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摘要:
目的 探讨膀胱癌低级别与高级别两条通路在发生发展过程中的分子变化规律。 方法 应用PCR或低变性温度共扩增PCR(COLD-PCR)与Sanger直接测序法检测88例膀胱癌及10例对照组织中fgfr3、p53与h-ras基因突变状况, 以及MRP-1/CD9 mRNA表达水平及各基因与肿瘤复发之间的关系。Logistic回归及相关性分析比较各基因在肿瘤复发中的意义及相互关系。 结果 癌组织中p53突变率随病理分期及分级的增加而增加, MT-p53患者复发率高于WT-p53;fgfr3突变率则与之相反; 低级别病理分期及分级膀胱癌以MT-fgfr3/WT-p53基因型为主, 高级别病理分期及分级膀胱癌以WT-fgfr3/MT-p53基因型为主。h-ras突变率为11.4%(10/88), 主要分布于低级别病理分期及分级膀胱癌中。MRP-1/CD9 mRNA表达随病理分期及分级的增高而降低, 其表达与p53突变率呈负相关, 与fgfr3突变率呈正相关。在膀胱癌患者中WT-fgfr3复发危险为MT-fgfr3的3.88倍, MT-p53复发危险为WT-p53的4.53倍。 结论 低级别病理分期及分级膀胱癌发生发展中以fgfr3及h-ras基因突变为主, 高级别病理分期及分级肿瘤以p53基因突变、MRP-1/CD9 mRNA表达降低为主。fgfr3与p53突变是预测膀胱癌复发的有力指标, 在膀胱癌发生发展中分别代表不同的遗传学通路, 但低级别与高级别两通路有互相重叠。 -
关键词:
- 膀胱癌 /
- 基因突变 /
- 预后 /
- 低变性温度共扩增PCR
Abstract:Objective To investigate the molecular changes in bladder urothelial carcinoma via different pathways. Methods Polymerase-chain reaction (PCR) or coamplification at low denaturation temperature-PCR and Sanger direct sequencing were performed to detect the status of fgfr3, p53, and h-ras gene mutations in 88 tissue samples of human bladder cancer and 10 normal control tissues. The relative mRNA expression levels of motility-related protein-1 (MRP-1)/CD9 and the relationship between genes and tumor recurrence were also determined. Logistic regression and relative analyses were conducted to compare the significance and interrelation of genes among tumor recurrences. Results The mutation rate of p53 increased as pathological grades and stages increased. Recurrence rate was higher in patients with MT-p53 genotype than in patients with WT-p53 genotype. Conversely, the mutation rate of fgfr3 gene decreased as pathological grades and stages increased. Recurrence rate was also higher in patients with WT-fgfr3 genotype than in patients with MT-fgfr3 genotype. In low-grade and early stage tumors, MT-fgfr3/WT-p53 was the most prevalent genotype; in high-grade and late stage tumors, WT-fgfr3/MT-p53 was the most prevalent genotype. The mutations of h-ras were mainly observed in low-grade tumors in early stages. Moreover, the relative mRNA levels of MRP-1/CD9 decreased as pathological grades and stages increased. The mRNA levels of MRP-1/CD9 were negatively correlated with p53 mutations and positively correlated with fgfr3 mutations. Logistic regression analysis results showed that patients with WT-fgfr3 genotypes exhibited 3.88 times higher relative risk of tumor recurrence than those with MT-fgfr3 genotypes; by contrast, patients with MT-p53 genotypes exhibited 4.53 times higher relative risk of tumor recurrence than those with WT-p53 genotypes. Conclusion Fgfr3 and h-ras gene mutations may play important roles in tumorigenesis of low-grade and early stage bladder cancer. p53 gene mutation and mRNA levels of MRP-1/CD9 may be implicated in the tumorigenesis of high-grade tumors in late stage of bladder cancer. In general, the two variants of urothelial carcinoma exhibit distinct genetic defects. fgfr3 gene mutation revealed a pathway of favorable prognosis, and p53 gene mutation demonstrated a pathway associated with poor prognosis. -
表 1 fgfr3、p53、h-ras、MRP-1/CD9及内参基因以mRNA为模板的引物序列
Table 1. Primer sequences using mRNA as templates of fgfr3, p53, h-ras, MRP-1/CD9, and reference genes
表 2 膀胱癌复发的多因素非条件Logistic回归分析
Table 2. Multifactor logistic regression analysis of patients with bladder cancer recurrencce
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