和Foxp3双阳性细胞。结论:卵巢癌肿瘤微环境中F4/80阳性巨噬细胞表达Foxp3,为寻找卵巢癌免疫疗法新靶点奠定基础。     

成人外周血平滑肌祖细胞的体外培养

背景：外周血平滑肌祖细胞具有向平滑肌细胞分化的能力。 目的：探索体外分离培养成人外周血平滑肌祖细胞的方法及其生长分化特性。 方法：采用密度梯度离心法分离获得成人外周血单个核细胞,培养12 d后,应用流式细胞仪鉴定分析平滑肌祖细胞并分选纯化,继续培养诱导分化。采用倒置显微镜观察平滑肌祖细胞的形态变化,免疫荧光染色法观察其α-肌动蛋白的表达,同时应用Western blot法检测调宁蛋白、平滑肌肌球蛋白重链的表达情况。此外,观察平滑肌祖细胞的生长特性,绘制生长曲线。 结果与结论：成人外周血单个核细胞诱导培养4 d时开始出现细胞集落,12 d时细胞呈明显梭形。流式细胞仪分析显示,CD14和CD105双阳性的平滑肌祖细胞占贴壁细胞的(71.8±7.2)%。分选纯化的平滑肌祖细胞培养到28 d呈旋涡状生长。间接免疫荧光染色显示,α-肌动蛋白表达呈阳性。Western blot检测显示,调宁蛋白和平滑肌肌球蛋白重链分别于14, 21 d开始表达,并逐渐增加。生长曲线表明,细胞生长至第6天进入对数生长期,第12天后进入平台期。提示通过对外周血单个核细胞的诱导培养,可以获得大量的平滑肌祖细胞。它能稳定增殖,并可进一步分化为平滑肌细胞。     

TGM2 对甲状腺癌干细胞紫杉醇化疗敏感性的影响及作用机制

目的:探究转谷氨酰胺酶2(TGM2)对甲状腺癌干细胞(TCSC)紫杉醇化疗敏感性的影响及作用机制。方法:收集甲状腺癌患者新鲜肿瘤组织,分离培养原代甲状腺癌细胞,经无血清培养成悬浮肿瘤细胞球时,流式细胞仪筛选出CD133~+表型的TCSC进行后续实验。Western blot检测TCSC中干性基因Nanog、Sox2蛋白表达水平。小干扰RNA技术转染TCSC,分为si-TGM2组和si-NC组,MTS检测TGM2 siRNA转染的TCSC对紫杉醇的敏感性;qRT-PCR法检测紫杉醇处理TCSC后细胞TGM2 mRNA表达。软琼脂克隆实验检测TGM2 siRNA转染对紫杉醇处理的TCSC肿瘤球形成能力的影响;Tunnel凋亡和流式细胞仪检测TGM2 siRNA转染对紫杉醇处理的TCSC凋亡的影响。裸鼠实验检测TGM2对紫杉醇处理TCSC敏感性的影响。Western blot检测TGM2 siRNA转染对紫杉醇处理的TCSC凋亡蛋白caspase-3、Bcl-2表达的影响。结果:流式细胞仪分选出CD133~+细胞占全部细胞的1.21%,CD133~+细胞中Nanog、Sox2蛋白表达水平显著高于CD133~-细胞;沉默TGM2表达可显著提高TCSC对紫杉醇的敏感性;紫杉醇处理的TCSC中TGM2 mRNA表达水平显著高于未经紫杉醇处理的TCSC;沉默TGM2表达显著抑制紫杉醇处理的TCSC肿瘤球形成能力,显著增加紫杉醇处理的TCSC凋亡,提高TCSC对紫杉醇的敏感性,提高紫杉醇处理的TCSC凋亡蛋白caspase-3、Bax表达(P0.05)。结论:TGM2基因可能通过抑制凋亡信号通路,降低TCSC的紫杉醇化疗敏感性。     

癌基因DEK和转录因子AP-2α协同促进乳腺癌中HER2的高表达

目的探讨癌基因DEK与转录因子AP-2α在乳腺癌变中的相互作用及在HER2过量表达、乳腺癌变中的病理学意义。方法用Western blot检测组织中DEK、AP-2α和HER2蛋白水平之间的相关性;免疫共沉淀检测DEK和AP-2α在MDA-MB-453乳腺癌细胞中的相互作用;通过siRNA抑制MDA-MB-453细胞中DEK和AP-2α的表达,用半定量RT-PCR和Western blot检测HER2的表达。结果乳腺癌组织中DEK、AP-2α和HER2的蛋白水平之间显示一定的相关性,DEK和AP-2α在MDA-MB-453细胞内有相互作用,siRNA抑制DEK和AP-2α的表达可协同抑制MDA-MB-453细胞中HER2mRNA和蛋白的表达。结论癌基因DEK和转录因子AP-2α协同促进乳腺癌中HER2的高表达。     

A case of metastatic breast cancer with outgrowth of HER2-negative cells after eradication of HER2-positive cells by humanized anti-HER2 monoclonal antibody (trastuzumab) combined with docetaxel

A 38-year-old woman with cancer of the left breast underwent a modified radical mastectomy with lymph node dissection. Twenty-one months later, massive liver metastases and pleural carcinomatosis occurred. The liver metastases responded completely to chemotherapy with trastuzumab combined with docetaxel, but the pleural carcinomatosis was refractory to the therapy. Fluorescence in situ hybridization showed that both the primary tumor and the metastatic tumors of the lymph nodes were composed of HER2 amplification-positive and HER2 amplification-negative cancer cells. This analysis also detected a single cell with HER2 amplification in the pleural effusion that was taken at the completion of the chemotherapy, but four follow-up tests showed no amplified cells. It is speculated that in the liver metastases, the trastuzumab was cytotoxic to both HER2-amplified and nonamplified cancer cells and may have acted through its antiangiogenic effect. However, in the pleural effusion, the effect of trastuzumab was more specific to HER2-amplified cells and caused outgrowth of cancer cells lacking expression of HER2 receptors.     

胸腹水细胞端粒酶活性定性和定量检测

目的探讨端粒酶活性定量检测在诊断良恶性胸腹水中的应用价值。方法采用TRAP-银染定性方法和rrRAP-PicoGreen定量方法,对102例已确诊患者的胸腹水细胞进行端粒酶活性分析。结果恶性胸腹水细胞端粒酶活性明显高于良性胸腹水细胞,其定性检测诊断率明显高于细胞病理学。乳腺癌患者胸腹水细胞的端粒酶活性明显高于卵巢癌、肝癌患者胸腹水细胞的端粒酶活性;肺癌患者胸腹水细胞端粒酶活性明显高于肝癌。在良性胸腹水中,感染性胸腹水细胞端粒酶活性高于非感染性胸腹水。结论恶性胸腹水细胞端粒酶活性明显升高。端粒酶活性定量检测较定性检测更敏感、简便,对良恶性胸腹水的诊断和鉴别诊断有一定应用价值。     

Clinical evaluation and tissue distribution of CA125 in patients with pleural effusion

CA125 in serum and pleural effusion was measured in 51 patients with malignant effusion and 38 patients with benign effusion, and the tissue distribution of CA125 was investigated by immunohistochemical technique. The 51 malignant effusions were secondary to primary lung cancer. The 38 benign effusions were taken from 23 patients with tuberculous pleurisy, 9 patients with empyema, 5 patients with congestive heart failure and one patient with nephrosis. In the mean level and the positive rate of serum CA125, no significant difference was shown between primary lung cancer and tuberculosis or the other benign diseases. The mean level of CA125 in pleural effusion of primary lung cancer was significantly higher than that in pleural effusion of tuberculosis (p less than 0.01), and showed a tendency to increase compared to that in pleural effusion of the other benign diseases (p less than 0.1). The mean level of CA125 in pleural effusion of tuberculosis was significantly lower than that in the other benign diseases (p less than 0.02). The positive rate of CA125 in malignant effusion was 43.1% and the diagnostic specificity of it was 86.7%. CA125 was detected in carcinoma cells and activated mesothelial cells in pleural effusion and mesothelial cells of normal pleural tissue by immunohistochemical staining. These results suggest that the measurement of CA125 in pleural effusion is useful for differential diagnosis of the malignant effusion from the benign effusion and that CA125 in pleural effusion of pleuritis carcinomatosa is produced by not only carcinoma cells but also activated mesothelial cells.     

Overexpression of HER2/neu in solid tumours: an immunohistochemical survey

AIMS: Using a standardized immunohistochemical assay we have evaluated 575 primary neoplasms of different histogenesis to determine the incidence of HER2 overexpression in some of the most common categories of human solid neoplasms. This study addresses the variable incidence of HER2 overexpression previously published for some tumour types. METHODS AND RESULTS: The immunohistochemical staining was performed on paraffin sections of surgical specimens and a well-defined scoring system based upon numbers of HER2 receptors expressed on the cell surface was applied. Overexpression of HER2 as defined as a HER2 score of equal or greater than 2 was seen in breast cancer (22%), pulmonary adenocarcinoma (28%), colorectal adenocarcinomas (17%), pulmonary squamous (11%) and gastric adenocarcinomas (11%). As expected, the proportion of cases with a HER2 score of 3 was highest in breast cancer. Contrary to published results prostate and pancreas adenocarcinomas showed a very low incidence of HER2 overexpression. CONCLUSIONS: Overexpression of HER2 is detected immunohistochemically in a proportion of epithelial neoplasms of diverse histogenesis in addition to ductal breast cancer. The standardized format of the assay will allow comparative analyses of studies performed at different institutions.     

Expression of wild-type estrogen receptor beta protein in human breast cancer: specific correlation with HER2/neu overexpression

Expression of estrogen receptor beta (ERbeta) protein in human breast cancer and correlation with clinicopathological factors have been reported by many investigators, but many of them used ERbeta antibodies that react with both wild-type ERbeta (ERbetawt) and splicing variant isoform. Therefore, the frequency and correlation with clinicopathological factors of ERbetawt expression remain to be established. In the present study a monoclonal antibody EMR02, specific for ERbetawt, was used in formalin-fixed paraffin-embedded sections from 225 female primary breast cancer patients diagnosed as having invasive ductal carcinoma. Expression of ERalpha, progesterone receptor (PgR) and HER2/neu were also investigated by immunohistochemistry. For ERbetawt, ERalpha and PgR, positivity was defined as nuclear staining in >10% of the cancer cells. HER2/neu overexpression was defined as a Hercep test score 3+. Positivity for ERbetawt, ERalpha, PgR and HER2/neu overexpression was 55%, 74%, 61% and 25%, respectively. The expression of ERbetawt had a positive correlation with ERalpha (P=0.018) and PgR (P=0.02). There was significant positive correlation between ERbetawt expression and HER2/neu overexpression (P<0.0001). According to multivariate logistic regression analysis the most significant association was between ERbetawt expression and HER2/neu overexpression (P<0.0001). These results suggest that clinical significances of ERbetawt expression in human breast cancer patients may be more complex.     

HER2/HER3‐positive metastatic salivary duct carcinoma in the pleural effusion: A case report

Salivary duct carcinoma (SDC) is an aggressive form of salivary gland tumor, and SDC patients tend to be older men, more commonly in advanced stage with a poorer prognosis. Although the cytological characteristics of SDC on fine‐needle aspiration cytology have been well‐described at the primary site, they have not been explored in metastasis. Here we reported a case of HER2/HER3‐positive metastatic SDC in the lung and pleural effusion. The patient was a man in his 50s who had undergone extended total parotidectomy in 2008. He was originally diagnosed as having HER2‐positive left parotid SDC. Six years later a mass was discovered in the left lung by chest computed tomography (CT) and was diagnosed as metastatic SDC by both bronchial biopsy and cytology. Subsequently he had a recurrent SDC in the left pleural effusion and died of respiratory failure. Cytological findings from bronchial brushing smear showed small sheet clusters in a slightly necrotic background. In the pleural effusion cytology, tumor cells appeared as ball‐like clusters of epithelioid cells with apocrine‐like findings. In immunocytochemistry, HER3 of SDC cells in pleural effusion was significantly overexpressed relative to the matched primary tumor, even though HER2 amplification did not change. Cytological findings and HER family receptors differed between the primary and metastatic SDC. Therefore, molecular tests, such as protein expression and gene amplification using cytological specimens, may become important in future when determining therapy strategies in patients with distant metastasis.     

Novel HER2 Aptamer Selectively Delivers Cytotoxic Drug to HER2-positive Breast Cancer Cells in Vitro

ABSTRACT: BACKGROUND: Aptamer-based tumor targeted drug delivery system is a promising approach that may increase the efficacy of chemotherapy and reduce the related toxicity. HER2 protein is an attractive target for tumor-specific drug delivery because of its overexpression in multiple malignancies, including breast, gastric, ovarian, and lung cancers. METHODS: In this paper, we developed a new HER2 aptamer (HB5) by using systematic evolution of ligands by exponential enrichment technology (SELEX) and exploited its role as a targeting ligand for delivering doxorubicin (Dox) to breast cancer cells in vitro. RESULTS: The selected aptamer was an 86-nucleotide DNA molecule that bound to an epitope peptide of HER2 with a Kd of 18.9 nM. The aptamer also bound to the extracellular domain (ECD) of HER2 protein with a Kd of 316 nM, and had minimal cross reactivity to albumin or trypsin. In addition, the aptamer was found to preferentially bind to HER2-positive but not HER2-negative breast cancer cells. An aptamer-doxorubicin complex (Apt-Dox) was formulated by intercalating Dox into the DNA structure of HB5. The Apt-Dox complex could selectively deliver Dox to HER2-positive breast cancer cells while reducing the drug intake by HER2-negative cells in vitro. Moreover, Apt-Dox retained the cytotoxicity of Dox against HER2-positive breast cancer cells, but reduced the cytotoxicity to HER2-negative cells. CONCLUSIONS: The results suggest that the selected HER2 aptamer may have application potentials in targeted therapy against HER2-positive breast cancer cells.     

Utility of CK7 and CK20 immunohistochemistry in the detection of synchronous breast and colon carcinoma in a pleural effusion: a case report and supporting survey of archival material

We present a case of synchronous breast and colon carcinoma in a pleural effusion, to our knowledge the first such reported case in the English-language literature. The patient was a 55-yr-old white female with known metastatic breast and colon carcinoma who developed a malignant pleural effusion which demonstrated two strikingly different populations of malignant cells by immunohistochemical study of cell block material. One cell population demonstrated a cytokeratin (CK)7+/CK20-/ER+ phenotype, while the other demonstrated a CK7-/CK20+/ER- phenotype, consistent with breast and colon origin, respectively. An immunohistochemical survey of archival breast and colon primary and metastatic carcinomas confirmed the established CK7+/CK20- phenotype of breast and CK7-/CK20+ phenotype of colon primary carcinomas, and the maintenance of this phenotype in metastases thereof. A survey of benign and malignant mesothelial lesions confirmed the absence of staining for estrogen receptor, but showed 6/10 cases weakly positive for CK20, which has not been described in other published series. This unusual case graphically illustrates the utility of cytokeratin subset immunohistochemistry in effusion cytology.     

Proteomic analysis of exosomes isolated from human malignant pleural effusions

Exosomes are membrane vesicles from endosomal origin secreted by various cells such as hematopoietic, epithelial, and tumor cells. Exosomes secreted by tumor cells contain specific antigens potentially useful for immunotherapeutic purposes. Our aim was to determine if exosomes are present in human cancerous pleural effusions and to identify their proteomic content. Exosomes were purified by sucrose gradient ultracentrifugation, and electron microscopy was used to check both concentration and purity of exosomes. Proteins were separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and protein bands were identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and Western blotting. Exosomes were present in pleural fluid obtained from patients suffering from mesothelioma (n = 4), lung cancer (n = 2), breast cancer (n = 2), and ovarian cancer (n = 1). As previously reported by others, antigen-presenting molecules, cytoskeletal proteins, and signal transduction-involved proteins were present. Proteins not previously reported were identified (SNX25, BTG1, PEDF, thrombospondin 2). Different types of immunoglobulins and complement factors were abundantly present in the sucrose fractions containing exosomes. Exosome-directed specificity of these immunoglobulins was not observed. In conclusion, sucrose gradient ultracentrifugation allows isolation of exosomes from malignant pleural effusions. However, pleural fluid proteins and especially immunoglobulins are coisolated and may hamper the use of exosomes isolated from malignant effusion for immunotherapy programs.     

Salivary Duct Carcinoma - A Highly Aggressive Salivary Gland Tumor with HER-2/neu Oncoprotein Overexpression

Salivary duct carcinoma (SDC) is a highly malignant salivary gland tumor with aggressive clinical behavior, and is characterized by its histological resemblance to invasive ductal carcinoma of the breast. Overexpression and/or amplification of proto-oncogene Her2/neu has been shown to influence both prognosis and treatment of breast cancer. Since salivary duct carcinoma and ductal breast carcinoma share many common characteristics, HER2/neu overexpression might also be important in SDC. However, data on the expression of c-erbB2/HER2/neu in salivary gland tumors are still scarce. Therefore, we have evaluated 15 cases of salivary duct carcinomas (SDC) for HER2/neu overexpression using immunohistochemistry with the HercepTest. Overexpression, identified as strong or moderate membrane immunostaining, was observed in all but one case of SDC in most neoplastic cells. Thus, our study suggests that anti-HER2/neu therapy with Herceptin is beneficial for patients with aggressive salivary duct carcinoma.