Thymidine phosphorylase and dihydropyrimidine dehydrogenase activity in colorectal carcinoma and patients prognosis

BACKGROUND AND AIMS: 5-Fluorouracil (5-FU) is a widely used for colorectal carcinoma. However, the therapeutic effect of 5-FU differs among patients. This difference may be based on the difference in sensitivity of carcinoma cells to 5-FU. The activities of thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) are reported to be correlated with cancer cell sensitivity against 5-FU in vitro. We evaluated whether TP and DPD are useful markers of tumor sensitivity for 5-FU in colorectal carcinomas. PATIENTS AND METHODS: We analyzed the TP and DPD in 189 patients with colorectal carcinoma using an enzyme-linked immunosorbent assay in relation to patients prognosis. RESULTS: The tumors' mean TP activity was significantly higher than that of noncancerous tissues (100 vs. 47 U/mg protein), but the tumors' mean DPD activity was significantly lower than that of noncancerous tissues (58 vs. 84 U/mg protein). Tumor TP, DPD, and TP/DPD values were not correlated with tumor location or histological types of tumors. Even tumor TP and TP/DPD values in Dukes' stage A tumors were lower than those of other stages; DPD values were not correlated with tumor stages. In 100 patients who underwent intravenous adjuvant 5-FU chemotherapy, prognosis was not correlated with tumor-TP, DPD, or TP/DPD values. Moreover, in 20 patients with synchronous liver metastasis who underwent postoperative 5-FU therapy through the hepatic artery, the survival times of patients was not correlated with tumor-TP, DPD, or TP/DPD values. CONCLUSIONS: These findings indicate that it is questionable to decide the indication of 5-FU chemotherapy according to tumor TP or DPD status in patients with colorectal cancer.     

检测微淋巴管及其密度和血管内皮生长因子C对结直肠癌的临床意义

目的探讨检测微淋巴管、微淋巴管密度(LMVD)和血管内皮生长因子C(VEGF-C)的表达对结直肠癌的临床意义。方法应用5′-核苷酸酶(5′-Nase)组织化学、SABC免疫组织化学(免疫组化)和RT-PCR检测80例结直肠癌组织和癌旁组织及30例结直肠正常组织的微淋巴管、LMVD和VEGF-C的表达,并随访、记录患者的临床病理参数和生存资料,分析其相关性。结果(1)结直肠癌、癌旁、正常结直肠组织的微淋巴管均被染成棕黄褐色。结直肠癌组织微淋巴管管腔封闭或无腔,为无功能淋巴管；癌旁组织微淋巴管丰富,管腔大,为功能性淋巴管。(2)癌旁组织LMVD为9．76±2．85,明显高于结直肠正常组织的5．49±．43(t=8．220,P＜0．01)；也高于癌组织的2．13±0．96(t= 15．118,P＜0．001)。(3)结直肠癌VEGF-C蛋白表达阳性率(48．8%)和相对表达量(1．09±1．20)明显高于正常结直肠组织(0和0),与VEGF-C mRNA表达一致；且VEGF-C表达与LMVD相关。(4) LMVD、VEGF-C表达与结直肠癌患者的年龄、性别、肿瘤部位大小、大体组织学类型无关(均P＞0．05)；与Dukes分期(P＜0．0001、P=0．0234)、淋巴结转移(P＜0．0001、P=0．0059)和生存期(P＜0．0001、P＜0．0001)密切相关。LMVD还与分化程度(P=0．0168)和肝肺血行转移(P=0．0088)相关。结论结直肠癌癌旁微淋巴管为功能性淋巴管；癌旁的功能性微淋巴管和增高的LMVD及肿瘤VEGF-C表达,可作为结直肠癌淋巴管生成的形态学特征、分子表型和判断结直肠癌患者淋巴转移及预后的重要指标。     

胸苷磷酸化酶和二氢嘧啶脱氢酶在原发性肝癌中的表达

目的探讨胸苷磷酸化酶（TP）和二氢嘧啶脱氢酶（DPD）在肝癌中的表达情况。方法选取肝癌手术切除标本20例，取肝癌和癌旁肝组织进行TP和DPD的免疫组织化学染色。运用图像分析系统对结果进行分析。结果TP在肝癌组织中表达的平均阳性单位值为26.696，在癌旁肝组织中表达的平均阳性单位值为30.996，差异有统计学意义（P〈0.05）。DPD在肝癌组织中的表达的平均阳性单位值为11.408，在癌旁肝组织中表达的平均阳性单位值为13.843，差异有统计学意义（P〈0.5）。肝癌组织中TP与DPD表达的平均阳性单位值的比值为4.447，在癌旁肝组织中TP与DPD表达的平均阳性单位值的比值为4.152。TP与DPD的比值（TP／DPD）在肝癌组织中较癌旁高，差异无统计学意义（P〉0．05）。结论肝癌和癌旁肝组织中均有较高水平的TP酶和DPD酶表达，TP与DPD的比值在肝癌组织中较癌旁高。     

HSP27 和claudin-10在结直肠癌中的表达及其临床意义

目的：探讨HSP27和claudin-10在结直肠癌中的表达及其临床意义。 方法：应用免疫组化法检测50例结直肠癌组织、25例结直肠腺瘤组织、50例正常结直肠黏膜组织中HSP27和claudin-10的表达,并分析结直肠癌中两者表达的关系以及与结直肠癌患者临床病理因素及预后的关系。 结果：在结直肠癌组织、结直肠腺瘤组织、正常结直肠黏膜中HSP27阳性表达率分别为54.0%、20.0%、16.0%,差异有统计学意义（P<0.001）;claudin-10阳性表达率72.0%、56.0%、54.0%,差异无统计学意义（P>0.05）;结直肠癌中HSP27与claudin-10的表达呈正相关（r=0.318,P=0.024）。单因素分析显示,HSP27的阳性表达率与与淋巴结转移有关（P<0.05）;claudin-10的阳性表达率与肿瘤直径、浸润深度及淋巴结转移有关（均P<0.05）。生存分析显示,HSP27和claudin-10阳性表达患者平均生存期明显低于各自的阴性表达患者（均P<0.05）。多因素分析显示,HSP27、claudin-10表达及淋巴结转移是影响结直肠癌患者预后的独立风险因素（均P<0.05）。 结论：HSP27在结直肠癌中组织中表达上调,并可能与claudin-10协同在结直肠癌转移中起重要作用。     

原发性肝癌中胸苷磷酸化酶和二氢嘧啶脱氢酶的表达及其临床意义

目的探讨胸苷磷酸化酶(TP)和二氢嘧啶脱氢酶(DPD)在肝癌及癌旁组织中的表达情况.方法本组自2004年2月至2005年10月选取肝癌手术切除标本24例,并取肝癌和癌旁肝组织进行TP和DPD的免疫组化染色,运用图像分析系统对结果进行分析.结果TP在肝癌和癌旁肝组织中表达的平均阳性单位值分别为25.76和31.96,两者差异显著(P＜0.05).DPD在肝癌和癌旁肝组织中的表达的平均阳性单位值分别为11.28和13.73,差异显著(P＜0.05).TP与DPD表达的平均阳性单位值的比值在肝癌和癌旁肝组织中分别为4.73和4.25.TP与DPD的比值(TP/DPD)在肝癌组织中较癌旁高,然而两者之间无显著性差异(P＞0.05).结论肝癌和癌旁肝组织中均有较高水平的TP酶和DPD酶表达,TP与DPD的比值在肝癌中比癌旁肝组织中有较高的趋势,表明在肝癌化疗中运用5-氟尿嘧啶前体药物有一定的理论基础.     

趋化因子受体CXCR4在结直肠癌组织中的表达及其与临床病理特征的关系

目的 观察趋化因子受体CXCR4在结直肠癌组织中的表达及其临床意义,探讨基质衍生因子SDF-1(CXCL12)-CXCR4生物学轴在结直肠癌侵袭转移中的作用.方法 免疫组织化学法检测53例结直肠癌组织及27例正常黏膜组织中CXCR4的表达及分布,并分析高表达CXCR4与结直肠癌患者临床病理特征的关系.反转录.聚合酶链反应(RT-PCR)、Western印迹方法检测27例结直肠癌肿瘤组织、正常黏膜及5例肝转移灶内CXCR4 mRNA表达及CXCR4蛋白表达水平.结果 53例结直肠癌组织中CXCR4表达阳性率为73.6%,高表达率(表达超过50%细胞)为45.3%;有淋巴结转移组CXCR4高表达率(65.4%)明显高于无淋巴结转移组(25.9%)(P＜0.01),并与肿瘤血管淋巴管浸润有关.随着临床分期的增加,肿瘤组织中CXCR4高表达率有升高趋势.但差异无统计学意义(P＞0.05).CXCR4的表达与患者肿瘤部位、大小、浸润深度无关(P＞0.05).27例新鲜结直肠癌组织中CXCR4 mRNA及蛋白表达水平明显高于相应的正常黏膜组织(P＜0.01);5例肝转移灶组织中CXCR4表达显著高于对应的原发灶(P＜0.01).结论 趋化因子受体CXCR4是一类参与结直肠癌病程进展的重要分子.CXCL12-CXCR4信号通路有望成为结直肠癌治疗的新靶点.     

生长激素受体在结直肠癌中的表达及其意义

目的研究生长激素受体(GHR)在结直肠癌中的表达情况,了解生长激素(GH)在结直肠癌发生、发展中的作用。方法应用免疫组织化学法(IHC)测定100例结直肠癌标本及其附近正常黏膜中GHR的表达情况,同时测定Ki-67表达。分析结直肠癌组织与正常黏膜GHR的表达及其与临床病理因素和肿瘤的关系。结果结直肠癌中GHR表达(81%)明显高于正常黏膜(68%),GHR的表达与肿瘤分化程度、病理分期、肿瘤大小相关(P=0.047、P=0.003和P=0.017),并与肿瘤增殖相关(P<0.001)。结论GHR在结直肠癌组织中有较高的阳性表达,提示GHR的表达是结直肠癌发生、发展过程中的重要事件,结直肠癌患者应用GH应慎重。     

Immunohistochemical analysis of thymidylate synthase, thymidine phosphorylase, and dihydropyrimidine dehydrogenase in rectal cancer (cUICC II/III): correlation with histopathologic tumor regression after 5-fluorouracil-based long-term neoadjuvant chemoradiotherapy

In locally advanced rectal cancer, neoadjuvant 5-fluorouracil (5-FU)-based long-term chemoradiotherapy leads to marked tumor reduction and decrease of local recurrence rate. Thymidylate synthase (TS), thymidine phosphorylase (TP), and dihydropyrimidine dehydrogenase (DPD) are known to be important biomarkers to predict tumor response to 5-FU-based therapy. The aim of this study was to examine the correlation between TS, TP, and DPD protein expression and histopathologic tumor regression after neoadjuvant chemoradiotherapy. The results were compared with the recently published mRNA data. Preoperative biopsies (n = 25) and resection specimens (n = 40) from patients with rectal carcinoma (clinical UICC stage II/III) receiving neoadjuvant 5-FU-based chemoradiotherapy were studied for TS, TP, and DPD protein expression by immunohistochemistry using three different scoring systems (intensity, pattern, intensity + pattern). Results were compared with histopathologic tumor regression. A significant correlation between protein expression and tumor response was only seen when both staining intensity and staining pattern were considered. With this method, a significant association was seen between high TS expression in tumor biopsies as well as resection specimens and nonresponse of the tumor to therapy (P = 0.04). Furthermore, low TP expression in the resection specimens was significantly associated with lack of response (P = 0.02). For DPD no significant correlations were found at all. In conclusion, these results suggest that immunohistochemistry like RT-PCR is a suitable method to determine the correlation between TS, TP, and DPD expression and histopathologic tumor regression. However, precise results can only be achieved if staining intensity as well as staining pattern within the tumors are evaluated.     

Thymidine phosphorylase and dihydropyrimidine dehydrogenase in bladder cancer

Thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD) levels in transitional cell carcinoma of the bladder resected from 38 patients were examined by ELISA. TP levels in high-grade and invasive cancer were significantly higher than those in low-grade and superficial cancer, respectively. No significant differences in the DPD levels were observed among grades and stages, but the DPD/TP ratio was significantly lower in grade 3 tumor than in grade 1. These results demonstrated that 5'-deoxy-5-fluorouridine seemed to be useful for managing patients with grade 3 cancer. The present study also suggested that we might be able to exclude cases of bladder cancer in which 5-fluorouracil group medicines would be inappropriate candidates in treatment options by measuring both TP and DPD levels in the tumor.     

促肝细胞再生磷酸酶-3基因在结直肠癌组织中的表达及其临床意义

目的检测促肝细胞再生磷酸酶-3(PRL-3)mRNA在结直肠癌(CRC)中的表达,探讨其与CRC发生发展以及侵袭转移的关系。方法半定量PCR测定46例CRC组织及其对应正常黏膜、6例结直肠腺瘤(CRA)及其对应正常黏膜以及18例淋巴结和肝转移灶中PRL-3mRNA相对表达水平,并分析其表达水平与临床病理学指标的关系。单链多态性分析法(PCR-SSCP)检测基因突变情况。结果46例CRC中PRL-3基因表达量较相应正常黏膜组织高,差异有统计学意义(1.6±0.7比0.4±0.1,P<0.01);6例结直肠腺瘤组织与对应的正常黏膜中PRL-3mRNA表达量差异无统计学意义(0.6±0.3比0.5±0.2,P>0.05);12例淋巴结转移灶和6例肝转移灶PRL-3基因表达量分别为2.1±0.8和3.3±1.0,显著高于原发癌肿、正常黏膜、阴性淋巴结组织中的表达(P<0.01)。PRL-3基因表达水平与CRCDukes分期、浸润深度、淋巴结和远处转移有关(P<0.05),与性别、肿瘤大小、分化程度无关(P>0.05)。PCR-SSCP分析发现,6例肝转移灶中有1例出现异常泳动条带。结论PRL-3基因在CRC的发生发展和侵袭转移中起重要作用,其作用可能是通过基因表达上调和基因突变共同实现的。     

趋化因子受体CXCR4在结直肠癌组织中的表达及其与临床病理特征的关系

目的观察趋化因子受体CXCR4在结直肠癌组织中的表达及其临床意义,探讨基质衍生因子SDF-1（CXCL12）-CXCR4生物学轴在结直肠癌侵袭转移中的作用。方法免疫组织化学法检测53例结直肠癌组织及27例正常黏膜组织中CXCR4的表达及分布,并分析高表达CXCR4与结直肠癌患者临床病理特征的关系。反转录-聚合酶链反应（RT—PCR）、Western印迹方法检测27例结直肠癌肿瘤组织、正常黏膜及5例肝转移灶内CXCR4 mRNA表达及CXCR4蛋白表达水平。结果53例结直肠癌组织中CXCR4表达阳性率为73．6％,高表达率（表达超过50％细胞）为45．3％;有淋巴结转移组CXCR4高表达率（65．4％）明显高于无淋巴结转移组（25．9％）（P〈0．01）。并与肿瘤血管淋巴管浸润有关。随着临床分期的增加,肿瘤组织中CXCR4高表达率有升高趋势,但差异无统计学意义（P〉0．05）。CXCR4的表达与患者肿瘤部位、大小、浸润深度无关（P〉0．05）。27例新鲜结直肠癌组织中CXCR4mRNA及蛋白表达水平明显高于相应的正常黏膜组织（P〈0．01）;5例肝转移灶组织中CXCR4表达显著高于对应的原发灶（P〈0．01）。结论趋化因子受体CXCR4是一类参与结直肠癌病程进展的重要分子,CXCL12-CXCR4信号通路有望成为结直肠癌治疗的新靶点。     

Significance of tissue laminin P(1) elastase and fibronectin levels in transitional cell carcinoma of the bladder

OBJECTIVE: Elastase is a serine protease which hydrolyses connective tissue components. Laminin and fibronectin also play an important role in progression and invasion of cancer. The purpose of this study is to investigate the relation between tissue elastase, laminin P(1) and fibronectin levels and tumor characteristics, and analyze the potential of these as prognostic factors in transitional cell carcinoma (TCC) of the bladder. METHODS: Thirty-four patients with TCC of the bladder and 11 controls were included in this study. Elastase and fibronectin levels in tissue homogenates were determined using an enzyme immunoassay and laminin P(1) by radioimmunoassay. Mean follow-up was 43 months. RESULTS: The mean elastase level in bladder carcinoma tissue was 120+/-11.42 ng/homogenate protein, while normal tissue level was 12.36+/-2.70 (p<0.01). The carcinoma and normal tissue mean laminin P(1) levels were 7.02+/-0.37 U and 0.65+/-0.10 U/mg homogenate protein, respectively (p<0.01). The mean fibronectin level was 19.97+/-1.45 ng/mg homogenate protein in the carcinoma tissue and 2.16+/-0.40 in normal tissue (p<0.01). There was no correlation between tumor stage, grade, size, multiplicity and elastase, laminin P(1) and fibronectin levels. CONCLUSION: These results provide evidence that tissue elastase, laminin P(1) and fibronectin levels increase in TCC of the human bladder. Further studies including serum and urine levels should be performed in order to analyze their value as tumor markers in a larger group of patients.     

The effect of calcitonin and growth hormone on urinary deoxypyridinoline levels in burned patients

Reduced bone formation and bone loss have been documented in patients following burn injury. Urinary deoxypyridinoline (DPD) is accepted as a marker of collagen breakdown activity. Because calcitonin (CT) diminishes bone resorption and growth hormone (GH) increases bone formation and density in GH-deficient patients, we studied the short-term effects of CT and GH on urinary DPD levels in burned patients. In 30 patients with severe burns, urinary DPD levels were investigated for 3 days following hospitalisation. Then the patients were divided into 3 groups of 10. In the CT group, CT 100U was injected subcutaneously daily for 5 days. In the GH group, GH 0.1mg/kg was injected subcutaneously three times in a week. In the control group, isotonic saline solution 0.1mg/kg was injected subcutaneously three times in a week. In all groups, following the last dose of the agents, urinary DPD levels were investigated for 3 days again. Mean burn size and age were not significantly different between the groups. Urinary DPD level obtained in the early period was 16.5 +/- 3.1nM in the CT group, 10.4 +/- 5.3nM in the GH group and 18.6 +/- 2.7nM in the control group. There were no statistical differences among the groups (P > 0.5, for all). Urinary DPD level obtained in the late period was 4.5 +/- 1.0nM in the CT group, 14.4 +/- 5.9nM in the GH group and 36.6 +/- 2.1nM in the control group. The differences between the CT group and control group, the CT group and GH group and the GH group and control group were statistically significant (P < 0.001, P < 0.01, P < 0.01, respectively). In the comparison of early and late urinary DPD levels, a significant decrease was only obtained in the CT group (P < 0.001, Z:6.5). In the other 2 groups, DPD levels increased in the late period. We concluded that GH is not effective in decreasing urinary DPD levels. On the contrary, CT was found to very effective in decreasing urinary DPD levels. This decrease in urinary DPD levels may be associated with diminished bone loss     

Effect of blood flow occlusion on laser hyperthermia for liver metastases

BACKGROUND: Interstitial laser hyperthermia (ILH) is an in situ ablative technique for the treatment of colorectal liver metastases. A significant factor limiting tumor destruction is hepatic blood flow. Modulation of hepatic blood flow may increase the size of tumor necrosis achieved. Our aim was to investigate the effect of blood flow occlusion on ILH-induced necrosis in both tumor and normal liver tissue. MATERIALS AND METHODS: A model of colorectal liver metastases in male inbred CBA mice was used. ILH was applied to normal liver and tumor tissue using a bare optical quartz fiber from an SYL500 Nd:YAG surgical laser generator, with and without hepatic blood flow occlusion, and the extent of necrosis was studied. Tumor blood flow was assessed by laser Doppler flowmetry and scanning electron microscopy. RESULTS: Hepatic blood flow occlusion resulted in a significant reduction in blood flow in normal liver tissue (37.9% +/- 5.8, P < 0.001) and in the periphery of the tumor (17.5% +/- 7.8, P < 0.001). It did not affect the blood flow in the center of the tumor (13.4% +/- 4.3, P = 0.07). ILH of normal liver tissue, at low power (2 W), with hepatic blood flow occlusion, resulted in a significant increase in the diameter of necrosis. This effect was not seen when higher power (5 W) was used in normal liver. No significant effect was noted within tumor tissue at either power setting. CONCLUSION: The overall effect of hepatic blood flow occlusion in ILH-induced tissue necrosis appears to be negligible in tumor tissue. Its main applicability appears to be at the tumor-host interface, where a decrease in blood flow may lead to higher temperatures and therefore to a greater degree of tumor cell destruction.     

Evaluation of hematuria using the urinary albumin-to-total-protein ratio to differentiate glomerular and nonglomerular bleeding

BACKGROUND: Depending on the etiology and pathophysiology of hematuria, urinary bleeding is classified as glomerular hematuria or nonglomerular hematuria. Nephritis is usually detected by the presence of proteinuria, especially elevated albumin excretion. In this study, we report on the use of the urinary albumin-to-total-protein ratio to accurately differentiate glomerular and nonglomerular bleeding. METHODS: A total of 143 fresh, random urine specimens demonstrating microscopic hematuria (5 or more red blood cells per high-power field) from patients with the source of the hematuria confirmed by histopathology and/or clinical criteria were included in the study. RESULTS: Of the 143 specimens, 104 were from patients diagnosed with glomerular disease and 39 were from patients with nonglomerular disease. Corrected for urine concentration, the mean total-protein-to-creatinine (Cr) and albumin-to-Cr ratios in the glomerular disease group were 1.67 +/- 2.71 g/g Cr and 1.15 +/- 1.77 g/g Cr, respectively (P < 0.001). In the nonglomerular group, the mean total protein-to-Cr and albumin-to-Cr ratios were 0.19 +/- 0.23 g/g Cr and 0.05 +/- 0.06 g/g Cr, respectively (P < 0.001). However, considerable overlap in the ratios among glomerular and nonglomerular disease groups was observed. In contrast, the mean albumin-to-total protein ratios for glomerular and nonglomerular diseases were 0.72 +/- 0.10 and 0.35 +/- 0.17, respectively (P < 0.001). At a cutoff of 0.59, the albumin-to-total-protein ratio demonstrated a sensitivity of 97.1% (101 of 104 cases) in detecting glomerular disease. CONCLUSIONS: The urinary albumin-to-total-protein ratio is potentially a useful index for the differentiation of glomerular and nonglomerular disease in the presence of microscopic hematuria.     

Matrix metalloproteinase 2 and 9 activity in patients with colorectal cancer liver metastasis

BACKGROUND: Matrix metalloproteinases (MMPs) have been reported to play an important role in tumour cell invasion and metastasis. The bioactivity of MMPs in liver metastasis from colorectal cancer was investigated and correlated with clinicopathological variables. METHOD: Thirty-two patients underwent resection of colorectal cancer liver metastases. Latent and active forms of MMP were measured in tissue extracts, by means of quantitative gelatin zymography and a fluorometric activity assay. RESULTS: Broad-spectrum MMP activity, and levels of both active and latent forms of MMP-2 and MMP-9, were higher in tissues containing metastatic tumour than in normal liver tissue. Median metastatic to normal tissue ratios were 15.0 and 17.6 for active and proMMP-2 respectively, and those for active and proMMP-9 were 6.2 and 2.9. The ratios of active to latent enzyme were higher in metastatic tissue than in normal tissue. Lowered MMP-2 activity was associated with large metastatic lesions and increased proMMP-9 levels with preoperative chemotherapy. Both MMP-2 and MMP-9 activity were linked unfavourably to early recurrent disease. CONCLUSION: These data suggest a role for MMPs in colorectal cancer liver metastasis, but indicate different roles for individual MMPs.