摘要
目的探讨寡核苷酸芯片技术在我国肝细胞癌p53基因点突变发生频率及形式,并以DNA测序法进行验证。方法参照国际p53突变公共数据库资料,以发生频率最高的7个点突变序列设计探针,制备p53基因点突变专用寡核苷酸芯片,应用该技术检测我国肝细胞癌p53基因7个常见突变位点的突变频率及形式,以DNA测序法验证结果,比较不同分组时p53基因点突变差异。结果共检测肝细胞癌石蜡包埋标本54例,p53基因突变率为38.9%(21/54)。54例中广西肝癌高发病区组17例,广西低发病区组(低发区组)19例,区(省)外组18例,三组突变率分别为47%、21%、50%。p53突变主要发生在249编码区,主要突变形式为249ser突变(由AGG→AGT,第三碱基G→T颠换);以DNA测序法对结果进行验证,两种技术检测结果重合率100%。结论寡核苷酸芯片技术可作为检测肝癌p53基因突变的一种新的、可靠和便捷的手段;我国肝癌不同地区之间p53基因突变在频率及形式上既有差异性又有类似性,可能预示不同地区肝细胞癌的病因学异同性。
Objective To study for application of oligonucleotide microarray on detecting the p53 gene point mutation in the paraffin specimens of hepatocellular carcinoma(HCC) from China,compare the point mutation rate of p53 gene with the various groups of HCC.Methods Oligonucleotide microarray which spoted 7 altofrequency mutation sites were designed,according to international data base of p53 gene mutation,and point mutation of 7 codes of p53 gene were detected by the technique of oligonucleotide microarray,and validate the results by the methods of measure sequence.Results 54 specimens of HCC were determined,The total rate of p53 point mutation is 38.9%(21/54),the high,the low incidence of HCC from Guangxi and the non-Guangxi groups were 47%,21%,50%,respectively.The main mutational form was 249ser(AGG→AGT).The results detected with the methods of oligonucleotide chips and sequence were equal respectively.Conclusion The point mutational hotspot of HCC p53 gene was at 249 code.Oligonucleotide chips can be applied to detect gene point mutation rapidly,high efficiently.
出处
《内科》
2006年第1期12-15,99,共5页
Internal Medicine
基金
2004年广西壮族自治区科技厅自然科学基金项目(项目号:桂科自0447026)
2003年广西壮族自治区卫生厅自筹基金项目(项目号:桂卫ZZ2003085)
