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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 detect the mutation of p53 gene in gastric cancer tissues and peritoneal washings by denaturing high performance liquid chromatog-raphy (DHPLC), and to explore the possibility of using this technique as an ideal method for detecting peritoneal subclinical metastasis of gastric cancer. Methods: p53 gene mutation in 45 cases of gastric cancer tissues and peritoneal washings was detected by DHPLC and sequenced. Results: The mutation rate of p53 gene was 20.0% (9/45) in gastric cancer tissues, and 3 of 9 mutations were not reported. The mutation rate of p53 gene in intestinal type gastric cancer was 35.3% (6/17) Diffuse gastric cancer 10.7% (3/28), P <0.01. Two cases of p53 gene mutation were detected in the peritoneal washings, and the detection rate was 22.2% (2/9). The results of sequencing showed that there were 5 (AAG> AGG, Lys 132Arg) and 6 exon (CTG> CCG, Leu188Pro) were the same as those in primary cancerous tissues, all of which were negative in peritoneal wash cytology. Conclusion: DHPLC can be used to detect the mutation of p53 gene in primary foci and peritoneal washings of patients with gastric cancer, and it may be an ideal method for predicting peritoneal subclinical metastasis of gastric cancer if multiple indicators are combined effectively.