Tenascin在宫颈鳞癌组织中的表达及其意义

目的　探讨Tenascin(TN)在宫颈鳞癌组织中的表达及其意义。方法　采用免疫组化方法检测了10例正常宫颈、15例宫颈上皮内瘤 (CIN)及 4 8例宫颈鳞癌活检和术后标本的TN表达。结果　TN表达在正常宫颈 +CIN组与宫颈癌组间 (P <0 .0 0 1)、高分化鳞癌与中低分化鳞癌 (P <0 .0 5 )、盆腔淋巴结阳性与阴性组间 (P <0 .0 1)统计学有显著性差异。结论　TN表达可能对宫颈鳞癌间质浸润的评估有指导性意义 ,而且TN染色增强可能预示着生物防御机制的建立和良好的预后。     

Gene expression profiles in human gastric cancer: expression of maspin correlates with lymph node metastasis

To seek for a candidate gene that would regulate tumour progression and metastasis in gastric cancer, we investigated gene expression profiles by using DNA microarray. Tumour tissue and adjacent normal tissue were obtained from 21 patients with gastric cancer and then examined for their gene expression profiles by the Gene Chip Human U95Av2 array, which includes 12 000 human genes and EST sequences. A total of 25 genes were upregulated and two genes were downregulated by at least four-fold in the tumour tissue. In a further analysis according to lymph node metastasis, the expressed levels of maspin, as well as carcinoembryonic antigen and nonspecific crossreacting antigen were significantly higher in tumours with lymph node metastasis than in those without it. Maspin expression in 85 gastric cancer patients was further investigated by using immunohistochemistry. Maspin expression was not observed in normal gastric epithelia without intestinal metaplasia. In contrast, maspin was expressed in 74 of 85 tumour tissues. There was a significant correlation between the incidence of maspin-positive tumour staining and lymph node metastasis. These results suggest that maspin has a potential role for tumour metastasis in gastric cancer.     

Expression of tumour necrosis factor alpha and its receptors in carcinoma of the breast.

The expression of tumour necrosis factor alpha (TNF-alpha) and its two distinct receptors, TNF-R p55 and TNF-R p75, was assessed by immunocytochemistry in 28 primary breast cancer and three reduction mammoplasty specimens (''normal'' breast tissue). Expression of TNF-alpha or TNF-R p75 was not detectable in normal breast tissue or in non-malignant breast tissue adjacent to the tumours. By contrast, TNF-R p55 was expressed by occasional stromal cells in normal tissue. TNF-alpha was expressed focally in 50% of the tumours studied, being largely localised to macrophage-like cells in the stroma. TNF-R p55 was expressed by a population of stromal cells in all the tumours examined, and a varying proportion of neoplastic cells in 75% of these tissues. TNF-R p75 was detected in about 70% of the tumours, immunoreactivity being confined mainly to cells in the stroma. In this preliminary study there was no association between the above cytokine parameters and such measures of tumour biology as lymph node status, tumour grade, proliferative activity or degree of angiogenesis. However, there was a correlation between the expression of TNF-R p55 by blood vessels and the number of leucocytes present.     

Angiogenesis and hypoxia in lymph node metastases is predicted by the angiogenesis and hypoxia in the primary tumour in patients with breast cancer

Hypoxia and angiogenesis are important factors in breast cancer progression. Little is known of hypoxia and angiogenesis in lymph node metastases of breast cancer. The aim of this study was to quantify hypoxia, by hypoxia-induced marker expression levels, and angiogenesis, by endothelial cell proliferation, comparing primary breast tumours and axillary lymph node metastases. Tissue sections of the primary tumour and a lymph node metastasis of 60 patients with breast cancer were immunohistochemically stained for the hypoxia-markers carbonic anhydrase 9 (CA9), hypoxia-inducible factor-1alpha (Hif-1alpha) and DEC-1 and for CD34/Ki-67. Endothelial cell proliferation fraction (ECP%) and tumour cell proliferation fraction (TCP%) were assessed. On haematoxylin-eosin stain, the growth pattern and the presence of a fibrotic focus were assessed. Hypoxia-marker expression, ECP% and TCP% in primary tumours and in lymph node metastases were correlated to each other and to clinico-pathological variables. Median ECP% and TCP% in primary tumours and lymph node metastases were comparable (primary tumours: ECP%=4.02, TCP%=19.54; lymph node metastases: ECP%=5.47, TCP%=21.26). ECP% correlated with TCP% (primary tumours: r=0.63, P<0.001; lymph node metastases: r=0.76, P<0.001). CA9 and Hif-1alpha expression were correlated (primary tumours P=0.005; lymph node metastases P<0.001). In primary tumours, CA9 and Hif-1alpha expression were correlated with DEC-1 expression (P=0.05), presence of a fibrotic focus (P<0.007) and mixed/expansive growth pattern (P<0.001). Primary tumours and lymph node metastases with CA9 or Hif-1alpha expression had a higher ECP% and TCP% (P<0.003); in primary tumours, mixed/expansive growth pattern and fibrotic focus were characterised by higher ECP% (P=0.03). Furthermore, between primary tumours and lymph node metastases a correlation was found for ECP%, TCP%, CA9 and Hif-1alpha expression (ECP% r=0.51, P<0.001; TCP r=0.77, P<0.001; CA9 and Hif-1alpha P<0.001). Our data demonstrate that the growth of breast cancer lymph node metastases is angiogenesis dependent and that angiogenesis and hypoxia in the primary tumour predict angiogenesis and hypoxia in the lymph node metastases. Together with previous findings in breast cancer liver metastases, which grow in 96% of cases angiogenesis independently, these data suggest that both the intrinsic growth characteristics and angiogenic potential of breast cancer cells and the site-specific tumour microenvironment determine angiogenesis and hypoxia in breast cancer.     

Cancer testis antigen expression in primary and recurrent vulvar cancer: association with prognostic factors

Cancer testis tumour associated antigens (C/T-TAAs) were investigated in several gynaecologic and non-gynaecologic neoplasms as possible prognostic markers and targets for immunotherapy. The objective of the present study was to evaluate C/T-TAA expression patterns and prognostic significance in patients affected by vulvar cancer. Melanoma antigen E (MAGE)-A1, MAGE-A4 and NY-ESO-1 expression was determined by immunohistochemistry in paraffin-embedded tissue specimens from 45 primary and 14 recurrent vulvar carcinomas treated with surgery. MAGE-A1, MAGE-A4 and NY-ESO-1 were expressed in 25 (42%), 38 (64%) and 40 (68%) of the 59 samples, respectively. MAGE-A4 was significantly more frequently expressed in tumours with lymph node metastases (p<0.002) and in recurrent tumours (p<0.02). NY-ESO-1 was more highly expressed by moderately or poorly differentiated tumours (p<0.01). This study demonstrates that vulvar cancer frequently expresses C/T-TAAs. Antigen expression correlates with the presence of lymph node metastases and poor tumour differentiation.     

Loss of p21WAF1/CIP1 expression correlates with disease progression in gastric carcinoma.

Previous studies have shown that tumour-suppressor genes play an important role in the progression of solid tumours. Recently, the p21WAF1/CIP1 tumour-suppressor protein has been reported to work as a critical downstream effector of p53 and a potent inhibitor of cyclin-dependent kinases. Thus, the p21WAF1/CIP1 gene is thought to play a central role in tumour suppression. In this study we investigated p21 protein expression in gastric carcinomas. A total of 172 primary gastric carcinoma specimens were immunohistochemically stained for p21 protein expression. Correlations between p21 expression and clinicopathological features were examined. Loss of p21 expression was observed in 104 of 172 tumour tissues (60.4%), and the frequency of p21 loss increased as the stage progressed. Expression of p21 in the primary tumour was frequently lost in patients with either lymph node, liver or peritoneal metastases as compared with patients without metastases. In patients with p21-negative tumours, the risk of recurrence following curative surgery was significantly higher, and the prognosis was significantly poorer than in patients with p21-positive tumours. Loss of p21 expression in primary gastric carcinoma correlates with disease progression. The status of p21 gene expression may have prognostic value in this disease.     

EphB2受体在胃癌中的表达变化及其与转移的关系

目的：观察EphB2在胃癌和转移灶中的表达，并探讨它们在肿瘤发生、发展和转移中的作用以及与胃癌的临床病理参数的关系。方法：采用组织芯片和免疫组化方法检测EphB2在正常粘膜（n=71）、原发癌（n=71）、淋巴结转移癌（n=71）和远处转移癌（n=26）中的表达，比较它们在原发灶和正常组织、转移灶的表达差异。结果：EphB2的表达随肿瘤的发生和进展逐渐下降。EphB2在正常粘膜、原发癌、淋巴结转移和器官远处转移中的表达率分别为：83．1％（59／71）、60．6％（43／71）、43．7％（31／71）和26．9％（7／26）。正常粘膜组织中EphB2的表达率显著高于肿瘤细胞中的表达（P〈0．01）。脏器转移中的表达又显著低于原发癌中的表达（P=0．003）。原发灶和转移灶中EphB2表达不一致的病例数与性别（男性）和浸润深度高度相关（P〈0．05）。结论：肿瘤转移是一个遗传改变逐步积累的结果，EphB2表达下调在胃癌发展及转移中可能起重要作用。     

Upregulation and differential expression of matrilysin (MMP-7) and metalloelastase (MMP-12) and their inhibitors TIMP-1 and TIMP-3 in Barrett's oesophageal adenocarcinoma

Oesophageal adenocarcinoma is believed to arise from metaplastic mucosa in the distal oesophagus, a condition also known as Barrett's oesophagus (BE). BE develops as a result of injury caused by refluxing gastric and duodenal contents and is associated with increased risk of malignant transformation. Matrix metalloproteinases (MMPs) have been implicated in all aspects of tumour progression; tumour growth, basement membrane degradation, invasion and metastatic spread. Using in situ hybridization, we investigated the expression patterns of collagenases-1 and -3, stromelysin-2, matrilysin, metalloelastase and TIMPs-1 and -3 in BE, adenocarcinoma and lymph-node metastases. Matrilysin was expressed abundantly in 12/15 tumours and in 4/6 lymph-node metastases and its expression correlated with the histological aggressiveness of tumour. Matrilysin and metalloelastase were upregulated already in BE. Stromelysin-2 and collagenase-3 expression was detected only in a few tumours. Collagenase-1 was expressed by cancer and stromal cells in 9/15 tumours. Tumour-infiltrating macrophages expressed metalloelastase in 13/15 cancers. TIMPs-1 and -3 were expressed in 12/15 and 11/15 tumours, respectively. Laminin-5 and tenascin were abundantly expressed at the invasive front of poorly differentiated tumours, but not in BE. Our results indicate that matrilysin is the principal MMP expressed by tumour cells in oesophageal adenocarcinoma, and further studies are needed to investigate whether matrilysin or tenascin-C could be used as a predictive marker for progression of BE to cancer.     

SPARC蛋白在胃癌中的表达及与胃癌预后的关系

目的 探讨富含半胱氨酸(Cys)的酸性分泌蛋白(SPARC蛋白)在胃腺癌组织中的表达及与胃腺癌患者预后的关系。方法 免疫组化法检测104例胃腺癌组织和30例癌旁组织中SPARC蛋白的表达差异,并应用统计学的方法分析SPARC蛋白与胃腺癌患者预后的关系。结果 SPARC蛋白在30例癌旁胃黏膜组织中阳性率为20%(6/30),且均为弱表达或无表达,在104例胃腺癌组织中阳性率为76.1%(79/104),二者差异有统计学意义(P＜0.01)。胃腺癌组织中,SPARC蛋白表达高低与淋巴结转移及组织分化程度呈正相关(P＜0.05);SPARC蛋白高表达的胃腺癌患者平均生存时间为27.4个月,明显低于低表达者的40.9个月(P＜0.05)。结论 SPARC蛋白在胃腺癌组织中高表达,且主要表达于胃腺癌细胞周围的间质中;其与淋巴结转移及组织分化程度显著相关;SPARC蛋白高表达的胃腺癌患者生存期短,预后差,但SPARC蛋白不是其独立的预后因素。     

Differential TAG-72 epitope expression in breast cancer and lymph node metastases: a marker of a more aggressive phenotype.

We analysed the differential expression pattern of the three distinct TAG-72 carbohydrate epitopes detected by monoclonal antibodies (MAbs) B72.3, CC83 and CC49 in a consecutive series of 114 patients with primary breast cancer and in 39 synchronous lymph node metastases. B72.3, CC83 and CC49 were expressed in respectively 81 (71%), 68 (60%) and 96 (84%) of the 114 cases. Interestingly, MAb B72.3 was significantly expressed in a subgroup of patients characterised by larger tumour size (p=0.013), lymph node metastasis (p=0.0002), high histopathological grade (p=0.006), high cell kinetics (p=0.04) and advanced clinical stage (p=0.0019). In 20 (51%) of the 39 pairs of matched primary breast cancers and synchronous lymph node metastases, TAG-72 was expressed in the tumour but not in the corresponding metastatic lymph node; tumours with TAG-72-negative lymph nodes appeared to be clinicopathologically more aggressive. CC49, the most immunoreactive and widely used MAb, was not detected in 34% of the metastases of expressing primary tumours. All three MAbs were found in a significantly lower per cent of synchronous metastases with respect to primary tumours (p<0.001).     

Predictive value of cathepsin D and Ki-67 expression at the deepest penetration site for lymph node metastases in gastric cancer

To search for reliable predictors for lymph node metastasis, we immunohistochemically analyzed surgically resected gastric cancer specimens that showed invasion of submucosa (sm) and muscularis propria (mp) of the tumor. The analysis investigated cathepsin D and Ki-67 expression in 136 specimens that were divided into an sm1/sm2 group and an sm3/mp group. In sm1/sm2 group, the incidence of lymph node metastases was significantly higher in tumors with high Ki-67 labeling index (LI) (44%) than in those with low Ki-67 LI (0%). In sm3/mp group, the incidence of lymph node metastases was significantly higher in cathepsin D-positive (56%) and high Ki-67 LI tumors (64%) than in cathepsin D-negative (33%) and low Ki-67 LI (33%). Combined analysis of cathesin D expression and Ki-67 LI correlated strongly with lymph node metastases. No lesions with cathepsin D-negative expression and low Ki-67 LI had lymph node metastases in either group. Cathepsin D and Ki-67 expression may be useful predictors for lymph node metastases in gastric cancer with sm and mp invasion. As predictors, they can identify lesions without lymph node metastases and indicate lesions not needing additional treatment after endoscopic mucosal resection and laparoscopic gastrectomy.     

Contact stimulation of fibroblasts for tenascin production by melanoma cells

Tenascin, an extracellular matrix glycoprotein, is widely expressed in the stroma of almost all types of solid tumours including malignant melanomas. On the basis of its antiadhesive character, it has been supposed that tenascin accumulation facilitates tumour cell invasion and consequent metastasis formation. We aimed to investigate the mechanism by which melanoma cells can modulate the production of tenascin by host stromal cells. The expression of tenascin in cocultures of fibroblasts and five melanoma cell lines, as well as in fibroblast monocultures treated with melanoma conditioned media, was analysed by immunofluorescent staining and image analysis. Tenascin production could not be observed in control fibroblasts or in melanoma cell monocultures. Faint labelling for tenascin could be detected in fibroblast monocultures treated with melanoma cell conditioned media while a very intense staining for tenascin could be seen in melanoma cell-fibroblast cocultures. The tenascin staining in the cocultures was associated with the fibroblasts that were in close contact with melanoma cells. The level of tenascin production around the fibroblasts in different areas of the cocultures correlated well with the density of melanoma cells. Our results indicate that tenascin production of fibroblasts in the tumour stroma is directly modulated by melanoma cells mainly through cell-to-cell contact signalling.     

c-erbB-2 is not a major factor in the development of colorectal cancer

We have investigated c-erbB-2 protein expression in a large cohort of well-characterized colorectal tumours, and in a subset of lymph node metastases. We have also evaluated a Val(655)Ile single nucleotide polymorphism, which is associated with an increased risk of breast cancer, in a subset of the colorectal cancer patients and in healthy control subjects. Immunohistochemical studies revealed that while 81.8% of tumours expressed c-erbB-2, in the majority of cases equivalent levels of c-erb-B2 were seen in adjacent normal mucosa. Colon tumours were significantly more likely to express c-erbB-2 than rectal tumours (P=0.015). Only 52.4% of the metastases displayed staining patterns concordant with their primary tumour, indicating that determination of c-erbB-2 protein in colorectal tumours cannot predict the status of lymph node metastases. PCR--RFLP analysis of the Val(655)Ile single nucleotide polymorphism demonstrated that allele frequencies were identical between colorectal cancer patients and a control group of Caucasian subjects (Ile=0.80 and Val=0.20 in each case), indicating that it is not related to the risk of developing colorectal cancer in this population. Furthermore, there was no relationship between c-erbB-2 protein expression and gene polymorphism (P=0.58). In terms of prognosis, no association was seen between either c-erbB-2 protein expression or the presence of the Val allele and patient survival (P>0.05 in each case), suggesting that c-erbB-2 is not a prognostic marker in colorectal cancer.     

胃癌原发灶和转移淋巴结中酪氨酸磷酸酶受体基因甲基化的差异

目的 研究酪氨酸磷酸酶受体(PTPRG)基因在胃癌原发灶和转移淋巴结中的甲基化差异,以及甲基化抑制剂5-氮脱氧胞苷对于胃癌细胞系甲基化水平的调控功能,进一步阐述胃癌转移的表遗传学机制.方法 应用甲基化特异性PCR(MSP)和逆转录聚合酶链反应(RT-PCR)方法,检测36例胃癌原发灶和转移淋巴结之间PTPRG基因的甲基化差异;检测胃癌细胞系在甲基化抑制剂干预前后PTPRG基因的甲基化及表达调控情况.结果 胃癌原发灶和转移淋巴结PTPRG基因甲基化率分别为25.0%和52.8%,两者PTPRG mRNA表达率分别为50.0%和25.0%,差异均有统计学意义(P＜0.05).胃癌细胞系经甲基化抑制剂干预后,PTPRG基因发生脱甲基化,PTPRG mRNA恢复表达.结论 PTPRG基因在胃癌原发灶和转移淋巴结之间存在明显的甲基化差异.甲基化抑制剂5-氮脱氧胞苷可以降低胃癌细胞系的甲基化水平,恢复PTPRG基因的表达.     

Indium-111-octreotide scintigraphy,intraoperative gamma-detector localisation and somatostatin receptor expression in primary human breast cancer

12 women with primary breast cancer underwent somatostatin receptor scintigraphy (SRS) with ¹¹¹In-DTPA-D-Phe¹-octreotide. The tumour sizes varied between 2 and 5 cm and were all, except one, palpable at clinical examination. Tumour biopsies were taken with additional sampling from normal breast tissue, fat, muscle, axillary lymph nodes and peripheral blood. Ratios between the ¹¹¹In activity concentration in the tissue biopsies (Ti) and in peripheral blood (B) as well as in normal breast tissue (Br) were calculated. In 8/12 patients the scintillation detector was used intraoperatively for radioactivity measurements of the biopsies in situ and ex vivo. The sstr-subtype profiles were determined by northern blot analysis and the relative expression of sstr2 by ribonuclease protection assay (RPA) and immunocytochemistry. Preoperative SRS visualised all primary breast cancer tumours. The scintigraphic image showed no correlation with the histopathological type of the tumour or with the abundance of oestrogen/progesterone receptors on the tumour. Two patients with a massive tumour infiltration of the lymph nodes had a distinct positive SRS of the ipsilateral axilla. In one patient with three nodal metastases the scintigraphic image of the axilla was weak but visible. Four other patients with a negative axillary scintigraphy had 1–2 lymph node metastases. The Ti/B ratios for the breast tumours varied between four and 33 and were not different from Ti/Br ratios. In lymph node metastases the Ti/B ratios were higher (10–41). Intraoperative detector measurements showed a significant difference between the breast tumour and normal tissue in 2/8 patients in situ. Similar measurements on excised tissues (ex vivo) showed a significant difference in 6/8 patients. Two patients with lymph node metastases exhibited a significantly increased uptake ex vivo by detector measurements, but in only one of them in situ. All tumour biopsies expressed the presence of sstr1, 3, 4 and 5, but not of sstr2 at northern analysis. On the other hand, sstr2 was detected in all tumours by RPA and immunocytochemistry. Preoperative SRS visualised primary breast cancer lesions in all 12 patients. SRS could also demonstrate extensive axillary tumour infiltration. Intraoperative use of the scintillation detector could not exclude axillary metastases in situ. The low Ti/B values of both primary tumours and metastases indicate limitations of the radiopharmaceutical used.     

Human epidermal growth factor receptor-2 expression in primary and metastatic gastric cancer

Background

Amplification and overexpression of human epidermal growth factor receptor-2 (HER-2) has been shown in subgroups of gastric cancer, correlated to more aggressive disease and predictive for the treatment with HER-2 antibodies. In this study, we examined the prognostic value of HER-2 expression in primary gastric cancer and in associated lymph node metastases and confirmed the role of HER-2 in tumor angiogenesis by examining vascular endothelial growth factor (VEGF) expression.

Methods

Immunohistochemistry was used to detect HER-2 and VEGF expression in 110 gastric cancer specimens and associated lymph node metastases and in 96 specimens of normal gastric mucosa.

Results

The expression level of HER-2 in gastric tissues was significantly higher than in normal tissues (19.1 % vs. 8.3 %; P < 0.05). HER-2 overexpression was homogeneous in primary gastric cancer and metastatic lymph nodes (P = 0.607). There was a significant positive correlation of HER-2 expression and VEGF expression (P = 0.007). HER-2 overexpression in primary tumor correlated with lymph node metastasis, distant metastasis, and American Joint Committee on Cancer (AJCC) stage. Cox regression multivariate analyses confirmed that tumor size, histological grade, lymph node ratio, AJCC stage, chemotherapy, and HER-2 expression were all prognostic factors. Patients with HER-2 positivity in both primary and metastatic tissues (+/+) had the poorest survival (OS, 12.5 months; DFS, 11.0 months) (P < 0.01).

Conclusions

HER-2 was significantly overexpressed in gastric cancer versus normal tissue and correlated with VEGF expression. HER-2 in tumor or lymph nodes was an independent negative prognostic factor.     

Galectin-1蛋白及孕激素受体与胃癌腹膜转移的关系

目的：探讨胃癌组织中半乳糖结合蛋白（Galectin-1）及孕激素受体（PR）表达与胃癌腹膜转移的关系。方法：应用即用型孕激素测定试剂盒及鼠抗人Galectin-1单克隆抗体和免疫组织化学SP染色法检测40例同期伴腹膜转移胃癌患者的癌旁正常胃黏膜、胃癌原发灶、胃癌腹膜转移灶及淋巴结转移灶中Galectin-1蛋白及PR的表达。结果：正常胃黏膜中Galectin-1及PR的表达与胃癌原发灶、腹膜转移灶及淋巴结转移灶之间差异均具统计学意义（P〈0．05）。胃癌原发灶、腹膜转移灶及淋巴结转移灶三者间的Galectin-1及PR表达差异无统计学意义（P〉0．05）。结论：胃癌病灶中的Galectin-1及PR表达可能成为判断胃癌患者术前是否有腹膜转移及预后的生物学指标,并可为胃癌的内分泌治疗提供依据。