胃肠道间质瘤c-kit基因全部外显子的突变研究

目的 c-kit基因全部外显子在胃肠道间质瘤疾病中的突变情况,并探讨其意义.方法 收集2010年9月至2012年12月胃肠道间质瘤手术切除标本62例,其中胃间质瘤41例,小肠14例,直肠4例,结肠1例,腹膜2例,年龄20～82岁,男性40例,女性22例,中位年龄54岁,术前筛查均无GIST家族史,患者术前均未接受过放疗或化疗,都是经术中病理报告和最终石蜡切片病理报告确诊的GIST标本.设计特异性PCR引物,用肿瘤组织提取的DNA为模板进行PCR扩增后测序,研究c-kit基因突变在胃肠道间质瘤组织中的突变规律.结果 在62例胃肠道间质瘤组织标本中有53例c-kit基因发生突变,突变率85.5%（53例/62例）,PCR产物突变率为3.8%（62例/1612例）,外显子2突变6例（9.7%）,外显子8突变7例（11.3%）,外显子10突变14例（22.6%）,外显子11突变8例（12.9%）,外显子21突变27例（43.5%）.此外第21号外显子有8例位于c-kit基因第80778位点的C突变成T,有19例位于c-kit基因第80846位点的G突变成A.结论 胃肠道间质瘤中c-kit基因突变率高达85.5%,表明c-kit基因突变可能对胃肠道间质瘤的发生发展起重要作用.

Study of the mutation for all exons of the c-kit gene in human gastrointestinal stromal tumor

Objective To explore the mutation of all exons of the c-kit gene in gastrointestinal stromal tumor disease and its clinical significance. Methods 62 cases of gastrointestinal stromal tumor operation resection specimens were collected from the First Affiliated Hospital of Guangxi Medical U- niversity from September 2010 to December 2012, Samples were stored at - 20℃ freezer. The DNA was extracted using phenol-chloroform method to detect the content and purity of DNA with ultravio- let spectrometer. Software was used to to design specific PCR primers in accordance with the primer design requirements. DNA of tumor tissue was extracted, followed by PCR amplification. To detect PCR products by agarose gel electrophoresis, PCR products were sequenced（Sequenced by Shanghai SANGON company） using spss 13. 0 statistical software package for statistical, aiming to study the law of mutations of c-kit gene in gastrointestinal stromal tumor tissues. Results There were 53 cases of c kit gene mutation in 62 cases of gastrointestinal stromat tumors, The mutation rate was 85. 5% （53 cases/62 cases）,PCR product of the mutation rate was 3.8% （62 PCR products/1612 PCR prod- ucts）. The mutation of exon2 in 6 cases （9. 7%）, the mutation of exon 8 in 7 cases （11.3%）, the mutation of exon 10 in 14 cases （22. 6%）, the mutation of exon 11 in 8 cases （12. 90%）, the mutation of exon 2l in 27 cases （43.5%）. Conclusion The rate of mutation in the c-kit gene was as high as 85.5% in gastrointestinal stromal tumor, the results showed that the mutation of c-kit gene may play an important role in the occurrence and development role of gastrointestinal stromal tumor, and pro- vides experimental basis and new ideas for further study of gastrointestinal stromal tumors. Key words] Gastrointestinal stromal tumor % e-kit gene; Mutation; Sequencing