Wang Changyu, Weng Yanjie, Wang Hongyan, Shi Ying, Ma Ding
Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2010 Aug;30(4):494-9. doi: 10.1007/s11596-010-0456-y. Epub 2010 Aug 17.
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, TP, 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.
研究了胸苷酸合成酶（TS）、胸苷磷酸化酶（TP）和二氢嘧啶脱氢酶（DPD）的mRNA和蛋白表达及其与预后的关系。采用实时定量RT-PCR（Taqman法）检测106例福尔马林固定石蜡包埋的上皮性卵巢癌组织及29例正常卵巢组织中TS、TP和DPD的mRNA表达。以TATA盒结合蛋白（TBP）作为内参基因。研究TS、TP、DPD表达与临床病理特征之间的关系。采用抗生物素蛋白-生物素-过氧化物酶免疫组织化学法检测同一批患者中TS、TP和DPD的蛋白定位及表达。肿瘤组TS和TP的mRNA表达水平显著高于正常对照组，肿瘤组织中TS和TP的mRNA平均值分别为6.14±0.62和0.59±0.06，正常组织中分别为0.71±0.14和0.16±0.04。肿瘤组DPD的mRNA表达水平（0.11±0.02）显著低于正常对照组（0.38±0.05）。不同病理分级和临床分期的TS和TP mRNA表达水平差异有统计学意义（P<0.05），但组织学亚型与TS和TP mRNA表达无显著相关性。DPD基因表达与任何临床病理参数均无显著相关性。免疫组织化学显示，TP蛋白主要分布于细胞核，TS和DPD主要分布于细胞质。TS、TP和DPD的蛋白表达强度与mRNA表达水平一致。结论：上皮性卵巢癌中TS、TP的mRNA和蛋白表达水平显著升高，DPD的mRNA和蛋白表达水平显著降低。TS和DPD的表达水平与患者的预后和生存相关。TS、TP和DPD的联合基因表达水平代表了预测卵巢癌临床结局的一个新变量。TS、TP和DPD表达水平与生存的相关性提示这些基因在肿瘤发生和进展中具有重要作用。
