食管癌组织中KAI1、EphA2蛋白的表达及其意义

目的探讨KAI1蛋白和EphA2蛋白的表达与食管癌发生及浸润转移的关系以及两者表达的相关性。方法采用免疫组化方法,检测160例食管癌组织及其150例正常黏膜中KAI1和EphA2蛋白的表达。结果KAI1蛋白在癌组织中的表达明显低于其正常黏膜（P〈0．05）,在有淋巴结转移病例中的表达明显低于无淋巴结转移病例（P〈0．05）,KAI1蛋白的表达与肿瘤的分化程度、浸润深度无关（P〉0．05）。EphA2蛋白在食管癌组织中的表达明显高于其正常黏膜（P〈0．05）,在深层浸润组的表达明显高于浅层浸润组（P〈0．05）,在有淋巴结转移组中的表达明显高于无淋巴结转移组（P〈0．05）,EphA2蛋白的表达与肿瘤的分化程度无关（P〉0．05）。KAI1蛋白和EphA2蛋白的表达在有淋巴结转移的病例中有显著负相关（P〈0．05）。结论 KAI1蛋白的低表达和EphA2蛋白的高表达与食管癌的癌变及转移的发生有关,两者对于食管癌的淋巴结转移具有负向调节作用。     

Clinical significance of p16 protein expression loss and aberrant p53 protein expression in pancreatic cancer

Pancreatic cancer is a disease with poor prognosis mainly due to low resection rates and late diagnosis. To increase resectability and improve survival rates, a better understanding of pancreatic cancer pathogenesis and more effective screening techniques are required. New methods, such as genetic and molecular alterations, may suggest novel approaches for pancreatic cancer diagnosis and treatment. We immunohistochemically investigated 44 formalin-fixed, paraffin-embedded specimens of pancreatic ductal adenocarcinoma using monoclonal anti-p16 antibodies and monoclonal anti-p53 antibodies. The expressions of p16 and p53 proteins were compared using the Chi-square test with SPSS. Disease-free survival was analyzed using the Kaplan-Meier method, verified by the Log- Rank test. Loss of p16 expression was noted in 20 (45.5%) cases and aberrant p53 protein expression was detected in 14 (31.8%) cases. Loss of p16 expression was associated with a higher incidence of lymph node metastasis (p=0.040) and a more advanced stage (p=0.015), although there was no significant correlation between p16 expression and survival. Aberrant p53 protein expression correlated with histologic grade (p= 0.038). Disease-free survival rate was significantly lower in the aberrant p53 protein positive group compared to the negative group (p=0.029). From our results, we suggest that p53 is not a prognostic factor; however, p16 and p53 genes do play important roles in the progression of pancreatic ductal adenocarcinoma.     

De novo expression of EphA2 in osteosarcoma modulates activation of the mitogenic signalling pathway

Fritsche‐Guenther R, Noske A, Ungethüm U, Kuban R‐J, Schlag PM, Tunn P‐U, Karle J, Krenn V, Dietel M & Sers C (2010) Histopathology 57, 836–850 De novo expression of EphA2 in osteosarcoma modulates activation of the mitogenic signalling pathway Aims: In osteosarcoma patients the development of metastases, often to the lungs, is the most frequent cause of death. The aim of this study was to elucidate the molecular mechanisms governing osteosarcoma development and dissemination and, thereby, to identify possible novel drug targets for improved treatment. Methods and results: Osteosarcoma samples were characterized using genome‐wide microarrays: increased expression of the EphA2 receptor and its ligand EFNA1 was detected. In addition, increased expression of EFNB1, EFNB3 and EphA3 was suggested. Immunohistochemistry revealed an absence of EphA2 in normal bone, and de novo expression in osteosarcomas. EFNA1 was expressed in normal bone, but was significantly elevated in tumours. Further in vitro investigations on the functional role of EphA2 and EFNA1 showed that EFNA1 ligand binding induced increased tyrosine phoshorylation, receptor degradation and downstream mitogen‐activated protein kinase (MAPK) activation. Interference with the MAPK pathway unravelled a potential autoregulatory loop governing mainly EFNA1 expression via the same pathway. Conclusion: Upregulation and de novo expression of ephrins in osteosarcomas are involved in oncogenic signalling and thus might stimulate osteosarcoma metastasis.     

EphA2在乳腺癌中的表达及其与上皮间质转化的关系

目的　研究EphA2、上皮间质转化（EMT）在乳腺癌中的表达情况及意义,探讨EphA2与EMT在乳腺癌的侵袭与转移过程中的关系。 方法　采用免疫组织化学方法检测30例乳腺纤维瘤组织和110例乳腺癌中EphA2、E-钙黏蛋白（E-cadherin）、β-连环蛋白（β-catenin）、波形蛋白（vimentin）的表达情况,分析其与乳腺癌临床病理因素的关系及EphA2、E-cadherin蛋白、β-catenin蛋白、vimentin蛋白表达的相关性。 结果　在30例乳腺纤维瘤组织中EphA2、E-cadherin蛋白、β-catenin蛋白、vimentin蛋白阳性率分别为20%、90%、26.67%、13.33%,在110例乳腺癌中的阳性率分别为61.82%、34.54%、64.54% 、68.18%。EphA2的高表达、E-cadherin蛋白的低表达、β-catenin蛋白和vimentin蛋白的高表达与临床分期、淋巴结转移、组织学分级显著相关（P<0.05）,EphA2的高表达与E-cadherin蛋白的低表达呈负相关（P<0.05）,与β-catenin蛋白、vimentin蛋白的高表达呈正相关（P<0.05）。 结论　EphA2、E-cadherin蛋白、β-catenin蛋白、vimentin蛋白与乳腺癌的侵袭与转移有相关性;EphA2可能通过调控E-cadherin蛋白、β-catenin蛋白、vimentin蛋白的表达来促进EMT,进而在乳腺癌的侵袭和转移过程发挥重要作用。     

Expression of EphA2 and EphrinA-1 in vulvar carcinomas and its relation to prognosis

AIMS: To examine the expression of EphA2 and EphrinA-1 in vulvar squamous cell carcinomas and investigate their prognostic relevance. METHODS: Tumours from 224 patients with vulvar squamous cell carcinomas were investigated for expression of EphA2 and EphrinA-1 using single and double immunostaining methods. RESULTS: High expression (strong/moderate staining intensity) of EphA2 and EphrinA-1 was observed in 114 (51%) and 126 (56%) vulvar carcinomas, respectively. In the three cases tested using the double immunostaining method, colocalisation of EphA2 and EphrinA-1 proteins was identified in the same neoplastic cells. High EphA2 expression was significantly correlated to high expression of EphrinA-1 (p<0.01) and cyclin A (p<0.01), large tumour size (p = 0.03), deep invasion (p<0.01) and higher FIGO stage (p = 0.05). A correlation between high EphrinA-1 expression and high levels of cyclin A (p<0.01) and p21 (p<0.01), deep invasion (p<0.01) and higher FIGO stage (p = 0.01) was also seen. In univariate analysis, high expression of EphrinA-1 was associated with poor survival (p = 0.03). However, in the multivariate analysis neither EphrinA-1 nor EphA2 were significantly correlated to survival. CONCLUSIONS: EphA2 and EphrinA-1 were overexpressed in 51% and 56% of the vulvar squamous cell carcinomas, respectively, and high levels of EphA2 and EphrinA-1 proteins were associated with deep tumour invasion and high FIGO stage. However, EphA2 and EphrinA-1 were not independently associated with clinical outcome in vulvar carcinomas.     

Expressions of EphA2 and EphrinA-1 in early squamous cell cervical carcinomas and their relation to prognosis

By using immunohistochemistry we investigated the expression of EphA2 and EphrinA-1 in 217 early squamous cell cervical carcinomas and examine their prognostic relevance. For EphA2 expression, 21 tumors (10%) showed negative, 108 (50%) weak positive, 69 (32%) moderate positive and 19 (9%) strong positive, whereas for EphrinA-1 expression, 33 tumors (15%) showed negative, 91 (42%) weak positive, 67 (31%) moderate positive and 26 (12%) strong positive. In univariate analysis high expression (strong staining) of EphrinA-1 was associated with poor disease-free (P = 0.033) and disease-specific (P = 0.039) survival. However, in the multivariate analyses neither EphrinA-1 nor EphA2 was significantly associated to survival. The increased levels of EphA2 and EphrinA-1 in a relative high number of early stage squamous cell carcinomas suggested that these two proteins may play an important role in the development of a subset of early cervical cancers. However, EphA2 and EphrinA-1 were not independently associated with clinical outcome.     

乳腺癌EphA2和EphrinAl的表达及与临床病理因素的关系

目的　研究云南白族地区乳腺癌EphA2和EphrinAl的表达及其与临床病理因素的关系。 方法　用免疫组织化学(IHC)检测乳腺癌组织中EphA2、EphrinAl的表达,比较各自表达情况与临床病理因素的关系及二者间的相关性。 结果　EphA2、EphrinAl主要表达于肿瘤细胞和血管内皮细胞的胞浆和胞膜,呈棕黄色或棕褐色。150 例乳腺癌组织中,EphA2、EphrinAl阳性表达分别为123例、129例,阳性率分别为82%、86%。二者的阳性率与患者年龄无相关性(P>0.05),而与病理类型、肿瘤大小、淋巴结转移、临床分期和组织学分级有相关性(P<0.05)。浸润性导管癌EphA2和EphrinAl阳性率较导管内癌的高;肿瘤较大组、淋巴结转移组、临床分期较晚者、组织学分级较高组EphA2和EphrinAl的阳性率分别高于肿瘤较小组、无淋巴结转移组、临床分期较早者、组织学分级较低组EphA2和EphrinAl的阳性率。EphA2和EphrinAl阳性染色共同定位于大致相同的肿瘤区域和血管内皮细胞,二者的阳性率有相关性(P<0.05)。 结论　EphA2、EphrinAl在乳腺癌高表达,并与其发生发展、侵袭转移及恶性程度有关,有望成为乳腺癌预后评估标志物,为早期诊断和靶向治疗提供新思路。     

EphA2 as a new target for breast cancer and its potential clinical application

Purpose: The aim of this research was to study the expression of EphA2 to assess its suitability as a new breast cancer target. Methods: Immunohistochemistry (IHC) was used to detect EphA2 protein expression in pathology tissue samples from 250 cases of breast cancer, and the expression of EphA2 mRNA was detected by in situ hybridization (ISH). Breast cancer cells were isolated and cultured. The expression of EphA2 in the cells was detected by the indirect immunofluorescence assay (IFA), and the expression of EphA2 in breast cancer was analysed. Results: EphA2 protein and mRNA were mainly expressed in tumor cells and vascular endothelial cells. EphA2 protein was expressed in 187 cases, with a positive rate of 74.80%, whereas EphA2 mRNA was expressed in 209 cases, with a positive rate of 83.60%. EphA2 protein and mRNA expression were correlated with lymph node metastasis, clinical stage, and breast cancer histologic grade (P<0.05). In addition, the positive expression rates of EphA2 protein and EphA2 mRNA were correlated (P<0.05). EphA2 was barely expressed in normal breast cells but highly expressed in breast cancer cells. Conclusion: EphA2 is highly expressed in breast cancer tissues and has the potential to be a new breast cancer target, providing a preliminary basis for the development of new targeted drugs for breast cancer and the construction of fluorescent-targeted tracers for fluorescence-guided mastoscopic breast-conserving surgery.     

EphA2及其配体在子宫内膜样腺癌中的表达及其与血管生成的关系

 目的： 探讨生促红素肝细胞受体A2 （EphA2）及其配体ephrin-A1在子宫内膜样腺癌组织中的表达及其与肿瘤血管生成的关系。方法： 利用免疫组织化学法检测56例子宫内膜样腺癌、20例子宫内膜增生过长、30例正常子宫内膜增殖期和30例分泌期组织中EphA2、ephrin-A1、雌激素受体（ER）和孕激素受体（PR）的表达,并采用CD34抗体标记微血管内皮细胞,计算微血管密度(MVD)。分析EphA2和ephrin-A1的表达与MVD之间的相关性及其与子宫内膜样腺癌临床病理特征的关系。结果： 子宫内膜样腺癌组织中EphA2和ephrin-A1的表达显著高于子宫内膜样增生过长及正常子宫内膜(P<0.05);EphA2和ephrin-A1表达水平及MVD值与子宫内膜样腺癌FIGO分期、肿瘤分化程度、肌层浸润深度、淋巴微血管浸润和孕激素受体表达有关(P<0.05);Spearman等级相关分析表明EphA2和ephrin-A1表达分别与MVD呈显著正相关(r=0.476,P<0.05;r=0.501,P<0.05)。结论： EphA2及其配体ephrin-A1在子宫内膜样腺癌中高表达,可能参与了肿瘤血管生成和孕激素抵抗。     

KAI1及其等位基因杂合性缺失与胰腺癌转移及预后的相关性

目的 探讨肿瘤转移抑制基因KAI1与胰腺癌转移及预后的相关性,并进一步研究此相关性与其等位基因杂合性缺失(LOH)的关系.方法 收集胰腺癌标本62例,行免疫组织化学Power-vision二步法检测KAI1基因(CD82)的表达;石蜡切片显微切割癌及癌旁正常胰腺组织,提取DNA,变性高效液相色谱法(DHPLC)检测LOH.结果 CD82在胰腺癌原发灶中表达率为76%(47/62),在有淋巴结转移、远处脏器转移病例中表达显著降低(P<0.05),在晚期(Ⅲ、Ⅳ)肿瘤中比早期(Ⅰ、Ⅱ)表达显著降低(P<0.01),CD82表达阳性者的1年生存率显著高于表达阴性者(P<0.05).显微切割标本PCR扩增,微卫星D11S1344有5对、D11S1326有2对存在LOH,发生率为17%.结论 随着胰腺癌的进展,CD82的表达逐渐降低,这种降低与KAI1等位基因的杂合性缺失有关.检测CD82的表达及LOH对判断肿瘤的分期,淋巴转移和远处转移及患者的预后有一定的参考价值.     

云南大理白族地区乳腺癌EphA2和EphrinA1的表达及其意义

目的:研究云南白族地区乳腺癌EphA2、Ephrin A1的蛋白和mRNA的表达及其意义。方法:选取大理大学附属医院2012～2015年乳腺癌组织石蜡标本150例,用免疫组织化学和原位杂交方法分别针对乳腺癌组织中EphA2、Ephrin A1的蛋白及mRNA表达进行检测,分析各自蛋白和mRNA的表达与临床病理因素的关系及它们之间的相关性。结果:EphA2、Ephrin A1的蛋白和mRNA主要表达于肿瘤细胞和血管内皮细胞的胞质和胞膜。EphA2、Ephrin A1蛋白阳性率分别为82%和86%;mRNA阳性率分别为60.67%和65.33%。EphA2、Ephrin A1的蛋白和mRNA在乳腺癌的阳性率与病理类型、肿瘤大小、淋巴结转移、临床分期和组织学分级相关。浸润性导管癌EphA2和Ephrin A1的蛋白及mRNA的阳性率较导管内癌高;肿瘤较大组、淋巴结转移组、临床分期较晚者、组织学分级较高组EphA2、Ephrin A1的蛋白及mRNA的阳性率分别高于肿瘤较小组、无淋巴结转移组、临床分期较早者、组织学分级较低组的。EphA2、Ephrin A1蛋白阳性染色共同定位于大致相同的肿瘤区域和血管内皮细胞,两者的阳性率有相关性;EphA2、Ephrin A1 mRNA的表达具有与蛋白表达类似的特征,两者的阳性率也有相关性。乳腺癌中EphA2、Ephrin A1各自蛋白与其相应mRNA的阳性表达相关,但表达水平不完全一致。结论:乳腺癌中EphA2、Ephrin A1的蛋白及mRNA高表达,并与其发生发展、侵袭转移及恶性程度有关,有望作为乳腺癌早期诊断及预后判断的标志物,作为基因治疗新靶向。     

Correlation and prognostic significance of MMP-2 and TFPI-2 differential expression in pancreatic carcinoma

Aberrant expression of matrix metalloproteinase (MMP)-2 and tissue factor pathway inhibitor (TFPI)-2 not only correlate with tumorigenesis, but also with tumor invasion and metastasis. This study aims to investigate the correlation and prognostic significance of MMP-2 and TFPI-2 differential expression in pancreatic carcinoma. Immunohistochemistry was used to evaluate MMP-2 and TFPI-2 expression in tumor tissues and corresponding non-tumor tissues from 122 patients with pancreatic carcinoma. The results showed that the expression of MMP-2 was significantly (P < 0.05) higher in tumor tissues (78.7%) than in adjacent non-tumor tissues (27.9%), whereas the expression of TFPI-2 was significantly (P < 0.001) lower in tumor tissues (27.9%) than in adjacent non-tumor tissues (79.5%). Spearman’s rank correlation test showed a negative correlation between MMP-2 and TFPI-2 expression (r = -0.346, P < 0.001). Kaplan-Meier survival analysis showed that high MMP-2 expression was significantly correlated with decreased disease-free survival (DFS) (P < 0.001) and overall survival (OS) (P < 0.001), while high TFPI-2 expression was significantly associated with increased DFS (P < 0.001) and OS (P < 0.001) of the patients. Multivariate analysis showed that high MMP-2 expression can act as an independent predictive factor for poor DFS (P = 0.01); and low TFPI-2 expression as an independent prognostic factor for poor DFS (P < 0.001) and OS (P < 0.001). In conclusion, our findings suggested that the differential expression of MMP-2 and TFPI-2 have a negative correlation in pancreatic carcinoma tissues; they may be considered as valuable biomarkers for prognosis of pancreatic carcinoma.     

A comprehensive in vitro characterization of pancreatic ductal carcinoma cell line biological behavior and its correlation with the structural and genetic profile

There are a large number of stable pancreatic ductal carcinoma cell lines (PDCL) that are used by researchers worldwide. Detailed data about their differentiation status and genetic alterations are present in the literature, but a systematic correlation with cell biological behavior is often lacking. PDCL (n=12) were clustered by source of tumor cell (ascites, primary tumor, metastasis), and the data of functional cell biology were correlated with the reported structural and genetic profiles. Major histocompatibility complex expression, chemosensitivity and aneuploidia appeared to be related to the source of PDCL, and proliferative capacity appeared to be related to the grade of differentiation. No correlation between genetic/structural features of PDCL and biological behavior was found. All the cell lines appeared generally insensitive to in vitro treatment with 5-fluorouracil and showed variable degrees of susceptibility to gemcitabine, raltitrexed and oxaliplatin. All the PDCL showed resistance to Fas-mediated apoptosis but were significantly sensitive to the pro-apoptotic effect of inflammatory cytokines [interleukin (IL)-1, tumor necrosis factor (TNF) and interferon ]. PDCL were characterized for the secretion of several factors relevant to the tumor-immune cross talk. Vascular endothelial growth factor, CCL2, CCL5 and transforming growth factor were the factors most frequently released; less frequent was the secretion of CXCL8, CCL22, IL-6 and sporadically CXCL12, IL-10 and hepatocyte growth factor. The cytokines IL-1 and TNF were always undetectable. In conclusion, a clear correlation between structural/genetic features and function could not be detected, suggesting the weakness of a morphological classification for the in vitro studies of pancreatic cancer.     

P53/MDM2 overexpression in metastatic endometrial cancer: correlation with clinicopathological features and patient outcome

Several studies have reported that p53/mdm2 distortions play a pivotal role in the development and progression of various human malignancies. However, the number of reports having evaluated simultaneously the components of the P53-pathway alterations in advanced-stage human endometrial carcinomas (EC) is low. In this study, we examined the expression of P53/MDM2 proteins in primary and metastatic ECs, and analyzed the clinicopathological characteristics as well as the survival outcome of patients in relation to P53/MDM2 overexpression. The study group comprised 36 patients with advanced EC, whose primary and metastatic tumor slides were sufficient for analysis. Immunohistochemical assessment was made by applying anti-human P53 and MDM2 antibodies and a highly sensitive EnVision⁺/HPR visualization system. Nuclear P53 overexpression was seen in 11 (31%) primary ECs and 12 (33%) metastatic tumors. There was a significant correlation between P53 overexpression (in primary cancers and metastatic tumors) and MDM2 overexpression in metastatic tumors. Nuclear MDM2 overexpression was noted in 42% (15/36) of primary carcinomas and in 47% (17/36) of metastatic tumors. A significant association existed between MDM2 overexpression and histological grading (G1 + G2 versus G3, P = 0.043). P53/MDM2 overexpression occurred simultaneously in 7 out of 36 (19%) primary ECs and in 9 out of 36 (25%) metastatic lesions. Concomitant overexpression of these proteins was reported in 7 out of 36 (19%) cases and tended to be higher in tumors showing VSI compared to neoplasms lacking vascular space invasion (P = 0.051). P53 overexpression, either in primary ECs (P < 0.0001) or metastatic lesions (P < 0.0001), was significantly associated with poor survival in univariate analysis. Moreover, the log-rank test demonstrated that simultaneous P53/MDM2 overexpression was also correlated with decreased length of survival. There was no correlation between MDM2 overexpression and patient survival. Multivariate Cox regression analysis revealed that only P53 overexpression is an independent predictor of survival. In conclusion, our data support the view that patients with P53 overexpression are significantly associated with an unfavorable outcome, whereas MDM2 overexpression is not related to decreased survival length in women operated on for advanced-stage EC.     

Clinicopathologic correlation of beclin-1 expression in pancreatic ductal adenocarcinoma

Beclin-1 has emerged as an autophagy gene in a variety of human carcinomas. The aim of this study was to evaluate beclin-1 expression and to determine its prognostic significance in patients with pancreatic ductal adenocarcinoma (PDA).We performed immunohistochemical staining for beclin-1 in 63 cases of PDA. We investigated whether beclin-1 expression correlated with clinicopathologic characteristics and patient outcomes.Beclin-1 expression was absent in normal pancreatic islet cells, acinar cells, and ductal epithelial cells. In contrast, in PDA, beclin-1 showed positive immunoreactivity in 14 of 63 (22.2%) PDA cases, while the remaining 49 (77.8%) PDA cases exhibited negative beclin-1 expression. We found significant associations between increased beclin-1 expression and the absence of lymphatic invasion (P = 0.032) and low rate of distant metastasis (P = 0.001). Univariate analysis of survival showed that the median distant metastasis-free survival of beclin-1-negative PDA patients (10 months) was significantly shorter than that of beclin-1-positive PDA patients (21 months; P = 0.031).Our results suggest that increased beclin-1 expression plays a role in the inhibition of PDA progression.     

PD-L1 expression in pancreatic adenosquamous carcinoma: PD-L1 expression is limited to the squamous component

Aim

We examined the programmed death-ligand 1 (PD-L1) expression in surgically resected pancreatic adenosquamous carcinoma (PASC) samples. Furthermore, the detection rate was also assessed using biopsy cases obtained from endoscopic ultrasound-guided fine needle aspiration (EUS-FNA).

神经源性分化蛋白在胰腺癌中的表达和意义

目的 探讨神经源性分化蛋白(NeuroD)在胰腺外分泌癌中的表达及其意义.方法 应用组织芯片和免疫组织化学EnVision二步法研究NeuroD、增殖细胞核抗原(PCNA)、p53在127例胰腺外分泌癌中的表达情况,并在光镜下观察胰腺癌神经组织浸润、神经周围淋巴细胞套、癌旁慢性胰腺炎、胰周淋巴结转移情况.分析NeuroD的表达与上述其他指标及性别、年龄、肿瘤部位、肿瘤组织学类型及分化程度等的关系.结果 NeuroD、PCNA和p53蛋白在胰腺癌组织中的阳性率分别为64.6％(82/127)、57.5％ (73/127)和59.1％ (75/127),与菲癌组织[分别为10.5％ (8/76)、9.2％(7/76)和9.2％(7/76)]相比,差异有统计学意义(P＜0.01).NeuroD蛋白的表达与PCNA、p53蛋白的表达和胰腺癌肿瘤神经浸润之间有统计学相关性(P＜0.05),而与性别、年龄、肿瘤部位、肿瘤组织学类型及分化程度、癌旁慢性胰腺炎、神经周围淋巴细胞套和淋巴结转移均等无统计学相关性(P＞0.05).结论 NeuroD蛋白在胰腺癌中呈高表达,可能参与了胰腺癌的发生和进展,并且与胰腺癌的增殖、p53信号通路和肿瘤神经浸润之间密切相关.