明胶酶、CD44v6在结直肠癌中的表达及意义

目的探讨明胶酶在结直肠癌中的表达及其CD44v6在结直肠癌侵袭转移过程中的关系。方法采用明胶酶谱（gel zymography）和S-P免疫组化对50例结直肠癌癌组织和相应正常组织中的明胶酶、CD44v6进行检测。结果明胶酶在结直肠癌中的表达水平显著高于正常组织（P〈0．05）；明胶酶与结直肠癌的Duke’s分期呈同步的正相关（P〈0．05）；明胶酶在CD44v6强阳性组中表达量升高显著（P〈0．05）。结论明胶酶与结直肠癌的侵袭转移性有着密切的关系，不仅在癌细胞的酶解，而且在癌细胞的黏附、运动过程中有可能发挥着重要作用。     

Mortality association of enhanced CD44v6 expression is not mediated through occult lymphatic spread in stage II colorectal cancer

BACKGROUND AND AIMS: In the absence of other metastatic disease, the presence of lymph node metastasis remains the most important determinant of survival in colorectal cancer (CRC). Cluster designation 44 variant 6 (CD44v6) over-expression is associated with worse outcome in all stages of CRC. The CD44v6 is believed to confer metastatic potential through its facilitation of migration, extravasation and proliferation, although the specific means by which it conveys an adverse prognosis in CRC is unknown. The aim of the present study was to determine if CD44v6 over-expression in Stage II CRC subjects was associated with the presence of lymph node micrometastases. METHODS: We assessed tumour CD44v6 expression in 43 randomly sampled subjects who had resections for Stage II CRC between 1984 and 1991 by using immunohistochemistry. Micrometastases were sought in corresponding lymph node (LN) sections using keratin immunohistochemistry. RESULTS: There was a statistical trend between tumour CD44v6 over-expression and mortality (P = 0.09) and a significant relationship between LN cytokeratins and mortality (P = 0.01). There was no association between the detection of LN cytokeratins and tumour CD44v6 over-expression. CONCLUSION: We conclude that the adverse survival effect of CD44v6 over-expression is not mediated though lymphatic spread and postulate that it may therefore facilitate haematogenous metastasis.     

FAK, CD44v6, c-Met and EGFR in colorectal cancer parameters: tumour progression, metastasis, patient survival and receptor crosstalk

Purpose

Research for reliable and patient-specific markers in colorectal cancer (CRC) is based on solid evidence that staging alone is not informative enough. Employing four cellular receptors, we embarked to identify aggressive tumour behaviour and impact of surrogate marker expression on patient prognosis.

Methods

One-hundred eighty-three CRC patients were enrolled in our investigation that focused on an array of biological markers, namely epidermal growth factor receptor (EGFR), c-Met, focal adhesion kinase (FAK) and CD44v6. Tissue samples, clinicopathological data and patient’s follow-up information were collected, and immunohistochemical assays evaluated the levels of the aforementioned molecules. All available data were correlated with tumour grade, stage, patient age, gender and survival.

Results

Expression of all receptors correlated closely with tumour stage (P?<?0.01) exhibiting a connection with cancer’s invasiveness and progress. Survival also proved to depend significantly on molecular expression (log-rank test for Kaplan–Meier; EGFR P?=?0.030, c-Met P?=?0.050, FAK P?<?0.001, CD44v6 P?<?0.001). Stage, FAK and CD44v6 emerged as independent predictors of survival in a stepwise regression analysis (FAK P?=?0.001 Exp(B)?=?2.517, 95 % confidence interval (CI)?=?1.704–5.831 and CD44v6 P?=?0.005, Exp(B)?=?2.299, 95 % CI?=?1.287–4.110). T-stage, nodal metastasis, all metastatic types (N/M) and size correlated with at least one of the receptors or their co-expression. Notably, increased staining for each receptor was followed by statistically significant expression elevation of at least one of the other markers.

Conclusions

Our results suggest that the selected cellular receptors are suitable for use as biomarkers of survival and tumour progression in CRC. Furthermore, we provide additional evidence for receptor interaction, properly clarifying their importance, which could potentially lead to more effective anti-CRC regimens.     

骨桥蛋白及 CD44 v6在肺腺癌侵袭转移中的作用

目的：探讨骨桥蛋白（ OPN）及其受体CD44v6在肺腺癌侵袭转移中的作用。方法应用免疫组化方法检测129例癌旁组织、肺原位腺癌（AIS）、伏壁样生长为主型腺癌（LPA）患者手术切除标本中OPN、CD44v6的表达情况。检测H358细胞（原位癌细胞系）和A549细胞中OPN、CD44v6蛋白及mRNA的表达,Transwell 试验观察OPN、CD44 v6对H358细胞、A549细胞侵袭力的影响。结果 OPN、CD44 v6在LPA组织中的表达阳性率显著高于AIS及癌旁组织,AIS高于癌旁组织,OPN与CD44v6的表达呈显著正相关（ P＜0．05）。 OPN、CD44v6及OPN mR-NA、CD44v6 mRNA在A549细胞中的表达水平高于H358细胞。 Transwell试验提示OPN对A549细胞的趋化作用高于H358细胞。使用OPN抗体阻断OPN对两种细胞的趋化作用,两种细胞穿过数均减少;阻断CD44 v6受体通道后,OPN对两种细胞的趋化作用明显减弱。结论 OPN-CD44 v6复合体对肺腺癌的侵袭能力具有重要作用,OPN有望成为评估肺腺癌进展及预测肿瘤转移潜能的指标。     

Expression of CD44v6 has no prognostic value in patients with colorectal cancer

BACKGROUND AND AIMS: In colorectal cancer patients' mortality is largely influenced by spreading of tumour cells from the primary tumour site and subsequent metastasis formation. CD44 is an adhesion molecule and represents a highly variable family of isoforms. The isoform CD44v6 has been associated with metastatic spread and poor prognosis in animal models and several human cancers. Results of immunohistological studies in primary colorectal cancer are mostly retrospective and contradictory. The aim of our prospective study was to assess the controversial role of CD44v6 as a prognostic factor in colorectal cancer. METHODS: In 93 patients we analysed tumour CD44v6 expression in prospectively sampled stage I-IV colorectal adenocarcinomas using RT-PCR and Southern blotting. The prognostic value of the CD44v6 expression was assessed using univariate and multivariate analysis. RESULTS: CD44v6 expression was found in 47 % of the cases. CD44v6 expression failed to show any association with the clinical or histological variables examined. CD44v6 expression did not correlate with survival in long-term follow-up. The most important prognostic factor in this cohort was tumour stage. CONCLUSIONS: Changes in CD446 expression level do not predict tumour spread or patient survival in colorectal cancer.     

Expression of CD44 variant isoforms CD44v3 and CD44v6 is increased on T cells from patients with systemic lupus erythematosus and is correlated with disease activity

Objective

To quantify the expression of CD44 and variant isoforms CD44v3 and CD44v6 on T cells from patients with systemic lupus erythematosus (SLE), and to assess correlations of the level of expression of these molecules with disease manifestations.

Methods

Information on clinical and demographic characteristics was collected, and blood samples were obtained from 72 patients with SLE and 32 healthy control subjects matched to the patients by sex, race, and age. Expression of CD44 and variants CD44v3 and v6 on T cell subsets was determined by flow cytometry, and Pearson's correlations of their expression levels with clinical variables, SLE Disease Activity Index (SLEDAI) scores, and presence of lupus nephritis were determined. Wilcoxon's rank sum tests and conditional multivariable regression analyses were applied to identify differences in the expression of CD44 between patients with SLE and healthy controls.

Results

Expression of CD44 was higher on CD4+ and CD8+ T cells from SLE patients compared with controls (P ≤ 0.03). Expression of CD44v3 and CD44v6 was also higher on total T cells and CD4+ and CD8+ T cells from SLE patients compared with controls (P ≤ 0.03). Cell surface levels of CD44v3 on total T cells, CD4+ T cells, and CD8+ T cells as well as cell surface expression of CD44v6 on total T cells and CD4+ T cells were correlated with the SLEDAI score (P < 0.05). The presence of lupus nephritis was associated with the expression of CD44v6 on total T cells, CD4+ T cells, and CD4−CD8− T cells (P < 0.05). Positivity for anti–double‐stranded DNA antibodies was associated with the expression levels of CD44v6 on T cells (P < 0.05).

Conclusion

These results indicate that expression levels of CD44v3 and CD44v6 on T cells may represent useful biomarkers of SLE activity.

     

结直肠癌组织中CD44V3,v6蛋白的表达意义

目的研究CD44v3,v6蛋白在结直肠癌组织中的表达及其预后意义.方法应用EnvisionTM免疫组化法,回顾性分析121例结直肠癌石蜡组织CD44v3,v6的表达.患者中位随访时间为67.77mo.结果CD44v3,v6在结直肠癌中的阳性率分别为60.3%和57.9%.CD44v3的阳性表达与患者肿瘤部位,淋巴结转移状况、远处转移与否、临床分期密切相关(P＜0.05,Spearman等级相关检验).CD44v6的阳性表达与患者性别、淋巴结转移状况、远处转移与否、临床分期密切相关(P＜0.05,Spearman等级相关检验).Kaplan-Meier生存曲线,Log-rank检验单因素生存分析结果显示,CD44v3阴性、阳性患者的五年生存率分别为81.25%,60.27%,两者之间存在显著性差异(P=0.035);CD44v6阴性、阳性患者的5 a生存率分别为80.39%,60.00%,两者之间亦存在显著性差异(P=0.0299).Cox模型多因素生存分析结果显示,CD44v3表达是影响结直肠癌预后的独立因素.结论CD44v3,v6蛋白与结直肠癌转移、预后相关,CD44v3是影响结直肠癌预后的独立因素.     

老年大肠癌患者外周血CD44v5、CD44v6表达及化疗对其影响

目的　研究老年大肠癌患者外周血CD44v5、CD44v6表达水平与血行转移的关系及化疗对其影响。方法　应用流式细胞仪检测 49例老年大肠癌患者手术前外周血CD44变异体 (CD44v5、CD44v6)表达水平 ,与健康老年人及非老年大肠癌患者进行对照 ,并在应用化疗后观察黏附分子表达水平的变化。结果　老年大肠癌组外周血CD44v5、CD44v6的表达水平分别为 1 0 71± 5 44、1 4 62± 6 34 ,明显高于健康对照组 (P <0 0 1 ) ,与非老年大肠癌组比较无显著性差异 (P >0 0 5)。CD44v5、CD44v6的表达水平与老年大肠癌患者肿瘤大小、肿瘤浸润深度及淋巴结转移相关 ,老年大肠癌组肿瘤≥ 5cm、肿瘤浸润深度≥深肌层、淋巴结转移阳性患者外周血CD44v5、CD44v6的表达水平分别高于肿瘤 <5cm、肿瘤浸润深度 <深肌层、淋巴结转移阴性患者 (P <0 0 1、P <0 0 5、P <0 0 1 )。老年大肠癌患者化疗后外周血CD44v5、CD44v6表达水平较化疗前明显下降 (P<0 0 1 )。结论　黏附分子具有介导老年人大肠癌细胞血行转移的作用 ,化疗可降低老年大肠癌患者外周血CD44v5、CD44v6的表达水平 ,具有抗黏附和肿瘤转移作用     

CD44v3和CD44v6在溃疡性结肠炎与其他类型结肠炎中的表达

目的比较CD44v3和CD44v6在溃疡性结肠炎(UC)与对照组(包括感染性结肠炎、阿米巴痢疾、血吸虫性结肠炎及克罗恩病等其他类型结肠炎)中表达的差异.方法免疫组化SP法检测两组患者结肠粘膜CD44v3和CD44v6的表达.结果 UC组患者CD44v3和CD44v6表达阳性率分别为68.8%和56.3%,与对照组比较有显著性增高(P<0.05).UC组与感染性结肠炎、阿米巴痢疾、血吸虫性结肠炎病及克罗恩病分别比较差异仍有显著性(P<0.05).结论 CD44v3和CD44v6的检测有助于鉴别UC和感染性结肠炎等其他类型结肠炎.     

Expression of human chorionic gonadotropin, CD44v6 and CD44v4/5 in esophageal squamous cell carcinoma

AIM: To study the relationship between the expression of human chorionic gonadotropin (HCG), CD44v6, CD44v4/5 and the infiltration, metastasis of esophageal squamous cell carcinoma. METHODS: By labeled streptavidin-biotin technique, the expressions of HCG, CD44v6, and CD44v4/5 in 42 patients with esophageal squamous cell carcinoma were examined. RESULTS: The positive rate of HCG expression in patients with lymph node metastasis was 85.71% (18/21), higher than that (57.14%, 12/21) in those without lymph node metastasis (P<0.05). The positive rate of CD44v6 expression was 71.43% (15/21) in lymph node metastasis group, and 38.09% (8/21) in non-metastasis group; there was a significant difference between the two groups (P<0.05). The positive rate of CD44v4/5 expression was 76.19% (16/21) in lymph node metastasis group, and 42.86% (9/21) in non-metastasis group; there was also a significant difference between them (P<0.05). From grade Ⅰ to grade Ⅲ in differentiation, the positive rate of HCG expression was 84.62% (11/13), 70.59% (12/17) and 58.33% (7/12), respectively; there was no significant difference among them (P<0.05). The positive rate of CD44v6 expression in grades Ⅰ-Ⅲ of cancer tissues was 76.92% (10/13), 52.94% (9/17), and 33.33% (4/12) respectively; there was no significant difference among them. The positive rate of CD44v4/5 expression in grades Ⅰ-Ⅲ of cancer tissues was 69.23% (9/13), 64.71% (11/17), and 41.67% (5/12) respectively; there was no significant difference among the three groups. There was no correlation between the positive rates of HCG and CD44v6, CD44v4/5 expression. Cancer cells in carcinomatous emboli and those infiltrating into vascular wall strongly expressed HCG, CD44v6, and CD44v4/5. CONCLUSION: Expression of HCG, CD44v6, and CD44v4/5 in esophageal squamous cell carcinoma is related to its infiltration and metastasis. HCG, CD44v6, and CD44v4/5 have different effects on the infiltration and metastasis of esophageal squamous cell carcinoma.     

Expression and clinical significance of CD44V5 and CD44V6 in resectable colorectal cancer

Purpose This study was conducted to evaluate the prognostic significance of CD44v5 and CD44v6 in resectable colorectal cancer.Materials and methods Membranous CD44v5 and CD44v6 levels were measured by an immunoenzymatic assay in tumors and surrounding mucosal samples obtained from 105 patients with resectable colorectal carcinomas.Results There were no significant differences of CD44v5 levels between tumors [median: 3.2 (range: 0.9–83.5) ng/mg protein) and surrounding mucosal samples (3 (3–146.2) ng/mg protein]. However, tumor samples showed significantly higher CD44v6 levels [19.5 (2.2–562.9) ng/mg protein] than mucosal samples [5 (5–230) ng/mg protein] (P=0.0001). Patients with higher CD44v5 or CD44v6 content in tumor samples had a considerably shorter relapse-free survival (P<0.05, for both). Patients with a higher CD44v6 content also had a shorter relapse-free and overall survival in the multivariate analysis (P<0.05).Conclusion The results of this study suggest a role of CD44v5 and CD44v6 in colorectal cancer progression. Membranous CD44v levels in primary tumors, measured by immunoenzymatic assay, may contribute to a more precise prognostic estimation in patients with resectable colorectal cancer.Supported by grants from ISCIII Red de Centros de Cancer RTICCC (C03/10) and Obra Social Cajastur     

Expression and significance of CD44s, CD44v6, and nm23 mRNA in human cancer

AIM: To investigate the relationship between the expression levels of nm23 mRNA, CD44s, and CD44v6,and oncogenesis, development and metastasis of human gastric adenocarcinoma, colorectal adenocarcinoma,intraductal carcinoma of breast, and lung cancer.METHODS: Using tissue microarray by immuhistochemical (IHC) staining and in situ hybri-dization (ISH), we examined the expression levels of nm23mRNA, CD44s, and CD44v6 in 62 specimens of human gastric adenocarcinoma and 62 specimens of colorectal adenocarcinoma; the expression of CD44s and CD44v6in 120 specimens of intraductal carcinoma of breast and 20 specimens of normal breast tissue; the expression of nm23 mRNA in 72 specimens of human lung cancer and 23 specimens of normal tissue adjacent to cancer.RESULTS: The expression of nm23 mRNA in the tissues of gastric and colorectal adenocarcinoma was not significantly different from that in the normal tissues adjacent to cancer (P＞0.05), and was not associated with the invasion of tumor and the pathology grade of adenocarcinoma (P＞0.05). However, the expression of nm23 mRNA was correlated negatively to the lymph node metastasis of gastric and colorectal adenocarcinoma (r = -0.49, P＜0.01; r = -4.93, P＜0.01). The expression of CD44s in the tissues of gastric and colorectal adenocarcinoma was significantly different from that in the normal tissues adjacent to cancer (P＜0.05;P＜0.01). CD44v6 was expressed in the tissues of gastric and colorectal adenocarcinoma only, the expression of CD44v6 was significantly associated with the lymph node metastasis, invasion and pathological grade of the tumor (r = 0.47, P＜0.01; r = 5.04, P＜0.01). CD44sand CD44v6 were expressed in intraductal carcinoma of breast, the expression of CD44s and CD44v6 was significantly associated with lymph node metastases and invasion (P＜0.01). However, neither of them was expressed in the normal breast tissue. In addition, the expression of CD44v6 was closely related to the degree of cell differentiation of intraductal carcinoma of breast (x2= 5.68, P＜0.05). The expressional level of nm23mRNA was closely related to the degree of cell differentiation (P＜0.05) and lymph node metastasis (P＜0.01), but the expression of nm23 gene was not related to sex, age, and type of histological classification (P＞0.05).CONCLUSION: Patients with overexpression of CD44s and CD44v6 and low expression of nm23 mRNA have a higher lymph node metastatic rate and invasion. In addition, overexpression of CD44v6 is closely related to the degree of cell differentiation. Detection of the three genes is able to provide a reliable index to evaluate the invasion and metastasis of tumor cells.     

Low serum levels of CD44, CD44v6, and neopterin indicate immune dysfunction in chronic pancreatitis

INTRODUCTION: In autoimmune diseases, malignancies, and inflammatory conditions, a correlation of serum levels of CD44, interleukin-2 receptor (IL-2r), and neopterin with disease activity could be shown. AIMS: To assess the immune parameters in chronic pancreatitis in correlation to clinical data to evaluate the potential role of immune dysfunction as a risk factor. METHODOLOGY: Levels of IL-2r, sCD44, sCD44v6, and neopterin were measured using the enzyme-linked immunosorbent assay in 63 patients with chronic pancreatitis who underwent surgery between 1992 and 1995 in our institution. Clinical data were evaluated prospectively before surgery, and a follow-up investigation was conducted in 1997. RESULTS: Mean serum levels of CD44, CD44v6, and neopterin were significantly lower in patients with chronic pancreatitis compared with the control group. The mean level of IL-2r was also lower in chronic pancreatitis, but this difference was not significant. However, no influence of immunosuppressive factors such as alcohol consumption, cigarette smoking, or diabetes could be detected on the levels of IL-2r, CD44, CD44v6, and neopterin. CONCLUSION: In accordance with other diseases of reduced immunoreactivity, depressed serum levels of biomarkers in chronic pancreatitis are caused by reduced T-lymphocyte and macrophage activation. By ruling out a significant influence of concomitant immunosuppressive factors, we conclude that the inflammatory process itself is the source of the depressed immune function, which might be restored by surgical resection.