胃癌组织和腹腔冲洗液p53基因突变的DHPLC技术检测  

Detection of p53 gene mutation in cancer tissues and peritoneal lavage of gastric cancer patients by denaturing high-performance liquid chromatography

作　　者：鲁翀[1] 徐岩[1] 任群[2] 王振宁[1] 敖阳[2] 姜莉[2] 罗阳[2] 张学[2] 徐惠绵[1] 

机构地区：[1]中国医科大学附属第一医院肿瘤外科/普外教研室胃肠肿瘤外科,辽宁沈阳110001 [2]中国医科大学医学基因组学研究室,辽宁沈阳110001

出　　处：《中华肿瘤防治杂志》2009年第8期613-616,共4页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:应用部分变性高效液相色谱(denaturing high performance liquid chromatog-raphy,DHPLC)检测胃癌组织、腹腔冲洗液中p53基因突变,并探讨该技术成为检测胃癌腹膜亚临床转移理想方法的可能性。方法:应用DHPLC对45例胃癌组织及腹腔冲洗液中p53基因突变进行检测,并测序验证。结果:胃癌组织p53基因的突变率为20.0%(9/45),9例突变中有3例突变均未见报道;p53基因在肠型胃癌中突变率35.3%(6/17)明显高于弥漫型胃癌10.7%(3/28),P<0.01。腹腔冲洗液中检出p53基因突变2例,检出率为22.2%(2/9),经测序证实分别位于第5外显子(AAG>AGG,Lys132Arg)和第6外显子(CTG>CCG,Leu188Pro),均与原发癌组织中相同,这2例患者腹腔冲洗液细胞学检测均为阴性。结论:DHPLC可应用于胃癌患者原发癌灶及腹腔冲洗液中p53基因突变的检测,而且如有效地联合检测多个指标,则可能成为预测胃癌腹膜亚临床转移的理想方法。OBJECTIVE： To investigate p53 gene mutation in cancer tissues and peritoneal lavage of gastric cancer patients by denaturing high-performance liquid chromatography （DHPLC）. METHODS： The mutation of p53 gene was detected by DHPLC in the specimens of cancer tissues and peritoneal lavage of 45 patients with gastric cancer, then sequencing was performed. RESULTS： The mutation rate of p53 gene in gastric cancer tissues was 20.0% （9/45） ; the mutation rate in in testinal type gastric cancer （35.3%, 6/17） was higher than that in diffuse-type gastric cancer （10.7%, 3/28）,P〈0.01. Two cases of p53 mutation （Lys132Arg at exon 5 and Leu188Pro at exon 6）were found in peritoneal lavage and were just identical to the mutation found in the primary lesions, respectively. While no free cancer cell was found in the peritoneal lavage cytology of these 2 patients. CONCLUSIONS： DHPLC is a convenient method for detecting the mutation in cancer tissues and peritoneal lavage of gastric cancer patients. DHPLC may become a kind of ideal method for prediction of subclinical peritoneal metastasis if more appropriate DNA variations are detected together.

关 键 词：胃肿瘤 腹腔灌洗 基因 p53 色谱法 

分 类 号：R735.2[医药卫生—肿瘤]