摘要
目的检测涎腺多形性腺瘤基因突变方式及野生型 p53对突变基因的修复。方法留取4例涎腺多形性腺瘤新鲜标本,进行原代细胞培养。采用野生型 p53基因转染培养细胞,通过逆转录聚合酶链反应检测转染后的基因表达。提取转染后各例细胞 DNA 及对应的肿瘤组织 DNA,分别进行 PCR 反应扩增,聚丙烯酰胺凝胶电泳及核酸序列测定分析。结果 PCR-单链构象多态性分析屁示,3例出现异常电泳带。核酸序列测定结果显示,第6外显子的第203号密码子点突变;第8外显子的第272至290号密码子之间出现碱基缺失和碱基插入。野生型 p53转染后的细胞 DNA 序列分析显示,p53第6和第8外显子的5个突变位点均恢复正常。结论涎腺多形性腺瘤发生过程中具有较高频率的 p53基因突变,突变方式多样,涉及多个密码子;外源性野生型 p53可有效地修复涎腺多形性腺瘤细胞中突变的 p53基因位点。
Objective To analyze the original mutated codon of p53 gene of salivary plcomorphic adenoma （SPA） and to evaluate the repair effects of wt-p53 on SPA cells. Methods Four cases of SPA were obtained from clinical fresh samples and SPA cells were separated and cultured, and then the cells were transduced by Ad-wt-p53. The cells and the corresponding tumor tissue DNA were extracted, PCR and single strand conformational polymorphism （SSCP） and DNA sequencing analysis were performed. Results PCR-SSCP analysis showed 3 out of 4 SPA with abnormal exon 8 and abnormal exon 6. DNA sequencing analysis showed that exon 6 point mutation was found at codon 203 （ GTG→GCG）, poly-codon mutations were found in exon 8 at codon 272 （GTG→GT□） ,275 （TGT→T□T） ,276 （GCC→□CC） and at codon 290 （CGC→CGCC）. After transduced by Ad-wt-p53, all of the mutated codons were repaired. Conclusions p53 gene mutation involved many codons that occurred frequently in the tumorigenesis of SPA. Exogenous wt-p53 could compensate and repair all the mutated p53 codons of SPA cells. SPA cells transduced by Ad-wt-p53 showed the typical apoptosis.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2007年第3期144-147,共4页
Chinese Journal of Stomatology
基金
国家自然科学基金(30271422)
河北省自然科学基金(C2004000624)
关键词
涎腺
腺瘤
多形性
基因
P53
Salivary gland
Adenoma,pleomorphic
C.ene,p53
