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。     

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.     

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。     

晚期胃癌患者血清TS、TP表达水平与氟尿嘧啶为基础的化疗疗效及预后的相关性

背景与目的:胸苷酸合成酶(thymidylate synthetase,TS)和胸苷酸磷酸化酶(thymidinephosphorylase,TP)是氟尿嘧啶类药物在体内代谢的关键酶,与胃癌患者接受氟尿嘧啶药物化疗疗效和预后密切相关。本研究旨在探讨晚期胃癌患者血清中TS、TP表达水平与氟尿嘧啶为基础的化疗疗效及预后的相关性。方法:收集2006年1月—2008年10月于北京大学肿瘤医院消化肿瘤内科治疗的109例一线接受氟尿嘧啶类药物为基础化疗的晚期胃癌患者化疗前的血清标本,运用酶联免疫吸附试验(ELISA)方法检测血清中TS、TP表达水平,分析TS、TP表达水平与化疗疗效及患者预后的相关性。结果:近端胃癌患者的TS表达水平高于远端胃癌患者(P=0.035),腹膜转移患者的TP表达水平明显低于无腹膜转移患者(P<0.001)。TS高表达患者临床获益率(clinical benefit rate,CBR)高于低表达患者(68.75%vs 47.1%,P=0.029),与患者生存无明显相关。TP高表达患者的中位无进展生存期(progression-free survival,PFS)与总生存期(overall survival,OS)均长于低表达患者(PFS:177 d vs 113 d,P=0.018;OS:399 d vs 234 d,P=0.227)。联合分析TS、TP表达发现,TS低表达且TP高表达患者的中位PFS和OS明显长于TS、TP均低表达患者(PFS:182 d vs 87 d,P=0.042;OS:429 d vs 223 d,P=0.019)。Cox多因素生存分析表明,在TS低表达时,TP表达水平是预后的独立影响因素(HR=1.913,95%CI:1.027~3.564,P=0.041)。结论:在接受氟尿嘧啶为基础化疗的晚期胃癌患者中,TS高表达患者的疗效较好,TP高表达患者的生存期较长,特别是TS低表达且TP高表达患者的生存期明显延长。     

结直肠癌患者手术前后胸苷酸磷酸化酶(TP)的表达及临床意义

目的:研究结直肠癌组织和血清中胸苷磷酸化酶(thymidine phosphorylase TP)表达与病理及生物特性之间的关系。方法:对50例结直肠癌标本、20例正常结直肠黏膜标本及18例结直肠良性病变组织进行免疫组化S-P法检测,测定TP表达情况;ELISA法测定结直肠癌患者术前、术后血清TP水平及20例健康志愿者血清TP水平。结果:结直肠癌组织中胸苷磷酸化酶表达的阳性率显著高于正常结直肠黏膜、结直肠良性病变(P<0.05);有淋巴结转移的结直肠癌组TP表达阳性率高于无淋巴结转移组(P<0.05);结直肠癌组血清TP水平较健康组为高,差别具有统计学意义(P<0.05);DukesA-B期和DukesC-D期患者之间血清TP水平比较差别具有统计学意义(P<0.05)。术前和术后结直肠癌患者血清TP水平差别具有统计学意义(P<0.05)。结论:结直肠癌Dukes分期较晚及有淋巴结转移者,其瘤组织中TP表达的阳性率高;血清胸苷磷酸化酶的水平与患者的肿瘤病理分期有关,分期越晚表达水平越高;胸苷磷酸化酶在组织中的表达与其血清的水平呈正相关性,血清胸苷磷酸化酶的水平可以反应其组织中的表达水平。     

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.     

结直肠癌患者胸苷酸磷酸化酶活性与XELOX新辅助化疗疗效的关系

目的 研究结直肠癌中胸苷酸磷酸化酶(TP)活性与XELOX新辅助化疗疗效之间的关系.方法 收集64例采用XELOX方案化疗的结直肠癌患者临床资料,根据TP酶活性不同分为TP高酶活性组(32例)和TP低酶活性组(32例),统计两组患者的化疗效果以及术后5年生存期.结果 TP高酶活性组患者化疗后病灶减小程度高于TP低酶活性组,周围组织的浸润程度低于TP低酶活性组,同时患者的术后生存率亦高于TP低酶活性组,差异均具有统计学意义(P＜ 0.05).结论 结直肠癌中胸苷酸磷酸化酶活性越高,XELOX化疗敏感性越高,化疗效果越好,预后相对优于TP酶活性低者.     

结直肠癌组织胸苷磷酸化酶表达临床意义研究的现状

目的:总结近年来关于胸苷磷酸化酶(TP)在结直肠癌组织中表达临床意义的研究现状.方法:应用PubMed、Elsevier Sciencedirect、CNKI及万方全文数据库检索系统,以胸苷磷酸化酶和结直肠癌为关键词,检索2005-01-2011-06发表的相关文献.纳入标准:1)TP在结直肠癌及转移组织中的表达及其促血管新生、抗细胞凋亡等生物学作用.2)TP与5-氟尿嘧啶(5-FU)前体类药物靶向治疗的关系.根据纳入标准分析文献29篇.结果:TP对于结直肠癌的影响是双重性的,其在肿瘤血管新生、转移、复发以及抗细胞凋亡等过程中均发挥重要作用,高表达TP的患者往往预后不良.另外,TP又在5-FU前体药物的化疗过程中起着关键作用,增加TP的表达很可能可以增强5-FU前体药物的化疗效果.结论:TP在结直肠癌组织中的表达对结直肠癌靶向治疗和预后有着非常重要的临床意义.     

胸苷酸合成酶基因3′-UTR多态性与晚期胃癌化疗敏感性关系的研究

目的:研究胸苷酸合成酶(TS)基因3′-UTR多态性与胃癌对5-FU化疗敏感性的关系.方法:收集经病理学确诊的晚期胃癌106例,所有病例化疗前抽静脉血,提取白细胞DNA,用PCR-RFLP技术检测TS3′-UTR基因型.所有患者均经5-FU为基础的化疗方案治疗.以WHO实体瘤疗效评定标准和毒性评定标准评价疗效和毒性.结果:(1)106例胃癌患者中TS+6/+6bp、+6/-6bp、-6/-6bp基因型频度分别为7.6%、44.3%和48.1%,化疗的总有效率为35.9%.(2)TS+6/+6bp、+6/-6bp、-6/-6bp基因型组化疗的有效率分别为0%(0/8)、40.4%(19/47)和37.3%(19/51).TS-6/-6bp基因型组和+6/-6bp组化疗的有效率均显著高于+6/+6bp组(Fisher双侧精确检验,P值分别为0.0452和0.0404).TS-6/-6bp基因型组Ⅱ度以上毒副反应的发生率高于+6bp/+6bp基因型组,但其差异无统计学意义.结论:TS基因3′-UTR多态性与晚期胃癌对5-FU为基础的化疗敏感性相关.TS基因型检测有助于指导晚期胃癌的化疗.     

5-FU耐药免疫组化技术与MTT法敏感性比较

目的:探讨结肠癌对5-氟脲嘧啶(5-FU)产生耐药的机理,从细胞培养水平进一步论证胸苷酸合成酶(TS)对5-FU化疗产生耐药的影响,揭示不同TS蛋白表达水平的结肠癌患者对5-FU治疗的敏感性.方法:应用MTT法对体外培养的30例结肠癌细胞进行5-FU药敏试验,再应用SP免疫组化法对30例结肠癌进行TS检测,然后将两个试验结果进行比较;另采用SP法检测60例结肠癌组织切片及其正常对照组切片中TS蛋白的表达情况.结果:TS高表达组体外培养的结肠癌细胞对5-FU的敏感性明显低于TS低表达组.正常结肠粘膜组织中的TS表达水平高于结肠癌组织,结肠癌组织中TS表达水平与5-FU化疗患者的预后呈负相关,与患者的性别、有无淋巴结转移及癌组织的分化程度无关.结论:TS蛋白表达程度的高低对临床结肠癌患者化疗药物的选择有一定的指导意义:结肠癌组织中TS蛋白的表达水平可作为患者对5-FU为主的化疗疗效及预后的判断指标.     

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.     

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.     

胃癌c-Jun 和MMP-9 表达与其生物学行为的关系*

目的:研究转录因子c-Jun 及下游蛋白MMP-9 在胃癌组织中的表达状况,分析其与临床病理特征和生存之间的关系。方法:应用免疫组化SP法,采用组织芯片技术,分析189 例胃癌组织、54例癌旁组织、41例淋巴结转移灶及32例正常对照组织中c-Jun 和MMP-9 蛋白表达。结果:胃癌中c-Jun 和MMP-9 蛋白的阳性表达率分别为73.0% 和78.3% 。c-Jun 蛋白的表达与分化程度有关（P<0.05）;MMP-9 的表达与浸润深度、淋巴结转移、分化程度及Lauren 分型有关（P<0.05）。 胃癌中c-Jun 和MMP-9 蛋白表达水平呈正相关（P<0.05）。 41例淋巴结转移灶和相对应的41例胃癌组织各因子表达比较发现,MMP-9 阳性表达率有显著性差异（P<0.05）,c-Jun 阳性表达率无显著性差异（P>0.05）。 单因素生存分析显示c-Jun 和MMP-9 阳性组对胃癌患者生存期的影响有统计学意义（P<0.05）,COX
多元回归分析显示c-Jun 和MMP-9 的异常表达均不可作为影响胃癌预后的独立因素（P>0.05）。 结论:c-Jun 和MMP-9 在胃癌组织中均有高表达。c-Jun 阳性表达与胃癌的组织学分级密切相关,提示其异常表达可能是胃癌发生过程中的一个早期分子事件。MMP-9 阳性表达与肿瘤的浸润、转移密切相关,可作为判断胃癌恶性行为的重要生物学标记物。c-Jun 和MMP-9 蛋白表达水平呈正相关。c-Jun 和MMP-9 蛋白阳性表达对于胃癌患者是一个不利的预后因素,尚不能作为一个独立的预后指标。      

lnc-KCNQ1OT1在胃癌组织和细胞中的表达及其临床意义

目的:探讨长链非编码RNA-KCNQ1OT1(lnc-KCNQ1OT1)在胃癌组织和细胞中的表达及临床意义。方法:收集 2010年1月1日至2011年12月31日期间在哈尔滨医科大学附属肿瘤医院行手术治疗的163 例胃癌患者的临床资料,选取组织和细胞通过RT-PCR法检测lnc-KCNQ1OT1的表达情况,根据其表达情况及临床病例数据分析lnc-KCNQ1OT1在胃癌组织和细胞中的临床意义。结果:与癌旁组织比较,lnc-KCNQ1OT1在胃癌组织中的表达水平明显增高（P<0.05）。在胃癌细胞株HGC-27、MGC-803中,lnc-KCNQ1OT1表达水平较胃永生化上皮细胞株GES-1明显升高（P<0.05）。单因素分析显示lnc-KCNQ1OT1的表达与患者年龄、性别、肿瘤大小无关（P>0.05）,与淋巴结转移（P<0.001）明显相关。K-M生存分析显示胃癌患者中lnc-KCNQ1OT1高表达者预后不良（P<0.05）。结论:在胃癌组织和细胞中lnc-KCNQ1OT1的表达量高,其高表达与胃癌患者淋巴结转移相关,并可导致胃癌患者预后不良。lnc-KCNQ1OT1可作为潜在的胃癌预后预测的分子标志物。     

miR-3666在胃癌中的表达及其临床意义

目的:检测胃癌组织中miR-3666的表达并分析其与临床病理特征的关系。方法:采用茎环逆转录实时定量PCR技术检测80例胃癌手术标本与相对应的癌旁组织中miR-3666的相对表达量,并分析其与胃癌临床病理参数和预后的关系。结果:miR-3666在胃癌组织中表达与癌旁组织相比显著下调（P<0.001）,miR-3666的表达与胃癌淋巴结转移（P=0.002）和TNM分期（P=0.017）相关,与年龄、性别、浸润深度、肿瘤位置及分化程度无关（P>0.05）。合并淋巴结转移的胃癌组织中miR-3666的表达低于无淋巴结转移的胃癌组织（P<0.001）。miR-3666低表达与高表达的患者1年及3年生存率分别为78.0%、22.6%和96.9%、48.1%,中位生存率分别为16个月和29个月,差异有统计学意义（P<0.001）。COX预后分析发现miR-3666低表达和肿瘤TNM分期是胃癌患者预后的独立影响因素。结论:miR-3666在胃癌组织中的表达量明显降低,提示miR-3666可能参与了胃癌的发生发展,并且与胃癌的淋巴结转移相关;miR-3666低表达提示胃癌患者预后不良。     

Thymidylate synthase expression in colorectal cancer: a prognostic and predictive marker of benefit from adjuvant fluorouracil-based chemotherapy.

PURPOSE: We studied the prognostic value of thymidylate synthase (TS) expression in primary colorectal cancer (CRC) and the role of TS expression as a predictor of chemotherapeutic benefit in patients treated with adjuvant chemotherapy. PATIENTS AND METHODS: TS expression was immunohistochemically assessed on tumor sections from 862 patients with CRC Dukes' stages B and C enrolled onto randomized trials evaluating fluorouracil (5-FU)-based adjuvant chemotherapy. RESULTS: TS expression was an independent prognostic factor for disease-free (P =.05) and overall survival (P =.05). In the subgroup treated with surgery alone, TS was an independent prognostic factor for disease-free (P <.001) and overall survival (P =.001), whereas this was not the case in the subgroup of adjuvantly treated patients. Patients whose tumors expressed high TS levels had a tendency to improved outcome after adjuvant therapy (not significant). The group whose tumors expressed the highest TS grade, grade 3 (34% of the patients), had a significantly longer disease-free survival if they were treated with adjuvant therapy compared with surgery alone (multivariate analyses, P =.02), whereas patients whose tumors expressed low TS levels (28% of the patients) had an impaired outcome after adjuvant therapy (multivariate analyses: disease-free survival, P =.01; overall survival, P =.01). CONCLUSION: TS expression predicts for survival independent of Dukes' stage in patients with CRC treated with surgery alone. The study indicates that patients with high TS levels may benefit from adjuvant 5-FU-based chemotherapy. However, patients with low TS levels seem to have a worse outcome when treated with adjuvant chemotherapy.     

MMP-9 在胃癌及其淋巴结组织中表达的临床意义*

目的:研究MMP-9 在胃癌组织中的表达状况,分析其与临床病理特征和生存之间的关系。方法:应用免疫组化SP法,采用组织芯片技术,分析189 例胃癌、54例癌旁组织、41例淋巴结转移灶及32例正常对照组织中c-Jun 和MMP-9 蛋白表达。结果:胃癌中MMP-9 蛋白的阳性表达率为78.3% 。MMP-9 的表达与浸润深度、淋巴结转移、分化程度及Lauren 分型有关（P<0.05）。 41例淋巴结转移灶和相对应的41例胃癌组织各因子表达比较发现,MMP-9 阳性表达率具有显著性差异（P<0.05）。 与MVD进行比较发现,MMP-9 阳性患者的MVD值高于阴性组,差异有统计学意义（P<0.05）。 单因素生存分析显示MMP-9 阳性组对胃癌患者生存期的影响有统计学意义（P<0.05）,Cox 多因素回归分析显示MMP-9 的异常表达不能作为影响胃癌预后的独立因素（P>0.05）。 结论:MMP-9 在胃癌组织中均有高表达。MMP-9 阳性表达与肿瘤的浸润、转移密切相关,可作为判断胃癌恶性行为的重要生物学标记物。MMP-9 在促进血管生成中起重要作用。MMP-9 蛋白阳性表达对胃癌患者来说是一个不利的预后因素,但尚不能作为一个独立的预后指标。