胃癌组织中胸苷酸合成酶、胸苷磷酸化酶mRNA表达及其临床意义

背景与目的:化学治疗在胃癌治疗中扮演了重要的作用。通过肿瘤组织中某些基因表达的水平来选择化疗药物已成为今后化疗的方向。胸苷酸合成酶(TS)、胸苷磷酸化酶(TP)是氟尿嘧啶(5-FU)体内的关键代谢酶,其表达水平影响恶性肿瘤患者接受5-FU化疗后的预后。本研究探讨胃癌组织中TS、TP mRNA表达水平与预后的关系。方法:采用实时定量RT-PCR技术检测51例胃腺癌组织TS、TP mRNA表达水平。结果:胃癌组织TS、TP mRNA表达水平的中位数分别为0.94和21.20,TS高表达组和低表达组之间无瘤生存期和总生存期差异有显著性(P<0.05),TP高表达组和低表达组之间总生存期差异有显著性(P<0.05),但无瘤生存期之间差异无显著性(P>0.05)。TS、TP 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、TP mRNA表达水平对接受5-FU为基础治疗方案的胃癌患者的预后有很好的预测价值。     

进展期胃癌术后化疗效果与5-氟尿嘧啶通道基因的关系

目的研究进展期胃癌术后化疗效果与5-氟尿嘧啶（5-Fu）通道基因的关系。方法收集52例进展期胃癌术后辅助FOLFOX4化疗6个周期病例，建立数据库。选择5-Fu通道基因中的胸苷酸合成酶（TS）、二氢嘧啶脱氢酶（DPD）、乳清酸磷酸核糖转移酶（OPRT）3个基因，采用反转录聚合酶链反应（RT-PCR）方法检测52例胃癌组织和临近正常组织TS、DPD、OPRTmRNA表达，探讨5．Fu通道基因对化疗效果的影响。结果肿瘤组织中相对于GAPDH，TSmRNA相对表达量为0．92±0．28，较对应正常组织中的0．71±0．30高（t=3．730，P=0．001），胃癌组织和正常组织中OPRTmRNA呈正相关（r=0．45，P=0．001），Ts和DPDmRNA表达分别在肿瘤和正常组织中无相关性；肿瘤组织和正常组织中OPRT mRNA高表达者有较长生存期（t=3．036，P=0．003；t=2．713，P=0．009），肿瘤组织低DPD mRNA表达者有较长生存期（t=2．708，P=0．009），在正常组织中，高DPDmRNA表达者有较长生存期（t=2．616，P=0．012）。结论进展期胃癌术后化疗效果与DPD和OPRT相关，与TS的表达无关。     

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。     

氟尿嘧啶代谢酶与胃肠道肿瘤化疗敏感性关系的探讨

目的探讨氟尿嘧啶类药物代谢酶--胸苷酸合成酶(TS)、胸苷磷酸化酶(TP)和二氢嘧啶脱氢酶(DPD)表达水平与人胃肠道肿瘤细胞化疗敏感性的关系.方法MTT法分析7株人胃肠道肿瘤细胞对5-FU和FdUrd的敏感性(半数抑制浓度,IC50),RT-PCR检测3种药物代谢酶mRNA的表达,ELISA测定DPD和TP蛋白含量.结果5-FU与FdUrd对肿瘤细胞的IC50值分别为1.28～12.26μmol/L和5.02～24.21 μmol/L,DPD mRNA水平和蛋白含量与5-FU的化疗敏感性呈负相关,TSmRNA水平与FdUrd敏感性呈负相关;DPD的mRNA和蛋白表达水平之间显著相关,但TP没有相关性.结论DPD和TS表达水平分别可以作为5-FU与FdUrd化疗敏感性的预测指标.     

消化道肿瘤病人外周血TSmRNA和DPDmRNA的表达

[目的]探讨胸苷酸合成酶（thymidylate synthase，TS）mRNA和二氢嘧啶脱氢酶（dihydropyrimidine dehydrogenase，DPD）mRNA在癌组织和外周血中的表达及其相互关系。[方法]采用RT—PCR检测36例胃肠道癌及外周血中TSmRNA和DPDmRNA的表达水平，同时检测32例对照组外周血中表达水平。[结果]在癌组织中TSmRNA检出52．8％（19／36），DPDmRNA检出44．4％（12／36），高于外周血的检出率30．5％（11／36）和25．0％（9／36），但差异无统计学意义fD0．05）。肿瘤组织TSmRNA、DPDmRNA和外周血的表达结果相关良好，r值分别为0．627和0．645，在外周血中肿瘤组的表达高于对照组（P〈0．01）。[结论]外周血和肿瘤组织中的TSmRNA和DPDmRNA高度相关，用RT—PCR方法检测外周血TSmRNA和DPDmR—NA，操作简单、可反复检测，适于临床应用。     

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.     

二氢嘧啶脱氢酶(DPD)与胸苷酸合成酶(TS)在结直肠癌中的表达及预后价值

目的:探讨5-FU代谢酶在结直肠癌中的表达及其与预后的关系.方法:44例结直肠癌根治术后分别施以5-FU为主的辅助化疗,并通过免疫组化检测二氢嘧啶脱氢酶(DPD)和胸苷酸合成酶(TS)的表达.结果:DPD阳性表达的结直肠癌无病生存期显著缩短(P=0.047),而总生存期也有缩短的趋势,但差异无显著意义(P=0.136).而TS表达与预后无关(P＞0.05),但TS在晚期肿瘤中表达较高(P＜0.05).结论:在接受5-FU为主辅助化疗的结直肠癌患者中,DPD表达可作为预后的重要指标.TS表达与临床分期密切相关,可视为结直肠癌进展的生物学标志.     

E2F-1和Rb基因表达与乳腺乳头状瘤病及导管内癌的相关性

目的　研究细胞周期调控因子E2F 1和Rb与乳腺乳头状瘤病及导管内癌的相关性 ,探讨乳头状瘤病癌变的某些分子机制。方法　采用原位杂交法和免疫组化法对 4 0例乳腺轻度乳头状瘤病、4 0例重度乳头状瘤病和 4 0例导管内癌进行了检测 ,观察E2F 1和Rb基因mRNA和蛋白的表达情况。结果　E2F 1mRNA表达阳性率在乳腺轻度乳头状瘤病、重度乳头状瘤病和导管内癌中分别为17.5 %、4 5 .0 %和 80 .0 % ;蛋白表达阳性率分别为 2 0 .0 %、4 7.5 %和 77.5 %。E2F 1mRNA和蛋白表达三组间差异有显著性 (P <0 .0 1) ,组间两两比较 ,差异也有显著性 (P <0 .0 1)。Rb基因mRNA表达阳性率在乳腺轻度乳头状瘤病、重度乳头状瘤病和导管内癌中分别为 90 .0 %、5 0 .0 %和 2 0 .0 % ;蛋白表达阳性率分别为 85 .0 %、5 2 .5 %和 2 2 .5 %。Rb基因mRNA和蛋白表达三组间差异有显著性 (P <0 .0 1) ,组间两两比较 ,差异也有显著性 (P <0 .0 1)。随着乳腺乳头状瘤病程度的加重最终进展为癌 ,E2F 1mRNA和蛋白表达阳性率呈上升趋势 ;RbmRNA和蛋白表达阳性率呈下降趋势。E2F 1和RbmRNA表达均与蛋白表达呈正相关 ,但这两种调控因子的表达呈负相关。结论　E2F 1和Rb基因mRNA和蛋白表达可作为癌症早期较有价值的参考指标 ,有助于从乳腺乳头状瘤病     

二氢嘧啶脱氢酶（DPD）与胸苷酸合成酶（TS）在结直肠癌中的表达及预后价值

目的：探讨5-FU代谢酶在结直肠癌中的表达及其与预后的关系。方法：44例结直肠癌根治术后分别施以5-FU为主的辅助化疗,并通过免疫组化检测二氢嘧啶脱氢酶（DPD）和胸苷酸合成酶（TS）的表达。结果：DPD阳性表达的结直肠癌无病生存期显著缩短（P=0．047）,而总生存期也有缩短的趋势,但差异无显著意义（P=0．136）。而偈表达与预后无关（P〉0．05）,但TS在晚期肿瘤中表达较高（P〈0．05）。结论：在接受5-FU为主辅助化疗的结直肠癌患者中,DPD表达可作为预后的重要指标。TS表达与临床分期密切相关,可视为结直肠癌进展的生物学标志。     

乳腺癌hMAM mRNA和CD44V6 mRNA表达及临床意义

目的研究人乳腺珠蛋白(human mammoglobin,hMAM)mRNA及CD44V6 mRNA在乳腺癌中的表达,探讨它们作为乳腺癌微转移标志物的可能性.方法采用巢式逆转录聚合酶链反应(nested-RT-PCR)技术,检测乳腺癌组织及癌旁正常乳腺组织、乳腺良性肿瘤中hMAM mRNA及CD44V6 mRNA,检测乳腺癌、乳腺良性肿瘤患者及健康志愿者外周血hMAM mRNA和CD44V6 mRNA,同时对胃癌、结直肠癌、食管癌、肺癌、卵巢癌组织中hMAM mRNA的表达进行检测.结果 hMAM-mRNA在胃癌、结直肠癌、食管癌、肺癌、卵巢癌组织中无表达,在乳腺癌、癌旁组织及乳腺良性肿瘤组织中的阳性表达率分别为96.72%、96.72%和93.75%,3组间差异无显著性(P>0.05);CD44V6 mRNA在乳腺癌组织中的阳性表达率明显高于癌旁组织及乳腺良性肿瘤组织(P<0.05).乳腺癌患者外周血hMAM mRNA检出率为45.90%,Ⅳ期乳腺癌患者外周血中hMAM mRNA的阳性表达率明显高于其他各组(P<0.05);而hMAM mRNA在健康人及乳腺良性肿瘤患者外周血中均无表达;CD44V6 mRNA在乳腺癌、乳腺良性肿瘤患者和健康人外周血中的阳性表达率各组之间无显著性差异(P>0.05).结论 hMAM mRNA相对特异性地表达于乳腺组织,外周血检测出hMAM mRNA提示已有癌细胞进入血流,因此可能作为检测乳腺癌血道微转移的标志物;而CD44V6 mRNA作为血液中乳腺癌细胞的特异性标志物缺乏特异性.     

Combined GADD45A and thymidine phosphorylase expression levels predict response and survival of neoadjuvant-treated gastric cancer patients.

PURPOSE: We evaluated the expression of seven therapy-related genes to predict the clinical outcome of advanced gastric cancer patients treated with a neoadjuvant chemotherapeutic protocol. EXPERIMENTAL DESIGN: Pretherapeutic, formalin-fixed, and paraffin-embedded biopsies of 61 patients, who received a 5-fluorouracil (5-FU)- and cisplatin-based chemotherapy were studied. The expressions of the 5-FU-related genes TS, DPD, and TP and of the cisplatin-related genes ERCC1, ERCC4, KU80, and GADD45A were analyzed by quantitative real-time PCR. The expression levels of single genes and of various combinations were tested for an association with response and overall survival. RESULTS: High DPD levels were more frequently found in nonresponding patients and were associated with worse survival. GADD45A and TP levels showed weak associations with response, but GADD45A expression correlated with survival. There was no association with response for TS expression, but tumors with a high TS level were associated with worse survival. The combination of GADD45A and TP revealed the strongest predictive effect. High expression values of TP and/or GADD45A were exclusively found in nonresponding patients (P = 0.002) and were associated with a significantly poorer survival (P = 0.04). CONCLUSIONS: Combined gene expression levels of TP and GADD45A represent a new variable to predict the clinical outcome after neoadjuvant chemotherapy in gastric cancer. The association of DPD expression with response and survival underlines a predominant role of DPD to predict 5-FU sensitivity. The association of TS expression levels with survival but not with response suggests an importance of this gene for tumor progression.     

Thymidylate synthetase (TS) genotype and TS/dihydropyrimidine dehydrogenase mRNA level as an indicator in determining chemosensitivity to 5-fluorouracil in advanced gastric carcinoma

BACKGROUND: One of the target enzymes of 5-fluorouracil (5-FU) is thymidylate synthetase (TS). The DNA sequence of the TS gene includes either double or triple tandem 28-bp repeats within the promoter region, such that TS genotypes can be classified as homozygous 3R/3R heterozygous 2R/3R or homozygous 2R/2R Several recent studies have shown that TS genotype affects mRNA expression, with 3R/3R homozygotes showing higher TS mRNA expression compared to the other genotypes. PURPOSE: We analyzed the TS genotype and TS and dihydropyrimidine dehydrogenase (DPD) mRNA expression levels in 22 advanced gastric carcinoma patients, and analyzed results with respect to patient 5-FU chemosensitivity, as detected by the tetrazolium-based colorimetric (MTT) assay and survival outcome. PATIENTS AND METHODS: Between September 2001 and April 2002, 22 Japanese patients with advanced gastric carcinoma were evaluated. Informed written consent was obtained from all patients and the study was approved by the ethical committee at our University Hospital Fresh surgical specimens from carcinoma lesions were enzymatically dissociated and incubated with 5-FU at a concentration of 50 microg/ml for 48 hours to determine the inhibition rate as detected by MTT assay. Normal and tumor tissue and peripheral blood samples were collected and stored at -80 degrees C until assay for TS genotype and TS and DPD mRNA level The TS genotype was assessed by PCR assay using peripheral monocytes, since monocyte genotypes represent the genotype of normal and tumor tissues. Quantification of TS and DPD mRNA levels was performed using real-time PCRP Survival outcome was assessed according to the disease-free survival period in cases with similar clinical backgrounds. RESULTS: TS genotyping revealed 19 3R/3R homozygotes and 3 2R/3R heterozygotes. After analysis of normal and tumor tissues, samples from homozygote 3R/3R cases showed higher TS mRNA expression than heterozygote 2R/3R cases, which was statistically significant at p<0.05. We also observed a statistically significant correlation in TS mRNA levels between normal and tumor tissues, while no significant correlation was observed for DPD mRNA levels between normal and tumor tissues. While no relationship between 5-FU chemosensitivity and TS genotype or mRNA expression was observed, cases with high DPD mRNA expression were resistant to 5-FU and exhibited poor survival outcomes. CONCLUSION: While TS genotype affected TS mRNA expression in both normal and tumor tissues in advanced gastric cancer, there is no relationship between TS genotype or mRNA expression level and 5-FU chemosensitivity. CONCLUSION: Our finding, that DPD mRNA expression appears to be a factor in determining 5-FU chemosensitivity and the survival outcome of advanced gastric cancer patients, is comparable with previous reports.     

A retrospective study on TS mRNA expression and prediction of the effects of adjuvant oral 5-fluorouracil in breast cancer

Nucleic acid-metabolizing enzymes, such as thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP) and orotate phosphoribosyl transferase (OPRT), have attracted attention as candidates for response determinants of 5-fluorouracil (5-FU). Whether the expression levels of these enzymes can be adopted as valuable parameters for 5-FU sensitivity in breast cancer has yet to be elucidated. In the present study, intratumoral mRNA expression of TS, DPD, TP and OPRT were determined in formalin-fixed paraffin-embedded surgical specimens collected from 217 breast cancer patients, using the Danenberg Tumor Profile method, which combines microdissection and real-time-polymerase chain reaction. The significance of these enzymes as prognostic and 5-FU efficacy-predicting factors was evaluated. Our data showed that a low DPD expression is related to a high nuclear grade and other factors including hormone receptor-negativity. Low expression levels of TP were found in hormone receptor-negative tumors. TS and OPRT expression were not related to various clinicopathological factors, but patients with a high TS mRNA expression showed a significantly poorer prognosis in cases where 5-FU was not administered. The efficacy of 5-FU was more significant when administered for more than 6 months in the group with a high TS mRNA expression. These data suggest that TS mRNA expression in breast cancer tissue is an ideal predictor of outcomes for patients with no administration of 5-FU, and of the efficacy of 5-FU.     

Possibility of using mRNA expression levels for nucleic acid-metabolizing enzymes within prostate cancer cells as indices for prognostic factors

Thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP) and orotate phosphoribosyl transferase (OPRT) are enzymes involved in nucleic acid metabolism. It has been reported (based on observations of various tumor types) that the extent of the mRNA expression of these enzymes within tumor tissues may be used as a factor to define tumor prognosis. It has also been reported that the mRNA expression patterns differ in each type of tumor. However, few reports are available on the distribution of mRNA expression in prostate cancers. This study was conducted on tissue specimens obtained from 172 patients who were diagnosed with prostate cancer and had undergone total prostatectomies. The mRNA expression of TS, DPD, OPRT and TP was quantitatively analyzed using the Danenberg tumor profile (DTP) method. The results were used to examine the correlations between the distributions of the mRNAs and clinicopathological factors, as well as the significance of their expression as a prognostic factor. Patients with poorly differentiated cancers in their tissues showed a significant increase in the mRNA expression of TS and OPRT. The increases in the TP mRNA content were proportional to an increase in the Gleason scores. The prognosis was significantly poorer in those cases with a high expression of TS or OPRT mRNA and a low expression of DPD mRNA. In conclusion, the expression levels of mRNAs for TS, DPD and OPRT among the enzymes related to nucleic acid metabolism are useful as prognostic factors in patients with prostate cancers.