应用聚合酶链反应-单链构象多态性分析检测散发性乳腺癌EMSY基因的突变

Mutations detection of EMSY gene in sporadic breast cancer with PCR - SSCP

目的了解EMSY基因在散发性乳腺癌组织中的突变情况，探讨EMSY基因突变与散发性乳腺癌的关系。方法收集72例散发性乳腺癌组织及18例乳腺良性肿瘤患者的肿瘤组织，常规酚-氯法抽提组织DNA，并应用聚合酶链反应-单链构象多态性分析（PCR—SSCP）的方法，扩增其EMSY基因的20个外显子，然后根据构象改变来分析此基因突变情况。结果72例散发性乳腺癌组织以及18例乳腺良性肿瘤组织中，6例样本存在第9外显子SSCP条带的异常改变，测序结果为第9外显子区GCA（丙氨酸）中的A杂合性缺失，突变率为8．3％（6／72）。结论本课题研究提示广西散发性乳腺癌中存在着EMSY基因第9外显子的缺失突变，并且在乳腺癌患者中突变率高于乳腺良性肿瘤患者。

Objective To get the message of mutatian information of EMSY gene in sporadic breast carcinoma tissues, then investigate the association of EMSY gene mutation and sporadic breast carcinoma disease. Methods Tissue from 72 sporadic breast carcinoma patients and 18 optimum breast tumor patients, being extracted DNA with phenol and chloroform, polymerase chain reaction -single strand con- formation polymorphism （ PCR - SSCP） was used toamplify 20 exons of EMSY and analyze the conformation polymorphism. Results Among the 72 cases of sporadic breast carcinoma specimens and 18 cases of optimum breast tumor specimens, there were 6 breast carcinoma specimens, which the exon 9 have conformation change in single stranded. The sequencing result was confirmed loss of heterozygosity （LOH） and the deletion was A of GCA. The mutation rate was 8.3%. Conclusions The study of this subject shows that it has EMSY gene mutation in Guangxi sporadic breast cancer, and the rate is higher than optimum breast tumor.