Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina / Propiedades inhibidoras Enzimáticas y Anticancerígenas de la Moringa peregrina

Background: Moringa peregrina is widely used in the traditional medicine of the Arabian Peninsula to treat various ailments, because it has many pharmacologically active components with several therapeutic effects. Objective: This study aimed to investigate the inhibitory effect of Moringaperegrina seed ethanolic extract (MPSE) against key enzymes involved in human pathologies, such as angiogenesis (thymidine phosphorylase), diabetes (α-glucosidase), and idiopathic intracranial hypertension (carbonic anhydrase). In addition, the anticancer properties were tested against the SH-SY5Y (human neuroblastoma). Results: MPSE extract significantly inhibited α-glucosidase, thymidine phosphorylase, and carbonic anhydrase with half-maximal inhibitory concentrations (IC50) values of 303.1 ± 1.3, 471.30 ± 0.3, and 271.30 ± 5.1 µg/mL, respectively. Furthermore, the antiproliferative effect of the MPSE was observed on the SH-SY5Y cancer cell line with IC50 values of 55.1 µg/mL. Conclusions: MPSE has interesting inhibitory capacities against key enzymes and human neuroblastoma cancer cell line.

Antecedentes: La Moringa peregrina se utiliza ampliamente en la medicina tradicional de la Península Arábiga para tratar diversas dolencias, ya que posee numerosos componentes farmacológicamente activos con varios efectos terapéuticos. Objetivo: Este estudio tenía como objetivo investigar el efecto inhibidor del extracto etanólico de semillas de Moringaperegrina (MPSE) frente a enzimas clave implicadas en patologías humanas, como la angiogénesis (timidina fosforilasa), la diabetes (α-glucosidasa) y la hipertensión intracraneal idiopática (anhidrasa carbónica). Además, se comprobaron las propiedades anticancerígenas frente al SH-SY5Y (neuroblastoma humano). Resultados: El extracto de MPSE inhibió significativamente la α-glucosidasa, la timidina fosforilasa y la anhidrasa carbónica con concentraciones inhibitorias semimáximas (IC50) de 303,1 ± 1,3, 471,30 ± 0,3 y 271,30 ± 5,1 µg/mL, respectivamente. Además, se observó el efecto antiproliferativo del MPSE en la línea celular del cáncer SH-SY5Y con valores de IC50 de 55,1 µg/mL. Conclusiones: MPSE posee interesantes capacidades inhibitorias frente a enzimas clave y línea celular de neuroblastoma canceroso humano.

Choice of Capecitabine or S1 in Combination with Oxaliplatin based on Thymidine Phosphorylase and Dihydropyrimidine Dehydrogenase Expression Status in Patients with Advanced Gastric Cancer

PURPOSE: To study the efficacy of capecitabine or S-1 plus oxaliplatin (CAPOX or SOX) for treating thymidine phosphorylase (TP)- or dihydropyrimidine dehydrogenase (DPD)-positive advanced gastric cancer.MATERIALS AND METHODS: Eighty-six patients with stage IIIC to IV gastric cancer were assessed for TP and DPD expression by immunohistochemistry. The association between CAPOX or SOX efficacy and TP/DPD expression was retrospectively analyzed.RESULTS: There were no significant differences in the objective remission rate (ORR, 52.27% vs. 47.62%; P>0.05), disease control rate (72.73% vs. 73.81%, P>0.05), progression-free survival (hazard ratio [HR], 1.119; 95% confidence interval [CI], 0.739–1.741; P=0.586), and overall survival (OS; HR, 0.855; 95% CI, 0.481–1.511; P=0.588) between CAPOX and SOX. A higher number of stage IV patients showed TP positivity, while DPD-positive patients predominantly showed intestinal type of gastric cancer. In TP-positive patients, the ORRs associated with CAPOX and SOX treatments were 57.14% and 38.10%, respectively; OS was better with CAPOX than with SOX (HR, 0.447; 95% CI, 0.179–0.978; P=0.046). Among DPD-positive patients, the SOX treatment-associated ORR (60.87%) was significantly higher than the CAPOX treatment-associated ORR (43.48%). Furthermore, SOX treatment resulted in better OS than did CAPOX treatment (HR, 2.020; 95% CI, 1.019–4.837; P=0.049).CONCLUSIONS: No significant difference in clinical efficacy was found between CAPOX and SOX. TP-positive patients might respond better to CAPOX while DPD-positive patients may respond better to SOX. Our findings might serve as a guide for personalized chemotherapy for gastric cancer.

Influence of thymidine phosphorylase polymorphisms on the efficacy of capecitabine-based adjuvant chemotherapy in colorectal cancer patients / 中国肿瘤临床

Objective: To inrestigate the association between thymidine phosphorylase (TYMP) polymorphisms and efficacy of postop-erative capecitabine-based adjuvant chemotherapy in colorectal cancer (CRC) patients. Methods: Two hundred and thirty-five patients with colorectal cancer who received surgical treatment and adjuvant chemotherapy between January 2010 and December 2016 from People's Hospital of Zhengzhou, were included in this study. Peripheral blood and postoperative tissue specimens of the CRC patients were collected for genotyping polymorphisms and measuring TYMP mRNA expression, respectively. The correlation between the poly-morphisms and efficacy of postoperative chemotherapy in CRC patients was analyzed. Results: The prevalence of 5633C>T in TYMP gene among the CRC patients was as follows: CC genotype, 149 cases (63.40%); CT genotype, 73 cases (31.06%); and TT genotype, 13 cases (5.54%); the minor allele frequency of 5633C>T was 0.21. Survival analysis of the patients revealed that the median overall sur-vival (OS) of patients with the CT/TT genotype and those with the CC genotype was 5.9 and 4.5 years, respectively; the result was sta-tistically significant (P=0.009). Following adjustment in multivariate Cox regression analysis, the CT/TT genotype was found to be an in-dependent favorable factor for OS (HR=0.67, P=0.015). Additionally, of the 87 postoperative tissue specimens, results show that the levels of TYMP mRNA in cancer tissues of patients with the CT/TT genotype were significantly higher than those with the CC genotype (P=0.019). Conclusions: TYMP mRNA expression may be influenced by the 5633C>T polymorphism, making CRC patients benefit from capecitabine treatment.

Transfected thymidine phosphorylase cDNA with lentiviral vector and INF-α2a enhance the anticancer effects of 5'-deoxy-5-fluorouridine on colorectal cancer cell line LOVO / 中华普通外科杂志

Objective Thymidine phosphorylase (TP) cDNA was transfected into colorectal cancer cell lines LOVO with the lentiviral vector,the anticancer effeciency of 5'-deoxy-5-fluorouridine (5'-DFUR)and 5-fluorouracil (5-FU) on LOVO cells were evaluated.Methods TP cDNA were transfected into LOVO cell line with the lentiviral vector pLenti6.3_MCS_IRES2-EGFP,and the transfection efficiency was analyzed by flow cytometer and immunohistochemistry.Cells were divided into six groups:LOVO,LOVO-TP,LOVO-control,LOVO + INF-α2a,LOVO-TP + INF-α2a,LOVO-control + INF-α2a.TP protein expression and the relative quantitative analysis for TP mRNA in transfections cells were detected by Western blot and RT-PCR respectively.Volumes of converted 5-FU from either in the medium containing different concentration of 5'-DFUR,in which all cells were cultured,or in cells lysate,were detected by high performance liquid chromatography (HPLC).Results The transfection efficiency of TP cDNA in LOVO cells was 95％.The Mean gray value of TP protein expression in LOVO-TP and LOVO-TP + INF-α2a were 198.15 folds and 243.22 folds higher than LOVO cell,respectively (P ＜ 0.01).The RQ values of TP mRNA expression in LOVO-TP and LOVO-TP + INF-α2a were also 18.56 folds and 59.61 folds higher than wild LOVO cell,respectively (P ＜ 0.01).The IC50 values of 5'-DFUR on LOVO-TP and LOVO-TP + INF-α2a were 1 660 μ mol/L and 813 μ mol/L,respectively,significantly lower than 4 462.59 μ mol/L in wild LOVO (P ＜ O.01).The 5-FU volumes detected from media contained series concentration of 5'-DFUR for culturing LOVO-TP and LOVO-TP + INF-α2a were 2.0-5.3 folds,and 2.9-10.4 folds more than wild LOVO,respectively (P ＜ 0.01).Conclusions Transfected TP cDNA into colorectal cancer cell line LOVO with lentiviral vector increases the expression of TP mRNA and TP protein and the amount of 5-FU converted from 5'-DFUR,enhancing its anticancer effect.

Advances in the relationship between thymidine phosphorylase ex-pression and colorectal carcinoma / 中国肿瘤临床

10.3969/j.issn.1000-8179.2013.12.014

Correlations between serum levels of TS, TP and clinical outcome in Chinese patients with advanced & nbsp;gastric cancer receiving fluorouracil-based chemotherapy / 中国癌症杂志

10.3969/j.issn.1007-3969.2013.04.009

Antitumor activity of 5' -deoxy-fluorouridine on colon cancer experimental model in BALB/C mice / 国际外科学杂志

ObjectiveTo evaluate the anticancer activity of 5'-dexoxy-fluorouridine on colon cancer experimental models in BALB/C mice,compared with 5'-fluorouracil,an anticancer agent widely used in clinic,meanwhile,examined the conversion of 5'-Dexoxy-fluorouridine to 5' -fluorouracil in cancer tissues and serum of mouse models.MethodsThe xenografts of mouse colon cancer cell line CT 26 were transplantated to cecum in 60 male BALB/C mice.Three days lated,these mice were divided into 3 groups and intro- peritoneally injected:( 1 ) 5' - dexoxy- fluorouridine 0.1 mg/g,(2) 5' - Fluorouracil 0.02 mg/g,(3)0.9％ sodium chloride 0.4 mL (as a control),respectively.Two and three weeks later,6 mice were sacririced in every group respectively to measure the weight of tumors and bodies,to examine the Hb,RBC,WBC,PLT,AST,ALT,UREA,and CREA in blood.The rest 8 mice in each group were fed generally,and the survival time from operation to natural death was recorded.In addition,14 mice with xenografts of CT 26 about 2 weeks,were divided into 2 groups averagely,5' -dexoxy-fluorouridine 0.1 mg/g and 5' -fluorouracil 0.02 mg/g were intro-peritoneally injected respectively.Fifteen min later,the converted 5' -fluorouracil was detected from the blood and tumor tissues in sacrificed mice.ResultsThe lest tumor average weight was found in the mice injected 5 '-dexoxy-fluorouridine,being (0.07 ± 0.12) g and (0.24g ±0.29) g for the mice sacrificed at 2 and 3 weeks later,respectively.The average survival time for rest mice was ( 32.6 ± 8.9) d.The average tumor weight in 5' - fluorouracil group was (0.74 ± 0.43 ) g and ( 1.13 ±0.75) g at 2 and 3 weeks later,and the average survival time for the rest was (22.8 ±5.9)d,respectively.The average tumor weight in the control group was (0.70 ±0.47) g and ( 1.93 ±0.83) g at 2 and 3 weeks,and the average survival time for the rest was ( 17.5 ± 2.8 ) d.Either the average tumor weight or average survival time in the mice of 5 ' -dexoxy-fluorouridine group was significantly differen from either 5' -fluorouracil group or control (P ＜ 0.05 ).However,there was no significant difference for the numbers of WBC,PLC,Hb,and some function examination of liver and kidney among 3 group mice,besides the loss of weights in 5'-fluorouracil group mice after operation and medicine therapy which was significantly obvious than that in 5' -deoxy-fluorouridine and control groups ( P ＜ 0.05 ).In addition,( 54.71 ± 12.82) μg/g 5' -fluorouracil was detected in xenografts of mice injected 5' -dexoxy-fluorouridine 15 min later,which was the 6.20 folds of 5' -fluorouracil detected in serum from sthe ame group,P ＜0.05.However,( 133.35 ±20.69) μg/m 5'-fluorouracil were detected in serum of mice after 5' -fluorouracil were injected 15 min later,which was the 1.55 folds of 5' -fluorouracil detected in the xenografts from same group ( P ＜ 0.05 ).ConclusionsIn colon cancer tissues of mouse experimental models,5' - dexoxy- fluorouridine could be converted effectively to 5'-fluorouracil,an obvious high concentration being detected in serum of mice than in cancer tissues.The anticancer effect of 5'-dexoxy-fluorouridine on mouse colon cancer models was more effective than 5'-fluorouracil,resulting in a longer survival duration,less side effect and no significant injury on liver and kidney functions.However,the mechanism of 5' -dexoxy-fluorouridine converted to 5' -fluorouracil in cancer tissue is needed further investigation.

Relationship between the Expression of Thymidylate Synthase,Thymidine Phosphorylase and Dihydropyrimidine Dehydrogenase and Survival in Epithelial Ovarian Cancer / 华中科技大学学报（医学）（英德文版）

The mRNA and protein expression of thymidylate synthase(TS),thymidine phosphorylase(TP)and dihydropyrimidine dehydrogenase(DPD)and their relationship with prognosis were investigated.Real-time quantitative RT-PCR(Taqman)was used to detect the mRNA expression of TS,TP and DPD in formalin-fixed and paraffin-embedded 106 samples of epithelial ovarian cancer and 29 normal ovaries.A TATA box-binding protein(TBP)was used as an endogenous reference gene.A relationship between TS,TE DPD expression and clinicopathologic features was investigated.The protein location and expression of TS,TP and DPD was examined in the same patients by an avidin-biotin-peroxidase immunohistochemistry.TS and TP mRNA expression levels were significantly higher in tumor group than in normal controls,with the average value of TS and TP mRNA being 6.14±0.62 and 0.59±0.06 in tumor tissue,and 0.71±0.14 and 0.16±0.04 in normal tissue,respectively.DPD mRNA expression levels were significantly lower in tumor group(0.11±0.02)than in normal controls(0.38±0.05).There was statistically significant difference in TS and TP mRNA expression levels among different pathological grades and clinical stages(P<0.05),but histological subtype was not significantly associated with TS and TP mRNA expression.DPD gene expression was not significantly associated with any clinicopathological parameters.Immunohistochemistry revealed that TP protein was mainly distributed in nucleus,and TS and DPD mainly in cytoplasm.The protein expression intensity of TS,TP and DPD was coincided with the mRNA expression levels.It was concluded that TS,TP mRNA and protein expression levels were significantly higher in epithelial ovarian cancer,and DPD mRNA and protein expression levels were significantly lower.The expression levels of TS and DPD were related to the patients' prognosis and survival.Combined gene expression levels of TS,TP and DPD represent a new variable to predict the clinical outcome in ovarian cancer.The association of TS,TP and DPD expression levels with survival suggests an importance of these genes for tumor occurrence and progression.

Expression of thymidine phosphorylase in human metastatic liver cancer tissues from colorectal cancer / 国际外科学杂志

Objective To detect the tbymidine pbospborylnse (TP) expression in metastatic liver cancer tissues from human colorectal cancer by immunohistochemistry, and analyze the correlation between TP ex-pression and the tumor-associated macrophages (TAM), and the prognosis of patients. Methods Twenty-eight metastatic liver cancer specimens resected from patients with colorectsl cancer, were immunohistochem-ically stained by 654-1, an anti-TP monoclonal antibody, IC6-203, another anti-TP monoclonal antibody, PG-M1, anti-macrophage marker CD68 monoclonal antibody. Morphometrical analysis and positive cell counting were performed, and the correlation of TP expression with the patient's prognosis was evaluated. Results In normal liver tissues, the hepatic cells apart from cancer nests were weakly positive for 654-1 as well as for 1C6-203. The most TP-positive cells were distributed mainly along the invasive margin of cancer or around the cancer nests. In the corresponding areas, CD68-positive macrophages were also increased. The distribution patterns of CD68-positive cells were similar to those of TP-pesitive cells. The numbers of the TP-positive cells stained by 654-1 were significantly correlated with numbers of 1C6-203 positive cells (r=0.697, P<0.01), also correlated with the numbors of CD68-positive cells (r=0.703, P<0.01). While the numbers of 1C6-203 positive cells had no significant differences with the numbers of CD68-positive cells (r=0.359, P>0.05). The TP-pesitive cancer cells both for 654-1 and for 1C6-203 were detected only in 2 of 28 specimens. Both the number of TP-pesitive cells for 654-1 and 1C6-203, and the number of CD68-positive cells had no correlation with the survival period of patients. Conclusions In the metastatic liver cancer tissues of human colorectsl cancer, the TP-expreasinn stained by 654-1 was coincidence with 1C6-203, and the most important source of TP-expreasion is the TAM in stromal tissues around cancer nests, while the cancer cells are little expressed. The numbers of TP-positive cells stained by 654-1 are significantly related with CD68-pesitive macrophages, but not with the post-operation survival period of patients.

Expression of thymidine phosphorylase in hepatocellular carcinoma and its clinical significance / 中华肝胆外科杂志

Objective To determine the significance of thymidine phosphorylase for prognosis of HCC.Methods The clinical data and pathological finding of 65 patients with HCC treated in our hospital were retrospectively analyzed.Immunohistochemical staining was performed to evaluate the expression of TP and MVD count.Statistical analysis was used to analyze the relationship between TP expression and other pathological parameters.Survival curves in 54 patients were plotted using the Kaplan-Meier method.Results The positive expressed rate of HCC tissues was significantly higher than that of normal liver tissues (P<0.01).MVD count in positive-TP group and negative-TP group was respectively 16.14±5.11,10.11±4.55.The difference between two groups had statistical significance (P<0.01).TP expression was positively correlated with tumor size,Edmondson-Steiner grade,tumor capsule status,microsatellite marker and tumor thrombi in portal vein (TTPV).The rate of postoperational recurrence after one,two and three years in TP-positive group was respectively 40%,66.7%,79.2% and it was respectively 27.4%,31.3% and 45.7% in TP-negative group.The postoperative recurrence rate after two years and three years in poitive-TP group was significantly higher than that in negative-TP group (P<0.01).In 54 patients with HCC followed up successfully,the total survival rate in TP-positive group was significantly lower than in TP-negative group (P<0.01).Conclusion TP plays an important role in neovascularization of HCC and TP may be a new prognosis index of HCC.

The Schedule Dependency for Induction of Thymidine Phosphorylase Activity and the Enhancement of Capecitabine Efficacy by Docetaxel on the SNU-484 Gastric Cancer Cell Line That was Injected into Xenografted Nude Mice

PURPOSE: Docetaxel (Taxotere(R)) and capecitabine are used in combination to treat advanced gastric cancer. Thymidine phosphorylase (TP) is an essential enzyme for the activation of capecitabine in tumors. This study sought to identify the best combination therapy with capecitabine and using two different schedules for docetaxel, a TP up-regulator, to enhance capecitabine's efficacy. METHODS: The human gastric cancer cell line SNU-484 was cultured and docetaxel (2 microgram/ml) was added to the 24-well plates that contained 5 x 10(5) cells/well. The total RNA was isolated and RT-PCR was done to identify the TP expression. Four- or five-week-old BALB/c-nu/nu mice were subcutaneously inoculated with the SNU-484 cells. The nude mice were divided into two groups and they were given capecitabine 539 mg/m2 p.o. from days 1 to 14: Group 1 was given docetaxel 15 mg/m2 i.v. on day 1; Group 2 was given docetaxel 7.5mg/m2 on days 1 and 8. Tumor tissues were excised on days 1, 8 and 15 to measure the TP and bcl-2 levels. RESULTS: TP was expressed 2 hours after docetaxel administration. Group 2 had a higher TP concentration in the tumor tissues and a better antitumor effect than did Group 1. There was no difference in the bcl-2 concentration in the two groups. CONCLUSION: These results suggest that docetaxel stimulates the TP expression in tumor tissues and it enhances the antitumor activity of capecitabine. A weekly docetaxel injection with capecitabine administration can be used to treat gastric cancer more effectively than when docetaxel is injected once per cycle. Capecitabine had no bcl-2 suppressive effect in this study.

The Expression of Thymidine Phosphorylase in Cancer-infiltrating Inflammatory Cells in Stomach Cancer

Thymidine phosphorylase (TP) has shown to be up-regulated in several cancers and to play a role in angiogenesis and invasion. Most studies regarding TP have focused on cancer cells. Recently, evidences suggest that TP in cancer-infiltrating inflammatory cells (CIICs) also affect the cancer cell behavior. To evaluate the significance of TP expression of CIICs in gastric cancer, we assessed TP expression of cancer cells and CIICs separately using immunohistochemical assay on 116 paraffin-embedded tissue samples from stomach cancer patients and investigated their clinical significance. When subjects were divided into 4 groups according to the TP expression: cancer/matrix (+/+), C/M (+/-), C/M (-/+), and C/M (-/-), intratumoral microvessel density scores were higher in the C/M (+/-) group than in the C/M (-/-) group (p=0.02). For lymph node metastasis and survival, there were no significant differences among the 4 groups. However, there were significant differences in survival (p=0.035) and LN metastasis (p=0.023) between the two groups divided by TP expression of CIICs alone irrespective of TP expression of cancer cells. Taken together, this study suggested the TP expression in CIICs could affect lymph node metastasis and patients' survival in gastric cancer.

Expression of thymidylate synthase and thymidine phosphorylase in gastric cancer and their correlation with prognosis / 中国癌症杂志

Background and purpose:Chemotherapy plays an important role in the treatment of gastric cancer.It is becoming a direction that the choice of chemotherapeutic agent for the clinic will be based on the levels of some relative gene expressions in the tumor tissue.As the key enzymes of fluoropyrimidines,the expression levels of thymidylate synthase(TS) and thymidine phosphorylase(TP) may correlate with the response of tumor to chemotherapy.This study was to investigate the expression of TS and TP mRNA in gastric cancer tissues and their correlation with prognosis.Methods:The expression levels of TS,TP mRNA in 51 gastric adenocarcinoma tissues were detected by real-time quantitative reverse transcription polymerase chain reaction(RT-PCR).Results:The median expression levels of TS,TP mRNA were 0.94 and 21.20,respectively.There was significant difference in terms of disease-free and overall survival between the gastric cancer patients with high and low TS mRNA expression level(P0.05)but not overall survival(P0.05).Conclusions:The expression levels of TS,TP mRNA may serve as prognostic markers for gastric cancer patients treated with FU-based adjuvant chemotherapy.

Determination of thymidine phosphorylase in patients with lung cancer and its clinical significance / 中国实用内科杂志

0.05).The differences between NSCLC and SCLC reached the statistical signifcance,P

Thymidylate Synthase, Thymidine Phosphorylase, VEGF and p53 Protein Expression in Primary Colorectal Cancer for Predicting Response to 5-fluorouracil-based Chemotherapy / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: In the treatment of advanced metastatic colorectal cancer, several new agents, such as irinotecan and oxaliplatin, have been developed, which have improved both disease free and overall survivals. Among these agents, 5-fluorouracil (5-FU) still remains one of the most active agents, and the selection of patients who can benefit from 5-FU-based chemotherapy is still important, as those unlikely to benefit could be spared the harmful side effects. The expression levels of thymidylate synthase (TS), thymidine phosphorylase (TP) and p53 have been known to be associated with the clinical response to 5-FU-based therapy as well as the prognosis, and that of vascular endothelial growth factor (VEGF) is associated with poor survival. MATERIALS AND METHODS: The relationship between the expressions of TS, TP, VEGF and p53 in primary tumors, using immunohistochemistry, and the response of 45 metastatic colorectal cancer patients (M: F=25: 20, median age 59 yrs) to 5-FU-based chemotherapy were evaluated. RESULTS: Thirty-seven patients were treated with 5-FU/ LV/irinotecan (FOLFIRI) and 8 with 5-FU/LV/oxaplatin (FOLFOX). The overall response rate was 28.9% (13/45). When immunohistochemically analyzed with monoclonal antibodies against TS, TP, VEGF and p53, 55.6% of the patients (25/45) were positive for TS, 48.9% (22/45) for TP, 82.2% (37/45) for VEGF, and 80% (36/45) for p53. There was a significant difference in the intensity of TS expression between the clinical responders and non-responders (p=0.036). In terms of the staining pattern of TS expression, diffuse staining was correlated with a poor response (p=0.012) and poor survival (p=0.045). However, there was no correlation between the expressions of TP, VEGF or P53 and the response to chemotherapy. CONCLUSION: These results suggest that the expression of TS in primary colorectal cancer might be an important prognostic factor for chemotherapy response and survival, and might be a useful therapeutic marker for the response of chemotherapy.

Thymidine phosphrylase (TP) in cancer-infiltrating inflammatory cells in stomach cancer: the significance of TP in prognosis and tumor angiogenesis / 대한내과학회지

BACKGROUND: Thymidine phosphorylase (TP) is an enzyme catalyzing the reversible phosphorolysis of thymidine to thymine and 2-deoxyribose-1-phosphate. TP plays a role in angiogenesis. Evidences suggest that infiltrating inflammatory cells adjacent cancer cells may affect tumor cell behavior. To evaluate each of these significances of TP expression in cancer cell and cancer-infiltrating inflammatory cells, we investigated TP expression patterns in cancer cells and infiltrating inflammatory cells adjacent cancer cells separately and the relationship between TP expression and angiogenesis or survival. METHODS: Immunohistochemistry assays were performed with anti-TP monoclonal antibody (Roche Japan) and anti-factor VIII polyclonal antibody (Dako) on 92 paraffin-embedded tissue samples from stomach cancer patients. A single pathologist scored the slides for percent positivity of tumor cells, intensity, localization and distribution of expression. TP reactivity in tumor cells (cancer) and infiltrating mononuclear cells adjacent cancer cells (matrix) was separately accessed. According to the pattern of TP expression, subjects were divided into 4 groups for further analysis: cancer(C;+)/matrix(M;+), cancer(+)/matrix(-), cancer(-)/matrix(+) and cancer(-)/matrix(-). With these 4 subsets of TP expression patterns, we evaluated cancer cell differentiation, intratumoral microvessel density, extent of tumor invasion, LN stage, and patient survival to find any differences among the subsets. RESULTS: Of 92 stomach cancer tissue, C/M(+/+), C/M(+/-), C/M(-/+), and C/M(-/-) were observed in 33patients, 19, 30, and 10, respectively. Microvessel density scores were higher in cancer(+)/matrix(-) group compared in cancer(-)/matrix(-) group (p=0.02). Of 4 TP expression subsets, other clinical factors such as histology, extent of tumor invasion, and LN metastasis were not associated with TP expression. CONCLUSION: This study suggested the TP in cancer-infiltrating inflammatory cell as well as cancer cells themselves may play an important role in angiogenesis as co-active factors in stomach cancer.

Thymidine Phosphorylase Expression in Progression of Cervical Lesions: Histological Features and Microvessel Density / 대한산부인과학회지

OBJECTIVE: The object of this study was to clarify the association of thymidine phosphorylase (TP) assessed in cancer cells and in stromal cells, with clinico-pathological factors including tumor angiogenesis and prognosis in cervical cancer. METHODS: From January 1999 to December 2001, 45 cervical tissue specimens were obtained by surgical resection in the Kyung Hee University Medical Center. The study group included 25 cases in invasive squamous cell carcinomas (SCC), 9 cases in carcinoma in situ (CIS), 7 cases in microinvasive carcinoma, 4 cases in the benign uterine diseases. They were analyzed for the cellular expression of TP and the intratumoral density of microvessels by immunohistochemistry using monoclonal antibodies to TP and factor VIII related antigen, respectively. RESULTS: Our data showed that TP expression and MVC (microvessel count) increased with histologic stage from normal, through CIS to SCC, respectively. The manifestaion of TP in the epithelium and the stroma is closely related with angiogenesis. Intraepithelial tumor revealed high expression rate of TP in the stroma, invasive cervical cancer in the epithelium, microinvasive cancer in the stroma and epithelium showing different areas of manifestation for each histologic condition, but did not show a statistically significant difference. In the case of cervical cancer, the more progressive the cancer, angiogenesis and the expression of TP increased significantly. Especially in the case of invasive cancer, stromal TP expression rate was high. CONCLUSION: These results suggest that thymidine phosphorylase might play an important role in angiogenesis, involving? cooperative epithelial and stromal expression of enzyme. Thymidine phosphorylase thus could be useful for a marker in assessing the survival rate in patients with cervical cancer.

Expression of Thymidine Phosphorylase in Human Gastric Carcinoma

PURPOSE: Angiogenesis plays an important role in the growth and metastasis of solid tumors. It has been recently reported that thymidine phosphorylase (dThdPase) is identical to platelet-derived endothelial cell growth factor. In this study, we analyzed the correlation between dThdPase activity and neovascularization, and also determined their prognostic significance by enzyme-linked immunosorbent assay (ELISA) and immunohistochemical staining in patients with gastric carcinoma. METHODS: The medical records of 57 patients with gastric carcinoma who underwent radical gastrectomy were retrospectively reviewed. Primary tumors were studied by immunohistochemical staining with anti-PD-ECGF/dThdPase and anti- CD34 monoclonal antibodies. The dThdPase activity level was also analyzed by ELISA. Microvessels were assessed by immunostaining endothelial cells for CD34. We counted the number of microvessels in the tumors of the patients. RESULTS: The dThdPase activity of tumor tissues (82.4+/-66.2 unit/mg protein) was significantly higher than that of normal mucosas (8.9 +/- 18.2 unit/mg protein) (P < 0.0001). Immunohistochemical staining with anti-dThdPase and anti-CD34 monoclonal antibodies was performed in 57 gastric carcinoma tissue samples. Positive dThdPase expression was observed in 24 (42.1%) tumors. The average number of cells expressing CD34 was significantly higher in the dThdPase- positive carcinomas (44.4+/-15.3) than in the dThdPase- negative carcinomas (30.7+/-15.8)(P<0.003). The expression of dThdPase was significantly associated with the intratu moral microvessel counts (P=00005). The patients with dThdPase-positive carcinoma showed a significantly worse prognosis than those with dThdPase-negative carcinoma (P=0.034). CONCLUSION: These results suggest that dThdPase plays a role in the promotion of angiogenesis, and that it is a possible candidate as a prognostic factor in human gastric carcinomas.

Functional Modification of a Specific RNA with Targeted Trans-Splicing

The self-splicing group I intron from Tetrahymena thermophila has been demonstrated to perform splicing reaction with its substrate RNA in the trans configuration. In this study, we explored the potential use of the trans-splicing group I ribozymes to replace a specific RNA with a new RNA that exerts any new function we want to introduce. We have chosen thymidine phosphorylase (TP) RNA as a target RNA that is known as a valid cancer prognostic factor. Cancer-specific expression of TP RNA was first evaluated with RT-PCR analysis of RNA from patients with gastric cancer. We determined next which regions of the TP RNA are accessible to ribozymes by employing an RNA mapping strategy, and found that the leader sequences upstream of the AUG start codon appeared to be particularly accessible. A specific ribozyme recognizing the most accessible sequence in the TP RNA with firefly luciferase transcript as a 3' exon was then developed. The specific trans-splicing ribozyme transferred an intended 3' exon tag sequence onto the targeted TP transcripts, resulting in a more than two fold induction of the reporter activity in the presence of TP RNA in mammalian cells, compared to the absence of the target RNA. These results suggest that the Tetrahymena ribozyme can be a potent anti-cancer agent to modify TP RNAs in tumors with a new RNA harboring anti-cancer activity.

Immunohistochemical Study on the Expression of Thymidine Phosphorylase and Ki-67 in the Uterine Cervical Neoplasia / 대한산부인과학회잡지

OBJECTIVE: We evaluated the relationship between the expression of Ki-67 and thymidine-phosphorylase (TP) according to the cancerous progression of uterine cervical cancer with immunohistochemical method. METHODS: The material was obtained from hysterectomized uterus and punched cervical specimen for two years from 1998 to 1999 at the Soonchunhyang Chunan hospital. The material included 15 normal epithelium, 13 CIN I/II, 21 CIN III, 15 microinvasive carcinoma and 13 invasive carcinoma. Monoclonal antibodies of Ki-67 and TP were used for immunohistochemical determination of cellular proliferation and angiogenic activity. RESULTS: 1. The positive rate of thymidine phosphorylase in each group of normal epithelium, CIN I/II, CIN III, microinvasive carcinoma and invasive carcinoma were 6.7%, 23.1%, 38.0%, 73.3%, 84.6% respectively. 2. The labeling indexes of Ki-67 in each group of normal epithelium. CIN I/II, CIN III, microinvasive carcinoma and invasive carcinoma were 2.0+/-0.7, 26+/-5.4, 41.2+/-10.1, 74.7+/-9.3 respectively. 3. There was statistically significant relationship between TP and Ki-67 expression. CONCLUSION: The above results indicates that the angiogenic activities and cellular proliferation indices increase according to the invasiveness of cervical cancer. We were able to reveal the expression of TP and Ki-67 and their relationship in cervical carcinoma.