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背景:肿瘤相关基因在癌瘤发生、发展、转移及预后过程中的作用如何一直是医学界研究的难点。目的:了解变性梯度凝胶电泳(DGGE)和自动DNA序列分析方法检测肿瘤基因变异及p53基因、p53蛋白在大肠癌发生、转移过程中的动态变化,为大肠癌预后评估提供依据。设计:以组织标本为研究对象的单一样本研究。单位:一所军医大学医院肿瘤科。对象:收集1994-01/2000-12解放军第一军医大学南方医院住院治疗的41例因大肠癌接受大肠根治性切除并于手术后5个月至5年内因肝转移接受肝切除术患者的大肠癌原发灶及肝转移灶标本。方法:以DGGE及自动DNA序列分析法检测41例大肠癌原发病灶和肝转移灶p53外显子5～11的基因突变。以免疫组织化学方法检测p53蛋白表达。主要观察指标:①p53基因突变在DGGE中的检出情况及突变。②p53基因的序列分析。③p53免疫组织化学染色结果。结果:41例中24例有p53基因突变(62%),其中6例仅在肝转移灶发现p53基因突变,其余均为原发灶、转移灶有一致性的突变。另有3例原发灶即有p53基因突变的患者,在转移灶除保留原有突变外,还出现新增加的突变。在原发、转移灶同时有突变的16例中,14例呈现突变的p53碱基峰和正常峰之比在肝转移灶明显高于大肠癌原发灶(P<0.001)。p53免疫组织化学染色结果和DGGE,DNA序?
Background: How the role of tumor-related genes in the carcinogenesis, development, metastasis and prognosis of cancer has always been a challenge in medical research. OBJECTIVE: To understand the variation of tumor gene and the dynamic changes of p53 and p53 proteins in the carcinogenesis and metastasis of colorectal cancer by denaturing gradient gel electrophoresis (DGGE) and automated DNA sequence analysis, and to provide evidence for the prognosis evaluation of colorectal cancer. Design: A single-sample study of tissue specimens. Unit: a Military Medical University Hospital oncology. PARTICIPANTS: Forty-one patients who underwent radical resection of the large intestine by colorectal cancer who underwent inpatient treatment at the Southern Hospital of the First Military Medical University from January 1994 to December 2000 were enrolled, and the large intestine underwent liver resection within 5 months to 5 years after surgery Primary cancer and liver metastases specimens. Methods: The gene mutations of p53 exon 5 ~ 11 in 41 cases of colorectal carcinoma primary and hepatic metastases were detected by DGGE and automated DNA sequence analysis. P53 protein expression was detected by immunohistochemistry. MAIN OUTCOME MEASURES: ① Detection and mutation of p53 gene mutation in DGGE. ② p53 gene sequence analysis. ③ p53 immunohistochemical staining results. Results: Twenty-four of 41 cases had p53 gene mutation (62%). Of the 6 cases, only p53 gene mutation was found in liver metastasis and the others were primary and metastatic. Another 3 cases of primary tumor p53 gene mutation in patients with metastasis, in addition to retain the original mutation, there are also new mutations. Among the 16 cases with primary and metastatic mutations, the ratio of p53 base peak to normal peak in 14 cases was significantly higher than that in primary liver cancer (P <0.001). p53 immunohistochemical staining results and DGGE, DNA sequence?