微血管密度及增殖细胞核抗原表达与宫颈癌放射敏感性的关系初探

目的探讨增殖细胞核抗原(PCNA)和肿瘤间质微血管密度(MVD)与宫颈癌放射敏感性的关系.方法采用回顾性研究的方法,运用LSAB免疫组织化学染色技术检测80例单纯放射治疗并随访5年以上宫颈癌患者的PCNA指数和MVD值.结果 80例宫颈癌患者均有不同程度的PCNA表达,PCNA指数分布在0.243～0.913之间,平均为0.522;微血管计数为(51.8±11.5)条,实际范围21～72条.结论 PCNA的表达和MVD值与宫颈癌的放射敏感性有关,为寻找能预测宫颈癌放射敏感性的肿瘤标志物,指导临床治疗方案的选择提供了初步的实验依据.     

新辅助化疗对宫颈癌微血管和微淋巴管的影响及疗效分析

目的:探讨新辅助化疗对局部晚期宫颈癌微血管和微淋巴管的影响及疗效。方法:选择70例ⅠB2～ⅡB期行新辅助化疗的宫颈癌组织,采用免疫组织化学方法检测其中微血管密度(microvessel density,MVD)和微淋巴管密度(lymphatic microvessel density,LMVD),并分析化疗疗效与临床病理因素的关系。结果:宫颈癌组织中MVD和LMVD水平均高于正常组织,差异均有统计学意义(P<0.001,P=0.028);MVD水平与肿瘤浸润深度有关(P=0.039),LMVD水平与鳞状细胞癌抗原(squamous cell carcinoma antigen,SCCAg)水平(P=0.017)和淋巴结转移(P=0.049)有关。化疗后MVD和LMVD均比化疗前降低,前者差异有统计学意义(P=0.019),而后者差异尚无统计学意义(P=0.176)。新辅助化疗总有效率为70.0%(49/70),总完全缓解率为21.4%(15/70);多因素分析显示,宫颈癌浸润深度是影响化疗疗效的独立因素(P=0.018,OR=0.169)。结论:新辅助化疗可有效降低局部晚期宫颈癌的MVD水平,缩小肿瘤体积。     

胸腺嘧啶磷酸化酶在结直肠癌组织中的表达

目的研究胸腺嘧啶磷酸化酶(thymidine phosphorylase,TP)在结直肠癌组织中的表达,为临床用药提供指导.方法运用实时荧光定量RT-PCR和ELISA方法分别从mRNA和蛋白质水平对TP在结直肠癌组织及相邻正常组织中的表达进行检测.结果无论是在mRNA还是蛋白质水平,肿瘤组织中的TP表达高于其相邻正常组织,肿瘤组织中的TP蛋白水平为7.82±4.31 ng/μgTPO,mRNA水平为1.66±0.766;正常组织中TP蛋白水平为5.03±2.48 ng/μgTPO,mRNA水平为1.05±0.60;并且ELISA结果与实时RT-PCR结果具有一定的相关性(γ=0.81).结论胸腺嘧啶磷酸化酶在结直肠肿瘤中的表达对于临床使用5-FU类药物具有指导意义.     

胸苷磷酸化酶在胃癌中的表达及临床意义

目的：探讨胸苷磷酸化酶（ＴＰ）在胃癌中的表达及临床意义。方法：采用免疫组织化学法检测２０４例胃癌患者肿瘤组织中ＴＰ的表达,并探讨与临床病理因素、术后辅助化疗和预后之间的关系。结果：胃癌组织ＴＰ表达阳性率为４７.１％。ＴＰ阳性表达患者中位生存时间为３０.９４个月,ＴＰ阴性表达患者为３４.４３个月,两者比较有显著的统计学差异（Ｐ＝０.００２）。ＴＰ阳性表达与患者年龄、发病部位、ＴＮＭ分期、浸润深度、淋巴结转移等病理因素以及与以氟尿嘧啶为主的辅助化疗的疗效无明显相关性。结论：ＴＰ表达与胃癌患者长期生存有一定的相关性,有可能成为预测胃癌预后的一个参考指标。     

Keratin expression in normal uterine cervical epithelium and carcinomas of cervical origin

The immunohistochemical expression of keratins 7, 8, 10, 13, 14, 17, 18 and 19 was examined in formalin-fixed paraffin-embedded tissues of normal uterine cervical epithelium and carcinomas of cervical origin (4 squamous cell carcinoma in situ, 17 squamous cell carcinoma, 9 adenocarcinoma, and 1 adenoid basal carcinoma). A panel of 8 monoclonal antibodies capable of recognizing 8 individual keratin subtypes was employed using microwave oven heating and a labeled streptavidin biotin method. Ectocervical squamous epithelium expressed keratins 14 and 19 in the basal cell layer, and keratins 10 and 13 in the suprabasal cell layer. Endocervical columnar cells were found to express keratins 7, 8, 18 and 19, whereas the reserve cells expressed keratins 7, 8, 14, 17, 18 and 19. Most of the squamous cell carcinomas, both keratinizing and non-keratinizing, as well as the carcinoma in situ revealed a keratin phenotype detected in normal ectocervical squamous cells (keratins 10, 13, 14 and 19) and endocervical subcolumnar reserve cells (keratins 7, 17 and 18). The adenocarcinomas, both endocervical and endometrial type, were positive for keratins 7, 8, 17, 18 and 19. The adenoid basal carcinoma expressed all the keratins examined including the expression of reserve cell keratin. Reserve cell keratins were found mostly in squamous cell carcinomas, adenocarcinomas and adenoid basal carcinoma of cervical origin. Therefore, the keratin expression pattern indicates the origin of a variety of carcinomas of the uterine cervix from a common progenitor, endocervical reserve cells.     

Thymidine phosphorylase expression correlates with tumor differentiation and bcl-2 in invasive breast cancer

BACKGROUND: Angiogenesis plays an important role in the growth and metastasis of solid tumors. Several angiogenic factors have been identified, and thymidine phosphorylase (TP) is thought to be one such factor. To date, little information is available on the relationship between TP and other clinicopathological variables. METHODS: Formalin-fixed, paraffin-embedded materials from 116 primary breast carcinomas were used. The expression of TP, estrogen receptor, Bcl-2, Bax, p53, c-erbB-2 and MIB-1 was examined by immunohistochemical methods. RESULTS: Nuclear and/or cytoplasmic TP expression was observed in the neoplastic cells, and accentuation of TP was often present at the infiltrating tumor edge and intraductal spread region. Tumor cell TP expression was significantly inversely correlated with histological grade (p< 0.05) and positively correlated with Bcl-2 expression, but no association with other tumor variables was found. CONCLUSIONS: TP is associated with Bcl-2 expression and tumor differentiation in breast cancer. TP may be a new prognostic parameter for breast cancer.     

Thymidine phosphorylase expression is preserved after radiotherapy in patients with cervical squamous cell carcinoma

Purpose The aim of this study was to investigate the changes in two of the enzymes involved in fluorouracil metabolism, thymidine phosphorylase (TP) and dihydropyrimidine dehydrogenase (DPD), in uterine cervical squamous cell cancer tissue after radiotherapy.Subjects and methods Cervical tissue from 27 patients diagnosed with stage IIIB or IV uterine cervical squamous cell cancer was compared with normal cervical tissue from 33 patients with benign gynecologic diseases. Expression of TP and DPD in the cervical tissues was measured using enzyme-linked immunosorbent assays. TP and DPD expression before and after irradiation with 10 and 20 Gy was measured in 9 of the 27 patients with cervical cancer.Results Before irradiation, DPD expression in cancer tissue did not differ from that in normal tissue. TP expression and the TP/DPD ratio were significantly higher in cancer tissue than in normal tissue (P<0.00001). TP and DPD expression and the TP/DPD ratio were not significantly changed by irradiation with 10 and 20 Gy. TP expression and the TP/DPD ratio after irradiation with 10 and 20 Gy were significantly higher than in normal tissue.Conclusion The increased TP expression and the elevated TP/DPD ratio following irradiation with up to 20 Gy may offer an increased clinical advantage to chemoradiotherapy with capecitabine or doxyfluridine over radiotherapy alone.     

Continuous representation of tumor microvessel density and detection of angiogenic hotspots in histological whole-slide images

Blood vessels in solid tumors are not randomly distributed, but are clustered in angiogenic hotspots. Tumor microvessel density (MVD) within these hotspots correlates with patient survival and is widely used both in diagnostic routine and in clinical trials. Still, these hotspots are usually subjectively defined. There is no unbiased, continuous and explicit representation of tumor vessel distribution in histological whole slide images. This shortcoming distorts angiogenesis measurements and may account for ambiguous results in the literature.In the present study, we describe and evaluate a new method that eliminates this bias and makes angiogenesis quantification more objective and more efficient. Our approach involves automatic slide scanning, automatic image analysis and spatial statistical analysis. By comparing a continuous MVD function of the actual sample to random point patterns, we introduce an objective criterion for hotspot detection: An angiogenic hotspot is defined as a clustering of blood vessels that is very unlikely to occur randomly. We evaluate the proposed method in N=11 images of human colorectal carcinoma samples and compare the results to a blinded human observer. For the first time, we demonstrate the existence of statistically significant hotspots in tumor images and provide a tool to accurately detect these hotspots.     

Thymidine phosphorylase expression in oral squamous cell carcinoma

Thymidine phosphorylase (TP), as an enzyme involved in DNA synthesis, catalyzes the reversible conversion of thymidine to thymine. It is also identical to the angiogenic factor, platelet-derived endothelial cell growth factor. We examined TP expression using immunohistochemistry in 66 archival samples obtained from the patients with primary oral squamous cell carcinoma (SCC) and investigated its relation to tumor vascularity, cell proliferation, apoptosis, clinicopathological features and survival. TP expression was identified in cytonucleus and/or cytoplasm in carcinomas, but was not identified in histologically normal epithelia distant to tumor in most cases. No significant difference of microvessel density (MVD) was found between the carcinomas with high TP expression (H-TP) and low TP expression (L-TP). The percentages of proliferative cells marked by Ki-67 staining in H-TP carcinomas was significantly higher than that in L-TP carcinomas (P=0.0222). The apoptotic indice (AI) in H-TP carcinomas tended to be lower than that in L-TP carcinomas (P=0.0723). Moreover, the level of TP expression was significantly correlated the pattern of tumor invasion (P=0.0146) and marginally correlated with lymph nodal metastasis (P=0.0804). Our results suggested that TP enzyme may play a role in promotion of tumor growth in oral SCC, and that its expression can be indicative of tumor aggressiveness in this tumor type.     

卵巢浆液性癌中COX-2表达与微血管密度的相关性

目的:探讨卵巢浆液性癌组织中环氧化酶2（COX-2）的表达与癌组织中微血管密度（microvessel density,MVD）的相关性。方法:应用免疫组织化学染色SP法检测94例卵巢浆液性癌组织中COX-2和CD34的表达,分析癌组织中COX-2的表达与癌组织中MVD的相关性。结果:卵巢浆液性癌组织中COX-2阳性表达56例,阴性表达38例。卵巢浆液性癌94 例MVD的中位数为34.33,以34.33为分界线,将其分为两组,高MVD组（44例）和低MVD组（50例）。高MVD组,中位生存时间为16个月,3年生存率为54.55%,5年生存率为27.27%,低MVD组中位生存时间为38个月,3年生存率为78.00%,5年生存率为62.00%。两组间比较,预后差异有统计学意义(P＜0.05);COX-2的表达与MVD值呈正相关性（P＜0.05）。结论:卵巢浆液性癌中COX-2的表达与新生血管相关,可促进肿瘤的发生、发展。     

Thymidine phosphorylase activity required for tumor angiogenesis and growth

Thymidine phosphorylase (TP) is identical to platelet-derived endothelial cell growth factor (PD-ECGF), and has angiogenic activity. We examined the involvement of TP activity in tumor growth and angiogenesis. KB cells were transfected with wild-type or mutant (L148R) PD-ECGF cDNA, and two sublines with high TP activity, KB/wt4 and KB/wt6, and one subline with no TP activity, KB/L148R, were cloned, respectively. The doubling times of these subclones in vitro were similar to that of KB cells. However, the growth of KB/wt4 and KB/wt6 cells was significantly faster when xenografted into nude mice than that of control cells with no TP activity. The tumors with high TP activity (KB/wt4 and KB/wt6) had significantly more microvessels than those with no TP activity (KB/-, KB/CV and KB/L148R) (P<0.01). These results, taken together with previous reports, suggest that the TP enzyme activity itself is involved in angiogenesis and growth of the KB tumors.     

Correlation between microvessel density and tumor cell proliferation with clinical factors in breast carcinomas.

Fifty-nine methacarn-fixed, paraffin-embedded human breast carcinomas were immunostained by QB-END/10 (an antibody to CD34 antigen) to observe microvessels and by PC10 (an antibody to proliferating cell nuclear antigen; PCNA) to determine tumor cell proliferation; 9 normal human breast tissue specimens were also immunostained by QB-END/10. The number of microvessels in the periphery of the breast carcinoma was significantly greater than both that in the center of the breast carcinoma and that in the normal breast. There was also a significant relationship between the number of microvessels in the periphery of breast carcinomas and the histological tumor size and lymph node status. However, there was no significant relationship between the tumor cell proliferation activity (PCNA positive cell ratio) and any clinical or histopathological variables. The number of microvessels and the tumor cell proliferation activity showed a weak negative correlation.     

Expression of thymidine phosphorylase correlates with microvessel density in prostate cancer

Angiogenesis in the growth and development of prostate cancer was the focus of this study. Various angiogenic factors and their clinicopathologic correlations with the progression of prostate cancer have been examined. Thymidine phosphorylase is identical to platelet-derived endothelial cell growth factor (TP/PD-ECGF) and has angiogenic activity. We investigated the expression of TP/PD-ECGF in prostate cancer and its association with angiogenesis or clinicopathologic findings in 81 cases with prostate cancer. Western blot analysis using a specific monoclonal antibody 654-1 revealed the existence of a 55 kDa TP/PD-ECGF protein in human prostate cancer tissue. Cancer tissue showed low-positive immunostaining in 32 cases (39.5%) and high positivity in 49 cases (60.5%). This protein expression indicated a statistically significant association with microvessel density (low vs. high TP/PD-ECGF expression group: mean +/- SD, 37.3+/-27.0 vs. 53.1+/-28.0 microvessels in three fields, p<0.05). No correlation was found between the expression of TP/PD-ECGF and nuclear grade, glandular differentiation, clinical stage or overall survival rate. TP/PD-ECGF may play an important role in tumor angiogenesis in prostate cancer tissues. Although the expression of TP/PD-ECGF was not correlated with clinical outcome in patients with prostate cancer, there remains the possibility that TP/PD-ECGF may support or modify the tumor growth through angiogenesis in cooperation with other factors.     

Thymidine phosphorylase expression is associated with both increase of intratumoral microvessels and decrease of apoptosis in human colorectal carcinomas.

Thymidine phosphorylase (dThdPase)/platelet-derived endothelial cell growth factor is expressed at higher levels in a variety of human carcinomas than it is in adjacent normal tissue. The higher expression is associated with an increase of intratumoral microvessel density (IMVD) and an unfavorable patient prognosis. We examined the role of dThdPase in apoptosis, cell proliferation, IMVD, and p53 expression in human colorectal carcinomas. dThdPase expression was noted in 13 of 36 (36.1%) Dukes' A and B carcinomas and in 13 of 28 (46.3%) Dukes' C and D carcinomas. At least 10 areas consisting of carcinoma cells with diffuse dThdPase expression from the 26 dThdPase-positive tumors (category I) and 10 areas without dThdPase expression from the 38 negative tumors (category II) were selected from each case. For stage A and B tumors, the mean IMVDs were 64.8 +/- 33.7 in category I and 33.2 +/- 12.6 in category II tumors, whereas for stage C and D tumors, the mean IMVDs were 77.6 +/- 27.2 in the category I and 34.7 +/- 14.0 in the category II tumors. The mean IMVD was significantly higher in category I than in category II tumors (P < 0.01). The mean apoptotic indices (AIs; percentage of apoptotic cells) were 2.7 +/- 1.7 in the category I and 5.4 +/- 2.2 in the category II carcinomas of stages A and B and 1.4 +/- 0.5 in category I and 5.3 +/- 2.3 in category II carcinomas of stages C and D, and the value of the mean AI was significantly lower in category I than in category II (P < 0.01), regardless of the Dukes' stage. AI and IMVD showed a significant inverse correlation (P < 0.001). There was no significant difference in the frequency of p53 expression between the two categories. These results indicated that dThdPase expression provides an advantage for tumor growth of human colonic carcinomas not only by increasing the intratumoral microvessels but also by attenuation of apoptosis, which might occur via a p53 gene-independent pathway.     

pS2 expression in primary breast carcinomas: Relationship to clinical and histological features and survival

Summary pS2 protein expression has been reported to have prognostic significance in human breast carcinomas and to correlate with estrogen receptor positivity, although these findings have not been confirmed by all investigators. pS2 positivity was compared to various clinical and histologic parameters in a retrospective study of 290 patients (median follow-up 7.2 years) and significantly correlated with tumor grade and estrogen receptor content (p=0.001 and p=0.0007, respectively). Significant associations between pS2 positivity and lymph node metastases, T stage, histologic tumor type, and patient age were not observed. Univariate and multivariate analyses (controlling for estrogen receptor content, T and N stage) of the patient population at large showed that pS2 positivity was not predictive of disease-free or overall survival. Univariate analysis of lymph node negative patients demonstrated that both pS2 and estrogen receptor positivity were significantly associated with a better outcome. Multivariate analysis of these patients, however, showed that only estrogen receptor data had independent prognostic significance. This study suggests that immunohistochemical analysis for pS2 protein expression will not contribute additional prognostic information if the estrogen receptor content is known.     

Macrophage infiltration-associated thymidine phosphorylase expression correlates with increased microvessel density and poor prognosis in astrocytic tumors.

PURPOSE AND EXPERIMENTAL DESIGN: To clarify the significance of thymidine phosphorylase (TP)/platelet-derived endothelial cell growth factor/gliostatin in human glioma, we examined TP expression immunohistochemically in a series of 50 astrocytic tumors and correlated its expression with tumor angiogenesis and apoptosis, as well as prognosis. RESULTS: The majority of TP-positive cells were of macrophage origin, which was confirmed by immunostaining TP and CD68 on mirror sections. TP expression was significantly associated with glioma malignancy grading, intratumoral microvessel density, and VEGF expression but showed no relationship with apoptotic index or P53 expression. Regardless of glioma grading, patients with TP-positive tumors had a significantly shorter mean survival time than those with TP-negative tumors. CONCLUSIONS: These findings suggest that TP might play a crucial role in angiogenesis during glioma development, and immunodetection of TP is useful for clinical prediction. Further studies are necessary to better elucidate the role of TP in glioma, which may provide insights into adequate TP-targeted therapy.     

Thymidine phosphorylase and dihydropyrimidine dehydrogenase protein expression in colorectal cancer.

It is essential for actively proliferating cells to increase their rate of DNA synthesis to progress through the cell cycle. This is reflected in the increased uracil usage that is a common feature in solid tumours. Thymidine phosphorylase (TP) anabolises formation of pyrimidine nucleosides available for DNA synthesis, whereas dihydropyrimidine dehydrogenase (DPD) catabolises the degradation of pyrimidine bases, thereby reducing levels of uracil and thymine available for DNA synthesis. In addition, tissue levels of TP or DPD have been associated with the clinical efficacy of pyrimidine anti-metabolites commonly used in the treatment of colorectal cancer. There is little information, however, on the relative expression or degree of co-ordinated regulation of either protein in primary or metastatic colorectal cancer. DPD and TP protein levels were measured in 15 primary colorectal carcinomas, 10 colorectal liver metastases and 25 adjacent uninvolved tissues. DPD was reduced in 67% (10/15) of colorectal tumours (mean tumour/normal = 0.52) and in all liver metastases (mean tumour/normal = 0.41) compared with the corresponding normal tissue. In contrast, TP was increased in 80% (12/15) of colorectal tumours (mean tumour/normal = 18.91) and in all metastases (mean tumour/normal = 3.70). TP and DPD protein expression were highly variable in uninvolved and tumour tissues. The ratio of TP:DPD was higher in 87% of colorectal tumours and in all liver metastases compared with the adjacent uninvolved tissues. This suggests the presence of co-ordinated regulation of these pyrimidine metabolic enzymes and offers a strategy for optimising the use of pyrimidine-based chemotherapy.     

Thymidine phosphorylase expression and effect of doxifluridine: a phase II study.

Doxifluridine (5'-DFUR), an active intermediate metabolite of capecitabine, is converted to 5-fluorouracil by thymidine phosphorylase (TP). We used immunohistochemical staining to investigate the relation between TP expression and 5'-DFUR effects in 40 patients with advanced/recurrent lung metastases from colorectal cancer. Cox regression analysis suggested that TP-positive cancer cells (risk ratio 3.72), were independent factors in survival whereas factors in progression-free survival were TP-positive cancer cells (2.93), and TP-positive stromal cells (0.24). It is suggested that TP expression in cancer cells and in stromal cells are opposite prognostic factors in patients treated with 5'-DFUR.     

Thymidine phosphorylase expression in hepatic metastasis from colorectal carcinoma]

Thymidine phosphorylase (dThdPase) is identical to platelet-derived endothelial cell growth factor (PD-ECGF), and has angiogenic activity. It has been reported that a higher level of dThdPase activity is associated with an unfavorable patient prognosis. The activity of dThdPase was investigated in 15 patients who underwent hepatectomy for hepatic metastasis from colorectal carcinoma, and an assessment was made as to whether its expression was correlated with the prognosis. The dThdPase activity was significantly higher in the patients with extrahepatic metastasis after hepatectomy (n = 5) than in those without extrahepatic metastasis (n = 10) (168.2 +/- 50.8 vs 95.1 +/- 35.2 Unit/mg prot., p < 0.05). This dThdPase activity could be a significant prognostic indicator, and was useful in predicting the extrahepatic recurrence after hepatectomy for hepatic metastasis from colorectal carcinoma.     

Clinical relevance of microvessel density in laryngeal squamous cell carcinomas

The clinical implications of microvessel density (MVD) in head and neck tumors have not been fully elucidated. We investigated the clinicopathologic correlates and prognostic relevance of MVD in a series of 122 consecutive patients with surgically treated laryngeal squamous cell carcinoma followed-up for a mean of 79 months. MVD was evaluated after CD34 immunostaining in 3 250x microscopic fields representative of the "hot spot" area, and expressed as the mean value of the vessel counts per millimeter squared. The overall median value of the intratumoral vessel count was 69.5/mm(2). In the 20 cases we analyzed, MVD increased significantly from normal to dysplastic mucosa and infiltrating carcinoma (p = 0.0001). Nineteen carcinomas (15.6%) had MVD values that were equal to or lower than the highest MVD value (52.7/mm(2)) observed in normal mucosa samples (in which the median MVD count was 34.5/mm(2), range 16.6-52.7/mm(2), mean 35.1 +/- 11.5/mm(2)) and were therefore considered poorly vascularized. Periodic acid-Schiff (PAS) staining revealed intratumoral PAS-positive connective tissue septa in 13 cases (10.7%). The patients with poorly vascularized tumors showed a tendency toward a better prognosis, but the anatomical site, tumor extension and clinical stage were the only variables significantly associated with disease-free and overall survival.