结直肠癌组织中胸苷酸磷酸化酶的表达及其临床意义

背景与目的：血管生成已被公认是与肿瘤的生长和侵袭有关,近年研究表明胸苷酸磷酸化酶(thymidine phosphorylase,TP)不仅参与肿瘤新生血管形成,而且可增强5-氟尿嘧啶前药的化疗敏感性,但其在结直肠癌中的作用以及对预后的影响尚未得出一致的结论。本研究检测TP在人结直肠癌组织中的表达水平,并探讨其临床意义。方法：以西安交通大学第一医院行根治术且随访满5年的70例结直肠癌患者的库存组织标本为材料,采用免疫组织化学方法检测其TP表达水平。结果：TP表达主要见于肿瘤细胞胞浆之中,TP阳性表达患者40例,占57．14%。TP在不同Dukes’分期和不同病理组织学分级组织中的表达水平具有显著性差异(P=0．007和P=0．002)。Kaplan-Meier生存率分析提示,TP阳性表达者5年生存率为33．33％,而TP阴性表达者则为73．33％(P=0．000),结果提示TP阳性表达患者的预后较阴性表达者差。结论：TP表达可作为判断结直肠癌患者预后的参考指标之一。     

结直肠癌组织胸苷酸合成酶的表达及临床意义

目的:探讨了胸苷酸合成酶(TS)在结直肠癌组织中的表达及其与临床病理特征和预后的关系。方法:用免疫组化SP法检测72例结直肠癌组织中TS的表达,按TS表达水平将患者分为2组:高表达组和低表达组,用Kaplan-Meier法分析TS的表达与临床病理特征和预后的相关性。用多因素COX模型分析影响患者预后的相关因素。结果:在72例结直肠癌组织中TS表达从15%~95%不等,平均(49.79±25.05)%。TS的表达随Dukes分期的增加而增加,但无统计学意义(P=0.145)。早期(Dukes A~B)结直肠癌的预后明显优于晚期结直肠癌(Dukes C、D),P<0.0001。TS低表达者预后优于高表达患者。治疗前血清CEA、CA19-9水平正常者预后较好,CEA、CA19-9升高者预后较差。多因素分析显示Dukes分期、TS表达状况、CEA是影响预后的因素。结论:结直肠癌治疗前Dukes分期是影响预后的主要因素。TS的表达状况、CEA也是影响预后的参考指标。     

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。     

胸苷酸合成酶在膀胱癌组织中的表达及意义

目的探讨膀胱癌（BC）中胸苷酸合成酶（TS）的表达对膀胱癌患者预后的影响。方法采用免疫组化S-P法检测50例膀胱癌石蜡标本中的TS蛋白表达情况。结果TS蛋白在膀胱癌与正常膀胱黏膜中的表达有显著性差异（P＜0．01），与肿瘤低分化、临床病理分期晚期、淋巴结转移密切相关（P＜0．05）。结论TS在膀胱癌中的表达增强可能是膀胱癌患者一个重要的预后指标。     

胸苷酸合成酶、胸苷酸磷酸化酶的表达与培美曲塞时辰化疗的疗效分析

目的构建肺腺癌A549细胞的裸鼠皮下移植瘤模型,研究按昼夜不同时辰给予培美曲塞是否能提高疗效和降低不良反应,分析胸苷酸合成酶（TS）mRNA和胸苷酸磷酸化酶（TP）mRNA表达高低与培美曲塞疗效以及不良反应的关系。方法 常规方法体外培养肺腺癌A549细胞。待肿瘤体积生长至0.5~1.5 cm³,随机将荷瘤鼠分为四组,每组5只,其中三组严格按照每昼夜24 h的三个时间点给药即:早晨7点（光照后0 h）、中午15点（光照后8 h）、晚间23点（光照后16 h）进行腹腔给药,第四组为对照组,腹腔给予同体积的0.9%氯化钠溶液。培美曲塞按照150 mg/kg应用,每天一次,连续4天。以后每周测量瘤体积,裸鼠体重,观察裸鼠的行为状态、精神以及食欲变化。实验结束时称量剥离的裸瘤重量。应用实时荧光定量PCR（Real-Time PCR）检测肿瘤组织中TS mRNA和TPmRNA的表达水平。结果7 h给药组裸鼠体重增长较快（P=0.041）,化疗不良反应较小,瘤体积生长最慢（P=0.032）,抑瘤率最大（P=0.000）。7 h给药组的肿瘤组织中TS mRNA相对表达量最低,TP mRNA的相对表达量对高,差异有统计学意义（P<0.01）。结论培美曲塞对肺腺癌裸鼠移植瘤有明显的抑制作用,疗效及不良反应与时辰给药有关,7 h给药组疗效最佳,化疗不良反应最小;TS和TP可以作为培美曲塞的疗效预测因子,为培美曲塞的个体化临床用药提供参考。     

胸苷酸磷酸化酶在胃肠肿瘤中的研究进展

胃肠肿瘤是最常见的恶性肿瘤，在我国发病率呈明显上升趋势。5-Fu是治疗胃肠肿瘤最基本的化疗药物之一，胸苷酸磷酸化酶（thymidine phosphorylase，TP）是5-Fu在体内转化为具有抗癌活性的关键性靶酶，近来研究发现TP不仅参与肿瘤血管的生成，而且有助于预测肿瘤患者对5-Fu的化疗毒性和敏感性，TP表达水平在一定程度上可反映胃肠肿瘤的恶性度高低，现将TP与胃肠肿瘤的关系综述如下。     

胸苷酸合成酶在膀胱癌组织中的表达及意义

目的探讨膀胱癌(BC)中胸苷酸合成酶(TS)的表达对膀胱癌患者预后的影响。方法采用免疫组化S_P法检测50例膀胱癌石蜡标本中的TS蛋白表达情况。结果TS蛋白在膀胱癌与正常膀胱黏膜中的表达有显著性差异(P<0.01),与肿瘤低分化、临床病理分期晚期、淋巴结转移密切相关(P<0.05)。结论TS在膀胱癌中的表达增强可能是膀胱癌患者一个重要的预后指标。     

胸苷磷酸化酶在肿瘤组织中的表达及其临床应用

胸苷磷酸化酶（TP）参与嘧啶核苷合成和分解的过程,作为一种血管生成因子,在肿瘤的生长、浸润和转移过程中有重要的作用,也在5-Fu等药物的代谢中起作用.现综述TP可能的促癌机制、TP在肿瘤组织中的表达及与临床病理因素的关系、TP的临床应用价值.     

胃癌组织中胸苷酸合成酶mRNA表达与预后的关系

目的探讨胃癌组织中胸苷酸合成酶（TS）mRNA表达水平与预后的关系。方法以G6PDH作内参照,采用实时定量RT-PCR技术检测41例胃癌组织TS mRNA表达水平。结果胃癌组织TS mRNA表达水平的中位数为0．93。TS高表达组（〉0．93）和低表达组（≤0．93）之间无瘤生存期和总生存期差异有显著性（P〈0．05）;而TS mRNA表达与年龄、性别、淋巴结转移、组织学分级及临床分期均无相关性（P〉0．05）。结论检测TS mRNA表达水平对判断胃癌病人预后有很好的预测价值。     

Expression of thymidylate synthase and thymidine phosphorylase in recurrence and survival rates of advanced gastric cancer

Background. The immunohistochemical expression of thymidylate synthase (TS) and thymidine phosphorylase (TP) was examined in a comparative study of the recurrence rates and prognoses of patients with advanced gastric cancer at the same stage. Methods. We examined the resected specimens of 67 patients with stage IIIB gastric cancer (pT3, pN2, M0) under 70 years of age who had undergone curative gastrectomy followed by adjuvant chemotherapy with 5-fluoropyrimidines. Paraffin sections of the resected specimens were stained with human anti-TS polyclonal and anti-TP monoclonal antibodies by the avidin-biotin-peroxidase complex (ABC) method. Results. The overall expression of TS and TP was 45.4% and 43.4%, respectively. The postoperative survival curve for the TS-positive group was significantly depressed compared with that for the TS-negative group ( P = 0.0480). The survival curves for TP-positive and TP-negative groups did not show any difference. In regard to the combination of TS and TP expression, the best survival curve was obtained for the TS(−)/TP(+) group, followed by those for the TS(−)/TP(−), TS(+)/TP(−), and TS(+)/TP(+) groups in descending order. With regard to the recurrence site, there was no significant difference in peritoneal recurrence in relation to positivity for TS or TP. Lymph node recurrence, however, was significantly higher in the TS-positive and TP-positive groups, with P -values being 0.0466 and 0.0058, respectively, versus the corresponding negative groups. The incidence of hepatic recurrence was higher in the TP-positive group than in the TP-negative group ( P = 0.0910). As for the total doses of 5-fluoropyrimidines given, more favorable survival curves were obtained for the high dose of negative TS and TP groups, but no significant differences were observed in their positivities. Conclusion. The expressions of TS and TP showed different characteristics in overall survival and recurrence rate or site. They should be used for predicting prognosis in comprehension on their properties. Received for publication on Dec. 21, 1998; accepted on Sept. 1, 1999     

乳腺癌组织中TP和TS及DPD mRNA表达与预后的关系

目的　探讨乳腺癌组织中胸苷酸化酶 (TP)、胸苷酸合成酶 (TS)和二氢嘧啶脱氢酶(DPD)mRNA表达水平及其与预后的关系。方法　采用real time定量PCR技术检测经过微选的 9例正常乳腺组织和 86例乳腺癌组织TP、TS和DPD的mRNA表达水平。结果　肿瘤组织中TP、TS和DPDmRNA表达水平中位数分别为 16 .5 4 ,0 .38和 2 .74 ,正常乳腺组织分别为 11.75 ,0 .2 5和 8.33,差异均无显著性 (P >0 .0 5 )。除年龄与DPD表达呈负相关外 ,TP、TS和DPDmRNA表达与肿瘤体积、淋巴结转移、组织学分级、临床分期均无相关性。TP、DPD高表达组和低表达组之间 ,无瘤生存期和总生存期差异均无显著性。TS高表达组和低表达组无瘤生存期差异无显著性 (P =0 .0 6 9) ;平均总生存期分别为 5 9.0 0和 70 .30个月 ,差异有显著性 (P =0 .0 4 96 )。结论　仅检测TSmRNA对判断乳腺癌预后有参考价值 ,同时检测TP、TS和DPD具有更好的预测价值。     

Association of thymidylate synthase polymorphisms with gastric cancer susceptibility

We investigated in a case-control study a possible role of thymidylate synthase gene (TS) polymorphisms for gastric cancer susceptibility. Lymphocyte genomic DNA from 134 Italian gastric cancer patients and 139 controls was used for genotyping two polymorphisms in the TS 5'-untranslated region (5'-UTR); a double (2R) or triple (3R) 28-bp repeat and a G/C polymorphism within the triple repeats allele (3G allele). Samples were also genotyped at a 6-bp deletion/insertion (del6 or ins6) polymorphism at position 1494 in the TS 3'-untranslated region (3'-UTR). Unconditional regression with odd ratios (OR) and 95% confidence intervals (CI), haplotype and linkage disequilibrium analyses were used to investigate the association of the polymorphisms with the disease. The global allelic distribution was in Hardy-Weinberg equilibrium. Genotypes with the 3G allele (2R/3G, 3C/3G, 3G/3G) were significantly more frequent in patients than controls and were associated with gastric cancer risk (OR = 2.06; 95% CI = 1.26-3.35). A significant risk was also observed for carriers of the del6 allele in the 3'-UTR. Odds ratios for combined 3G-del6/ins6 and 3G-del6/del6 genotypes were 2.59 (95% CI = 1.36-4.94) and 2.81 (95% CI = 1.22-6.64), respectively. The 3G-del6 haplotype showed a significant association with the disease (p = 0.01). Polymorphisms in the TS gene may contribute to gastric cancer susceptibility and this finding deserve further investigation in the context of novel strategies for gastric cancer prevention. In vitro, 3G genotypes have been related to high TS mRNA expression, which may underlie one of the possible etiologic mechanisms.     

Expression of thymidylate synthase in gastric cancer patients treated with 5-fluorouracil and doxorubicin-based adjuvant chemotherapy after curative resection

We evaluated the expression of thymidylate synthase (TS) in locally advanced gastric cancer patients treated with adjuvant chemotherapy after curative resection and investigated the association between TS expression and clinicopathologic characteristics including prognosis of the patients. TS expression was evaluated by immunohistochemical staining using TS106 monoclonal antibody in 103 locally advanced gastric cancer patients (stage IB-IV) who underwent 5-fluorouracil (5-FU) and doxorubicin-based adjuvant chemotherapy after curative resection. 65 patients (63%) had primary tumours with high TS expression (> or = 25% of tumour cells positive), and 38 patients (37%) demonstrated low TS expression (< 25% of tumour cells positive or no staining). High TS expression was associated with male gender (P = 0.002), poorly differentiated histology (P = 0.015), and mixed type in Lauren's classification (P = 0.027). There were no statistically significant differences in 4-year disease-free survival (60.0% vs. 57.2%, P = 0.548) and overall survival (59.6% vs. 59.3%, P = 0.792) between high-TS group and low-TS group. In conclusion, although high TS expression was associated with poorly differentiated histology and mixed type in Lauren's classification, it did not predict poor disease-free and overall survival in gastric cancer patients treated with 5-FU and doxorubicin-based adjuvant chemotherapy after curative resection. Further prospective studies including the evaluation of other biological markers associated with the resistance to 5-FU and doxorubicin are necessary.     

Vorinostat synergises with capecitabine through upregulation of thymidine phosphorylase

Background:

Potentiation of anticancer activity of capecitabine is required to improve its therapeutic index. In colorectal cancer (CRC) cells, we evaluated whether the histone deacetylase-inhibitor vorinostat may induce synergistic antitumour effects in combination with capecitabine by modulating the expression of thymidine phosphorylase (TP), a key enzyme in the conversion of capecitabine to 5-florouracil (5-FU), and thymidylate synthase (TS), the target of 5-FU.

Methods:

Expression of TP and TS was measured by real-time PCR, western blotting and immunohistochemistry. Knockdown of TP was performed by specific small interfering RNA. Antitumour activity of vorinostat was assessed in vitro in combination with the capecitabine active metabolite deoxy-5-fluorouridine (5′-DFUR) according to the Chou and Talay method and by evaluating apoptosis as well as in xenografts-bearing nude mice in combination with capecitabine.

Results:

Vorinostat induced both in vitro and in vivo upregulation of TP as well as downregulation of TS in cancer cells, but not in ex vivo treated peripheral blood lymphocytes. Combined treatment with vorinostat and 5′-DFUR resulted in a synergistic antiproliferative effect and increased apoptotic cell death in vitro. This latter effect was impaired in cells where TP was knocked. In vivo, vorinostat plus capecitabine potently inhibited tumour growth, increased apoptosis and prolonged survival compared with control or single-agent treatments.

Conclusions:

Overall, this study suggests that the combination of vorinostat and capecitabine is an innovative antitumour strategy and warrants further clinical evaluation for the treatment of CRC.     

TP/PD-ECGF表达与血小板增多在胃癌中的意义

目的：研究血小板衍化内皮细胞生长因子（TP/PD-ECGF）在胃癌中的表达及血小板增多情况,并对其与胃癌患者临床病理特征和预后的关系进行探讨。方法：采用免疫组化EnVision两步法检测107例胃癌组织中TP/PD-ECGF的表达,记录血小板增多情况,并分析二者与胃癌患者临床病理特征及预后的关系。结果：胃癌患者中TP/PD-ECGF阳性表达率为71.0%,与血小板增多呈正相关（P〈0.01）。TP/PD-ECGF表达、血小板增多与肿瘤分期、淋巴结转移、远处转移及分化程度呈正相关。TP/PD-ECGF阳性与阴性表达患者3年、5年总生存率分别为69.04%、18.12%和88.87%、75.20%,二者差异有统计学意义（P=0.0383）;3年、5年无进展生存率分别为64.87%、17.92%和82.73%、35.00%,二者差异有统计学意义（P=0.0350）。Cox比例风险模型多因素分析显示,肿瘤分期、淋巴结转移、远处转移、TP/PD-ECGF及血小板增多均是影响胃癌预后独立的危险因素。结论：TP/PD-ECGF表达与血小板增多呈正相关,二者与胃癌生长和浸润转移关系密切,可作为胃癌独立的预后因素。     

TP/PD-ECGF表达与血小板增多在胃癌中的意义

目的:研究血小板衍化内皮细胞生长因子(TP/PD-ECGF)在胃癌中的表达及血小板增多情况,并对其与胃癌患者临床病理特征和预后的关系进行探讨。方法:采用免疫组化EnVision两步法检测107例胃癌组织中TP/PD-ECGF的表达,记录血小板增多情况,并分析二者与胃癌患者临床病理特征及预后的关系。结果:胃癌患者中TP/PD-ECGF阳性表达率为71.0%,与血小板增多呈正相关(P<0.01)。TP/PD-ECGF表达、血小板增多与肿瘤分期、淋巴结转移、远处转移及分化程度呈正相关。TP/PD-ECGF阳性与阴性表达患者3年、5年总生存率分别为69.04%、18.12%和88.87%、75.20%,二者差异有统计学意义(P=0.0383);3年、5年无进展生存率分别为64.87%、17.92%和82.73%、35.00%,二者差异有统计学意义(P=0.0350)。Cox比例风险模型多因素分析显示,肿瘤分期、淋巴结转移、远处转移、TP/PD-ECGF及血小板增多均是影响胃癌预后独立的危险因素。结论:TP/PD-ECGF表达与血小板增多呈正相关,二者与胃癌生长和浸润转移关系密切,可作为胃癌独立的预后因素。     

胃癌患者血清DPD mRNA和TS mRNA表达对替吉奥疗效预测价值分析

目的 探讨进展期胃癌(advanced gastric carcinoma,AGC)患者二氢嘧啶脱氢酶(thymidine phosphorylase,DPD) mRNA和胸苷酸合成酶(thymidylate synthase,TS) mRNA表达与含替吉奥(S-1)方案化疗疗效的关系.方法 收集2012-09-01-2013-09-01苏州市相城人民医院收治经病理确诊的AGC患者40例,化疗前抽取外周血采用RT-PCR法检测DPD mRNA及TS mRNA表达水平,给予含S-1的方案:S-1(40 mg/m2,2次/d,口服14 d)+顺铂(DDP,75 mg/m2,静脉滴入,d1)化疗.分析DPD mRNA及TS mRNA表达与化疗疗效的关系.结果 RT-PCR检测结果显示,40例患者的DPD mRNA和TS mRNA表达相对于β-actin的相对数值分别为2.53±1.86和0.636±0.363.DPDmRNA和TS mRNA表达水平之间有显著相关性,r=0.68,P＜0.01.AGC患者外周血中DPD mRNA表达与Lauren分型显著相关,x2 =3.259,P=0.032.TS mRNA表达则与是否有远处转移显著相关,x2 =3.294,P=0.040. DPDmR-NA及TS mRNA均是表达越高患者疗效越差,表达越低患者疗效越好,r=0.708,P＜0.01;r=0.728,P＜0.01.联合分析显示,DPD mRNA及TS mRNA同时低表达时疗效更好,x2 =13.23,P=0.004.结论 DPD mRNA及TS mRNA表达可单独或联合用于预测AGC患者对含S-1方案的化疗疗效,初步研究结果提示可以根据患者外周血DPD mRNA及TS mRNA表达来指导胃癌的个体化治疗.     

Expression of multidrug resistance-associated protein1,P-glycoprotein,and thymidylate synthase in gastric cancer patients treated with 5-fluorouracil and doxorubicin-based adjuvant chemotherapy after curative resection

Both 5-fluorouracil and doxorubicin are commonly used agents in chemotherapy of gastric cancer in adjuvant setting as well as metastatic disease. In a variety of malignancies, high expression of multidrug resistance-associated protein1 and P-glycoprotein has been associated with resistance to doxorubicin, whereas 5-fluorouracil resistance has correlated with the level of thymidylate synthase expression. We evaluated the expression of multidrug resistance-associated protein1, P-glycoprotein, and thymidylate synthase using immunohistochemistry in 103 locally advanced gastric cancer patients (stage IB-IV) who underwent 5-fluorouracil and doxorubicin-based adjuvant chemotherapy after curative resection and investigated the association between their expression and clinicopathologic characteristics including prognosis of the patients. While high expression (> or =5% of tumour cells positive) of multidrug resistance-associated protein1 and P-glycoprotein was observed in 70 patients (68%) and 42 patients (41%), respectively, 65 patients (63%) had primary tumours with high expression (> or =25% of tumour cells positive) of thymidylate synthase. There was a significant association between multidrug resistance-associated protein1 and P-glycoprotein expression (P<0.0001) as well as P-glycoprotein and thymidylate synthase expression (P<0.0001). High multidrug resistance-associated protein1 and P-glycoprotein expressions were associated with well and moderately differentiated histology (P<0.0001 and P=0.03, respectively) and intestinal type (P<0.0001 and P=0.009, respectively). High multidrug resistance-associated protein1 expression correlated with lymph node metastasis (P=0.037), advanced stage (P=0.015), and older age (P=0.021). Five-year disease-free survival and overall survival of total patients were 55.2% and 56.2%, respectively, with a median follow-up of 68 months. There were no significant differences in disease-free survival and overall survival according to the expression of multidrug resistance-associated protein1 (P=0.902 and P=0.975, respectively), P-glycoprotein (P=0.987 and P=0.955, respectively), and thymidylate synthase (P=0.604 and P=0.802, respectively). Concurrent high expression of these proteins (high multidrug resistance-associated protein1/P-glycoprotein, high multidrug resistance-associated protein1/thymidylate synthase, high P-glycoprotein/thymidylate synthase) did not correlate with disease-free survival or overall survival. Even high expression of all three proteins was not associated with poor disease-free survival (P=0.919) and overall survival (P=0.852). In conclusion, high expression of multidrug resistance-associated protein1, P-glycoprotein, and thymidylate synthase did not predict poor prognosis of gastric cancer patients treated with 5-fluorouracil and doxorubicin-based adjuvant chemotherapy. A larger study including patients treated with surgical resection alone would be necessary.