摘要
目的了解大肠癌组织及粪便中P53蛋白K-ras及APC基因的突变状况,了解粪便脱落细胞基因检测的可行性及其临床意义方法采用S-P法对27例大肠癌患者的癌组织及其粪便脱落细胞 P53蛋白进行检测,以22例大肠癌组织及10倒粪便和9例正常大肠粘膜组织为研究对象,采用PCR-RFLP银染方法检测K-ras基因12,13密码子及应用PCR-SSCP银染技术检测APC基因突变集中区(MCR区)的突变状况结果粪便脱落细胞P53表达阳性率为37％(10/27),与相应患者大肠癌组织P53检测一致率为85％(23/27)、K-ras12密码子突变检出敏感性癌组织为73％(16/22),粪便为50％(5/10).粪便与相应患者的癌组织检测符合率为90％(9/10);癌组织及粪便中均未检出K-ras13密码子突变.癌组织中APC基因突变率为41％(9/22),粪便中APC突变体阳性检出率为40％(4/10),癌组织与相应患者的粪便APC基因检测一致率为90％(9/10).P53,K-ras及APC三个基因联合检测癌组织敏感性为100％,粪便为90％.结论 P53,K-ras及APC基因突变为散发性大肠癌常见的分子事件,联合基因检测可提高对大肠癌检测的敏感性;粪便中基因突变体检测结果忠实反映了癌组织突变状况,对其检测有望成为大肠癌诊断及筛查的无创分子途径.
AIM To investigate the mutation of P 53, K-ras and APC gene in human stools and its significance in diagnosis of colorectal cancer. METHODS Tumor tissue and colorectal exfoliative cells in stools were collected from 27 patients and then, using S-P method, P 53 protein was detected. The mutation of K- ras code 12, 13 and APC gene at exon 15 MCR in 22 specimens of tumor mucosa, 9 specimens of normal mucosa and 10 samples of stool were determined by polymerase chain reaction amplification and restriction enzyme analysis (PCR-RFLP) and polymerase chain reaction and its single strand conformation polymorphism (PCR-SSCP) respectively. RESULTS In ten (37%) of 27 cases of colorectal cancer, P 53 was positive in the exfoliative cells. The agreement rate of P 53 expression in tumor and exfoliative cells was 85% (23/27). In sixteen (73%) of 22 samples of tumor tissue and 5 (50%) of 10 samples of stool (511%), the mutations of K-ras code 12 were positive. The agreement rate of K-ras code 12 mutations in tumor and stool was 90% (9/10). No K-ras code 13 mutations were observed in stool and tumor mucosa. The mutations were detected in nine (41%) of 22 specimens of tumor mucosa and 4 (40%) of 10 samples of stool. The agreement rate in tumor and stool was 90% (9/10). Detection of P 53, K-ras and APC gene mutation at same time, the sensitivity for colorectal cancer in tissues vs stool was 100% vs 90%. CONCLUSION The mutation of P 53, K-ras code 12 and APC frequently occurs on colorectal carcinoma, detection of P 53, K-ras and APC gene mutation could raise the positive rate for colorectal cancer. Investigation of gene mutation in the stool reflects the real situation of that in the tissue, its detection may be a useful noninvasive molecular approach for screening and diagnosis of colorectal carcinoma.
出处
《世界华人消化杂志》
CAS
2001年第7期771-775,共5页
World Chinese Journal of Digestology
关键词
病理学
诊断
粪便
细胞学
蛋白质P53
大肠癌
colorectal neoplasms/pathology
feces/cytology
diagnosis
protein p53/analysis
gene, ras
gene, APC
