Expression of CD44 variant isoforms and association to the benign form of endocrine pancreatic tumours

Background:The expression of CD44 and its isoforms have beenshown in many neoplastic tissues to serve as prognostic indicators, therefore,the feasibility of using these as prognostic markers in endocrine pancreatictumour patients was examined. Patients and methods:Immunohistochemistry (IHC) was performed on26 tumour samples (5 gastrinomas, 3 glucagonomas, 10 non-functioning tumours,6 insulinomas, 2 mixed insulinoma and glucagonomas) with monoclonal antibodiesagainst CD44s (standard form) and variant isoforms (v4, v5, v6, v7,v7–8, v9, v10). Staining was correlated to the tumour proliferation,malignancy, metastasis and patients survival. Results:There was variable expression of CD44s. All tumoursshowed complex expression of many isoforms. CD44v6 and CD44v9 were downregulated in malignant tumours. There was statistical significance of CD44v6expression in benign tumours (P < 0.05) compared to malignanttumours and near significance in CD44v9 expression (P = 0.0574).Survival of the patients with CD44v6 positive staining was higher than thosewho were negative (P = 0.0822). Moreover, the expression was wellcorrelated to the patients without any distant metastases (CD44v6, p<0.001;CD44v9, P < 0.01). Tumour proliferation (Ki67 index) correlateddirectly to the malignancy (P < 0.05) and there was inversecorrelation between Ki67 index and CD44v6 (P < 0.05) as well asv9 (P < 0.05). Conclusions:Endocrine pancreatic tumours express CD44s andisoforms differentially. Expression of the two isoforms of CD44, namely v6 andv9 seem to be related more to benign form of the tumour and could serve as apredictor of good prognosis.     

CD44v6 is not a prognostic factor in primary breast cancer

Background: CD44 is an adhesion molecule and represents a highly variable family of isoforms. The isoform CD44v6 has been associated with metastasis formation and poor prognosis in animal models and human colon cancer. Results of studies in primary breast cancer are relatively small and contradictory.Patients and methods: The immunohistochemical expression of CD44v6 was studied in a series of 338 patients with primary breast tumours, uniformly staged and treated in a single center with a long median follow-up of 128 months. The prognostic significance of CD44v6 as well as the correlation with several clinicopathological features were analysed.Results: Two hundred nineteen of 338 (64.8%) of the breast cancers were CD44v6-positive (>5% of tumour cells with positive staining). CD44v6 expression had no value for prognosticating disease-free or overall survival at this or any other cut-off point.Conclusion: CD44v6 expression is not a prognostic factor in primary breast cancer.     

Clinicopathological Significance of Expression of CD44 Variants in Head and Neck Squamous Cell Carcinoma

Splice variants of the cell surface glycoprotein CD44 have been reported to be associated with the progression of various human tumors. The aim of this study is to determine the correlation between the expression of CD44 isoforms, especially CD44 variant 2 (CD44v2), and the clinicopathological features of head and neck squamous cell carcinomas (HNSCCs). The expression of CD44 isoforms was evaluated immunohistochemically in paraffin-embedded tissues from 89 primary lesions, using monoclonal antibodies against CD44 standard (CD44st), CD44 variant 6 (CD44v6) and CD44v2. Cancer tissues from 89 (100%), 85 (95.5%) and 59 (66.3%) patients showed positive immunoreactivity for CD44st, CD44v6 and CD44v2, respectively. A significant correlation was observed between the down-regulation of CD44v2 and poorer differentiation of the tumor cells ( P =0.02). We could not find any significant correlation between the expression of CD44v2 and T stage or N stage (lymph node status). However, the rate of positive cervical lymph node metastasis tended to increase with reduced expression of CD44v2 ( P =0.08). Down-regulation of CD44v2 expression was correlated with shorter overall survival ( P =0.01). Furthermore, Cox's multivariate analysis revealed that only CD44v2 expression and lymph node status were independent prognostic factors. These findings suggest that down-regulation of CD44v2 expression may be one of the biological markers for the degree of malignancy in HNSCCs.     

E-cadherin expression as predictive marker of proximal resection line involvement for advanced carcinoma of the gastric cardia

AIMS: Total gastrectomy for gastric cardia tumours harbours a high risk of proximal resection line (PRL) involvement. The adhesion markers CD44v6 and E-cadherin were evaluated as predictive factors for PRL involvement independent of tumour stage. METHODS: Forty-nine gastrectomy specimens for gastric cardia carcinoma (type II) were evaluated for stage, resection margins, and CD44v6 and E-cadherin immunohistochemistry. RESULTS: PRL involvement was microscopically recognized in 49% of specimens. CD44v6 expression was found in 84% of intestinal tumours, and in 56% of diffuse/mixed tumours (p=0.045). In the group of resections performed with curative intent, the proximal extension of the resection (margin) was significantly shorter in E-cadherin negative tumours than in E-cadherin positive tumours (p=0.029). Histological type and stage of the tumour, lymph node metastases, and absence of E-cadherin expression, but not the presence of CD44v6 correlated with PRL involvement. Only the absence of E-cadherin expression appeared to be a significant predictor of PRL involvement, independent of tumour stage. Survival for patients with PRL involvement was shorter than that for patients after R0 resection (p=0.07). Stage was the only independent prognostic factor emerging from multivariate survival analysis (p=0.002). CONCLUSIONS: When curative resection is intended in type II cardiac cancer patients, an oesophageal resection and gastric tube reconstruction should be considered, especially for a tumour without E-cadherin expression.     

E-cadherin, CD44s and CD44v6 correlate with tumour differentiation in colorectal cancer

Cell adhesion molecules (CAMs) are cell surface glycoproteins that are important in cell-cell and cell-matrix interactions and play an important role in cell growth and differentiation. We examined immunohistochemically CD44s, CD44v6 and E-cadherin expression in 86 formalin-fixed, paraffin-embedded primary tumours and 5 metastases. Lower levels of CD44s, CD44v6 and membranous E-cadherin expression were significantly associated with higher tumour grade (p=0.022, p=0.016 and p= 0.041, respectively). Moreover, CD44v6 and membranous E-cadherin expression were correlated with the depth of primary tumour invasion (p=0.030 and p=0.020, respectively), and increased expression of CD44v6 and decreased membranous E-cadherin expression were associated with increased primary tumour invasion. The results suggest that these CAMs are associated with tumour differentiation and invasion in locally advanced and metastatic colorectal carcinoma.     

IMMUNOCYTOCHEMICAL CHARACTERIZATION OF CD44 MOLECULES EXPRESSED IN HUMAN BRAIN METASTASES

ＩＭＭＵＮＯＣＹＴＯＣＨＥＭＩＣＡＬＣＨＡＲＡＣＴＥＲＩＺＡＴＩＯＮＯＦＣＤ４４ＭＯＬＥＣＵＬＥＳＥＸＰＲＥＳＳＥＤＩＮＨＵＭＡＮＢＲＡＩＮＭＥＴＡＳＴＡＳＥＳＬｉＨｏｎｇ李宏；ＬｉｕＪｉａ刘佳；ＭａｒｔｉｎＨｏｆｍａｎｎ；Ｍａｒｉａｅ－Ｆｒａｎｃｅ...     

CD44v6与可溶性CD44v6在胃癌中的表达及与预后的关系

目的 探讨CD44v6与可溶性CD44v6(sCD44v6)在胃癌中的表达及与胃癌生物学行为和预后的关系.方法 应用免疫组化S-P法检测103例胃癌组织和10例正常胃黏膜标本的CD44v6表达,并对患者进行术后随访,随访期为3～91个月;应用酶联免疫吸附(ELISA)法检测86例胃癌、30例胃溃疡患者和30例健康人外周血血清中sCD44v6的浓度,并对患者进行术后随访,随访期为1～91个月.结果 (1)CD44v6在正常胃黏膜中不表达,而在胃癌中的表达率为60.2%;CD44v6的表达与淋巴结转移、TNM分期、脉管是否存在癌栓以及Borrmann分型有关(P＜0.05),而与胃癌浸润深度、是否远处转移、分化程度以及患者的生存率无关(P＞0.05);9例伴有肝转移的胃癌患者中,有7例CD44v6为强阳性表达.(2)胃癌患者外周血血清中sCD44v6的浓度明显高于胃溃疡组和健康对照组(P＜0.01);胃癌根治术后患者血液中sCD44v6的浓度比术前明显下降(P＜0.01),而姑息术后患者的浓度变化却不明显(P＞0.05);sCD44v6的浓度与胃癌浸润深度、淋巴结转移、是否远处转移、脉管是否存在癌栓、Borrmann分型以及分化程度无关;sCD44v6浓度较高组的生存率较sCD44v6浓度较低组高,且差异有统计学意义(P=0.0281),但经Cox回归分析显示,sCD44v6的浓度与生存期有关(P=0.415),而手术方式却与生存期有关(P=0.000).(3)胃癌组织中CD44v6的表达与外周血血清中sCD44v6的表达无明显平行关系(P＞0.05).结论 CD44v6的表达对评估胃癌生物学行为有参考价值,但是否与预后有关仍需探讨;sCD44v6的表达对胃癌辅助诊断、手术彻底性判定及生物学行为评估有一定的意义;CD44v6与sCD44v6的表达无明显平行关系.     

CD44v6、p27表达与大肠癌临床病理学因素的关系

目的探讨CD44v6和p27在大肠癌中的表达及其临床意义。方法对50例手术切除的大肠癌新鲜标本及相应的正常大肠黏膜,应用流式细胞术（FCM）检测CD44v6的表达;同步制备上述病例大肠癌组织石蜡标本,采用免疫组织化学方法检测大肠癌组织中p27的表达,结合病理及临床资料进行统计学分析。结果CD44v6和p27在大肠癌组织及正常黏膜组织中表达存在明显差异（P〈0．01）,而且CD44v6和p27的表达与大肠癌的分化程度、有无淋巴结转移、有无远端器官转移、Duke分期及预后相关（P值均〈0．05）,CD44v6。的表达与大肠癌的浸润深度无明显相关（P〉0．05）,但p27的表达与大肠癌的浸润深度明显相关（P〈0．05）。结论CD44v6和p27的表达与大肠癌的临床病理学特征密切相关,两者在大肠癌组织中的表达呈一定的负相关,同时检测二者对判断大肠癌的恶性程度、复发转移潜能,评价患者的预后及术后选择合理的治疗方案具有一定的参考价值。     

Soluble CD44 v5 and v6 in serum of patients with breast cancer. Correlation with expression of CD44 v5 and v6 variants in primary tumors and location of distant metastasis

Primary breast cancers were shown to overexpress CD44 v5 and v6 at the plasma membrane. However, the clinical significance of this overexpression remains unclear. Overexpression of CD44 v5 and v6 in primary breast cancers was found to correlate with metastasis and poor prognosis by some investigators, yet this correlation could not be confirmed by others using different antibodies. In this study the influence of metastatic disease, the site of metastasis, and the amount of CD44 v5 and v6 expression in the primary tumor on serum levels of the soluble forms of CD44 v5 and v6 (sCD44 v5 and v6) in breast cancer patients was investigated. Soluble CD44 v5 and v6 serum levels were measured by enzyme linked immuno sorbent assay in a group of breast cancer patients who developed metastases in various organs and in another group of patients with single organ metastasis. For control, sCD44 v5 and v6 levels were measured in breast cancer patients who remained free of metastasis and in healthy blood donors. Expression of plasma membrane bound CD44 v5 and v6 in the primary tumors of the patients with metastasis in various organs was correlated to sCD44 v5 and v6 levels in serum. Furthermore the size of sCD44 v6 was analyzed by immunoblot using a monoclonal antibody directed against CD44 v6. When metastases were detected, sCD44 v5 and v6 serum levels were increased as compared to levels measured one month after tumor surgery in patients free of metastases (p = 0.0025 and p = 0.0004). Six of 19 and 6 of 20 patients had sCD44 v5 and v6 serum levels above a cut-off level of 85 and 275 ng/mL, respectively. In these cases expression of CD44 v5 and v6 in the primary cancers was also elevated. Low sCD44 v5 and v6 serum levels were associated with weak expression of CD44 v5 and v6 in the respective primary cancers. As shown by statistical analysis of sCD44 v5 and v6 levels in 57 patients with single organ metastases, elevated sCD44 v6 levels but not sCD44 v5 levels were associated with metastases in liver or bone (p = 0.0025). Immunoblot analysis of soluble CD44 proteins in serum revealed two CD44 v6 specific signals of approximately 120 and 170 kDa. Increased sCD44 v5 and v6 serum levels in patients with breast cancer were influenced by the amount of CD44 v5 and v6 expression in the primary tumor by the site of metastasis. Elevated sCD44 v6 serum levels were preferentially found in patients with metastases in liver or bone.     

CD44在肝细胞癌及癌周组织中的表达及其意义

 目的 研究CD44s及CD44v6蛋白在人肝细胞癌组织中表达及其意义。方法 应用免疫组化方法检测42例人肝细胞癌及其癌周组织与10例正常肝组织标本的CD44s及CD44v6表达情况。结果 CD44s及CD44v6在肝癌组织中表达明显高于癌周及正常肝组织，与门静脉内癌栓、包膜浸润及分化程度密切相关（分别为P＜0.01，P＜0.05，P＜0.05）。生存时间2年及2年以上组CD44s及CD44v6阳性表达分别为17.6 %（3／17），5.8 %（1／17），生存时间短于2年分别为52.6 %（10／19），42.2 %（8／19）（P＜0.05）。结论 肝细胞癌组织中CD44的表达与细胞分化、浸润转移密切相关，其阳性表达有助于判断预后。     

Expression of CD44 Variants and Its Association with Survival in Pancreatic Cancer

Since the CD44 variant 6(v6) molecule has been noted as a marker for tumor metastasis and prognosis in several tumors, we examined whether or not v6 is a useful marker for evaluating the prognosis of pancreatic cancer patients. In addition, we attempted to assess the clinicopathological implications for pancreatic cancer of the variant 2 (v2) isoform using a recently developed monoclonal antibody against a v2 epitope. The expression of CD44 variants was evaluated immunohistochemically in paraffin-embedded pancreatic cancer tissues from 42 patients who were confirmed surgically and histologically to have received curative resection. An indirect immunoperoxidase method was used with monoclonal antibodies against epitopes of the standard (CD44s) portion, v6 and v2. Protein expression data were evaluated statistically for any correlations with the length of survival or with histological parameters. The expression of CD44v6 and v2 was observed only in tumor cells, if at all. On the other hand, expression of total CD44 (including CD44v, as well as CD44s) was observed in both tumors and adjacent normal sites. Tumor tissue from 21 (50%) and 16 (38%) patients showed positive immunoreactivity with mAb 2F10 (anti-CD44v6) and mAb M23.6.1 (anti-CD44v2), respectively. The expression of CD44v6 and v2 was correlated with decreased overall survival ( P =0.0160 and P =0.0125, respectively). A significant correlation was obtained between CD44v2 peptide expression and vessel invasion ( P =0.026). These results suggest that CD44v2 and CD44v6 may be useful markers for poor prognosis in curatively resected primary pancreatic cancer.     

Serum CD44 splice variants in cervical cancer patients

Aberrant expression of the cell adhesion molecule CD44 has been detected in human tumours and has been shown to be associated with metastasis and poor prognosis in human malignancies. We evaluated serum levels of different soluble CD44 molecules (CD44 standard form and CD44 splice variants v5 and v6) in cervical cancer patients stage IB to IIIB. Two-hundred three serum samples were analysed. Serum levels of CD44st and CD44v5 showed no significant correlation with the presence or absence of cervical cancer. The splice variant CD44v6 showed a mean concentration of 227.3 ± 90.9 (minimum 71.4, maximum 543.9) ng/ml when tumour was present and a mean concentration of 198.7 ± 135.4 (minimum 67.2, maximum 696.3) ng/ml in cases of complete remission (P-value = 0.0001). However, in this preliminary study the sensitivity/specificity characteristic of CD44v6 was poor.     

CD44v3 and v6 variant isoform expression correlates with poor prognosis in early-stage vulvar cancer.

Expression of alternatively spliced CD44 isoforms has been reported to correlate with poor prognosis in human squamous cell cancers, i.e. squamous cell cancer of the lung and cervix. The aim of this study was to evaluate whether CD44 isoform expression is a prognostic factor in early-stage squamous cell cancer of the vulva. Seventy cases of squamous cell carcinoma of the vulva International Federation of Gynaecology and Obstetrics (FIGO) stage I were examined immunohistochemically for expression of CD44 isoforms. We used four different variant exon sequence-specific murine monoclonal antibodies to epitopes encoded by exons v3, v5, v6 and v7-8 of human variant CD44. The correlation of CD44 expression with histological grade and disease-free and overall survival was investigated. CD44 isoforms CD44v3, CD44v5, CD44v6 and CD44v7-8 were detected in 28% (20/70), 47% (33/70), 33% (23/70) and 17% (12/70) of the tumour samples respectively. Patients suffering from tumours expressing CD44v6 had a poorer relapse-free (log-rank test, P = 0.02) and overall survival (log-rank test, P = 0.03). Likewise, patients suffering from tumours expressing CD44v3 had a poorer relapse-free (log-rank test, P = 0.04) and overall survival (log-rank test, P = 0.01). Expression of CD44v5 and CD44v7-8 did not compromise the patients'' outcome. Histological grade did not correlate with CD44 isoform expression. Immunohistochemically detected expression of CD44 isoforms containing variant exon v6 or v3 is correlated with a poor relapse-free and overall survival in FIGO stage I vulvar cancer patients.     

CD44v6在乳腺浸润性导管癌组织中的表达及其临床意义

背景与目的:近年研究表明,CD44v6与乳腺癌的发生、侵袭和转移密切相关,但其与乳腺癌预后关系的报道结果并不一致。本研究探讨CD44v6在乳腺癌和乳腺癌旁非癌组织中的表达及其与乳腺癌临床病理因素的关系以及对乳腺癌患者预后的预测意义。方法:采用免疫组织化学SP法检测84例乳腺浸润性导管癌和20例癌旁非癌乳腺组织中CD44v6的表达。采用SPSS10.0软件统计分析CD44v6表达与各病理因素的关系,采用Cox比例风险模型分析影响预后的因素。结果:CD44v6在乳腺浸润性导管癌上皮细胞的表达(78.6%)明显高于癌旁非癌乳腺组织上皮(5.0%),差异有显著性(P<0.05);CD44v6的表达与乳腺癌的TNM分期、肿瘤大小、淋巴结转移状况密切相关;中位随访时间为60个月,CD44v6阴性组的总体生存情况优于CD44v6阳性组,有显著性差异(P<0.05),而且随着CD44v6表达的增强,总体生存曲线有逐渐下降的趋势。Cox比例风险模型多因素的预后分析结果显示,ER、TNM分期、CD44v6均是影响预后的独立因素(P<0.05)。结论:CD44v6在乳腺癌组织中的表达明显升高;CD44v6的表达与乳腺癌患者的TNM分期、肿瘤大小、淋巴结转移呈正相关;CD44v6表达升高可作为预测乳腺癌预后的独立指标之一。     

食管癌组织中上皮钙黏素、CD44v6和血管内皮生长因子C 表达与淋巴结转移的关系

 目的　探讨上皮钙黏素（E-cad）、CD44v6和血管内皮生长因子C（VEGF-C）在食管癌组织中的表达及其与淋巴结转移的关系。方法　应用免疫组织化学EnVision两步法,检测56例食管鳞状细胞癌（鳞癌）组织（区域淋巴结转移组32例,非转移组24例）和12例食管切缘正常黏膜组织中E-cad、CD44v6和VEGF-C的表达情况。结果　E-cad在56例食管鳞癌组织中的阳性表达率为55.4 %（31／56）,显著低于食管切缘正常黏膜组织的100.0 %（12／12）（P＜0.05）;CD44v6和VEGF-C在食管鳞癌组织中的阳性表达率分别为71.4 %（40／56）和62.5 %（35／56）,显著高于食管切缘正常黏膜组织的25.0 %（3／12）和16.7 %（2／12）（P＜0.05）;E-cad在区域淋巴结转移组阳性表达率为31.3 %（10／32）,明显低于非转移组的87.5 %（21／24）（P＜0.05）;CD44v6和VEGF-C在区域淋巴结转移组阳性表达率分别为84.4 %（27／32）和75.0 %（24／32）明显高于非转移组54.2 %（13／24）和45.8 %（11／24）（P＜0.05）;E-cad表达与食管癌淋巴结转移呈负相关,CD44v6和VEGF-C表达与淋巴结转移呈正相关,差异有统计学意义（P＜0.05）。结论　E-cad、CD44v6、VEGF-C在食管鳞癌淋巴结转移过程中起重要作用,联合检测有助于判断食管癌的生物学行为和预后。     

CD44v6、P27蛋白在乳腺癌组织中的表达及意义

目的 探讨乳腺癌中CD44v6、p27蛋白表达的意义。方法 应用免疫组化S-P法检测65例乳腺癌中CD44v6、p27蛋白表达。结果 65例乳腺癌中CD44v6、p27蛋白表达总阳性率分别为58.4％、38.4％。CD44v6蛋白阳性表达率与乳腺癌组织学类型、腋淋巴结转移呈正相关(P<0.05,P<0.01),与预后呈负相关(P<0.01),与组织学分级无明显相关性(P>0.05);p27蛋白阳性表达与乳腺癌组织学类型、组织学分级及腋淋巴结转移呈负相关(P<0.05),与预后呈正相关(P<0.05)。结论 CD44v6、p27蛋白表达与乳腺癌浸润、转移及预后显著相关,检测乳腺癌中CD44v6、p27蛋白表达水平,对判断乳腺癌恶性程度、复发转移潜能,评估患者预后及术后选择合理治疗方案有一定参考价值。     

Tiam1,CD44v6和Ecadherin表达与胃癌组织病理生物学行为的关系

目的探讨Tiam1、CD44v6和Ecadherin在胃癌组织中的表达及其与病理生物学行为的关系。方法应用RT-PCR和免疫组化SP法对56例胃癌和15例正常胃黏膜组织进行Tiam1、CD44v6和Ecadherin蛋白检测。结果与正常胃黏膜组织相比,Tiam1、CD44v6在胃癌组织中高表达（P=0.000,P=0.018）,Ecadherin在胃癌组织中低表达（P=0.001）;Tiam1、CD44v6、Ecadherin在胃癌组织中的表达与组织分化程度密切相关（P=0.022,P=0.016,P=0.018）;Spearman等级相关分析显示,Tiam1与CD44v6的表达呈正相关（γs=0.576,P〈0.01）,Tiam1与Ecadherin的表达呈负相关（γs=-0.428,P〈0.01）。结论Tiam1、CD44v6和Ecadherin的表达与胃癌的发生、发展和分化程度密切相关。     

CD44v6 和MRP 在卵巢癌中的表达

 目的 探讨CD44v6和MRP在上皮性卵巢癌组织中表达与化疗疗效和预后的关系。方法 采用免疫组织化学S-P法，检测10例正常卵巢组织、20例卵巢良性肿瘤和50例卵巢恶性肿瘤上皮性组织中CD44v6和MRP的表达。结果 ①CD44v6和MRP在卵巢恶性肿瘤中的表达明显高于在良性及正常组织中的表达（P〈0．05）；②CD44v6和MRP强阳性率与卵巢癌的临床分期、组织学分级和淋巴结转移有关（P〈0．05）；③卵巢癌中CD44v6和MRP的表达呈正相关（r=0．557，P〈0．05）；④CD44v6和MRP阳性表达的病例，化疗疗效和预后差。结论 检测卵巢癌中的CD44v6和MRP表达可反映肿瘤细胞的化疗疗效及预测预后。     

Expression of CD44 Isoforms in Infiltrating Lobular Carcinoma of the Breast

Expression of CD44 has been shown to correlate with the progression and prognosis of some malignant tumors. The aim of this study was to evaluate the expression of CD44 isoforms in infiltrating lobular carcinomas and analyse their potential as prognostic indicators. A panel of 39 tumors were examined for their expression of membranous and cytoplasmic CD44s, v3, v5, v6, v7 and v3-10 in the infiltrating cells, by immunohistochemical staining. The protein positive tumors showed membranous and/or cytoplasmic staining with all antibodies used except for CD44v7, which only displayed cytoplasmic staining. Cytoplasmic expression of CD44v3 (P=0.014) and membranous expression of v6 (P=0.039) were significantly associated with alveolar, classical/alveolar carcinomas and mucinous/alveolar carcinomas. Furthermore, in alveolar, classical/alveolar and mucinous/alveolar carcinomas, cytoplasmic staining of CD44v5 was correlated with lymph node negative patients (P=0.048), whereas membranous v5 was correlated with lymph positive patients (P=0.048). In classical, classical/trabecular and trabecular carcinomas expression of membranous CD44s was significantly correlated with lymph node status (P=0.042).