摘要
目的 建立纳米捕获探针体系检测血浆中鼠类肉瘤病毒癌基因（K-ras）突变的方法,并探讨K-ras基因单核苷酸多态性突变与胰腺癌临床病理特征及其预后的关系.方法 收集胰腺癌患者72例,所有患者均明确诊断.抽提各血浆标本DNA,应用特异性纳米捕获探针检测K-ras基因12、13位点密码子突变,结合临床资料,分析K-ras基因点突变与胰腺癌临床病理特征的关系及其对预后的影响.结果 72例胰腺癌患者血浆DNA浓度为20-200ng/μl,33例检测到K-ras基因突变,循环阈值（Ct）为21.71-35.61,其中12位点30例,13位点3例,K-ras基因突变率为45.8%（33/72）;其中37例相对早期胰腺癌（Ⅰ＋Ⅱ期）的K-ras基因突变10例,突变率为27.0%（10/37）,而35例中晚期胰腺癌（Ⅲ＋Ⅳ期）的K-ras基因突变23例,突变率为65.7%（23/35）,两者比较差异有统计学意义（x2=10.843,P=0.001）,提示K-ras基因点突变与肿瘤分期相关;72例胰腺癌患者均获得随访,随访时间为2个月至26个月,平均随访时间9.7个月,通过随访分析K-ras基因突变者1年生存率为42.6%,K-ras基因野生型患者1年生存率为73.4%,差异有统计学意义（x2=5.335,P=0.017）.结论 采用纳米捕获探针体系能够快速痕量检测血浆中K-ras基因突变,其与胰腺癌的分期、预后密切相关.
Objective To establish a method of detection kirsten rat sarcoma viral oncogene mutations in plasma by a nano capture probe system, and to explore the relationship between clinical pathology stage, or prognostic and K-ras single nucleotide polymorphism mutations in patients with pancreatic cancer.Methods The plasma samples of 72 patients with pancreatic cancer were collected from June 2013 to September 2015.The diagnosis of all patients were explicit.The DNA were extracted from all plasma samples and were detected for the codon 12 and 13 mutation of K-ras gene by nano capture probe.The relationship between clinical pathology factors, or prognostic and K-ras point mutations in pancreatic cancer was analyzed via clinical data.Results The plasma DNA concentration of 72 patients with pancreatic cancer was 20-200 ng/μl, 33 patients were detected for K-ras gene mutation, the cycle threshold （Ct） value was 21.71-35.61, including 30 patients for codon 12 mutation and 3 patients for codon 13 mutation, the K-ras mutation rate was 45.8% （33/72）.10 of 37 patients with early pancreatic cancer （stage Ⅰ and Ⅱ） were detected for K-ras gene mutation, the mutation rate was 27.0% （10/37）, but 23 of 35 patients with advanced pancreatic cancer （stage Ⅲ and Ⅳ） were detected for K-ras gene mutation, the mutation rate was 65.7% （23/35）, the difference is statistically significant （x2=10.843, P=0.001）, it is suggested that K-ras mutation in plasma is ralated to tumor stage.All 72 patients with pancreatic cancer were followed up, the follow-up time was 2 months to 26 months, the average follow-up time was 9.7 months.The one-year survival rate of patients with K-ras mutation is 42.6%, and the one-year survival rate of patients with wild genotype is 73.4%, the difference is statistically significant （x2=5.335, P=0.017））.Conclusion The nano capture probe system could certainly detect K-ras mutation in rare plasma DNA.K-ras mutation in plasma is related to TMN stage and prognosis of pancreatic cancer, and K-ras mutation in plasma DNA is a predictive biomarker for a poor prognosis of pancreatic cancer patients.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2017年第3期375-377,共3页
Chinese Journal of Experimental Surgery
基金
浙江省科技厅公益技术研究社会发展项目（2014C33139）
关键词
胰腺癌
预后
突变
磁性纳米粒子
Pancreatic cancer
Prognosis
Mutation
Magnetic nanoparticles
