摘要
目的 比较分析人骨肉瘤组织mtDNA控制区的序列和正常人的差异。方法 用ApaⅠ ,AvaⅡ ,BamHⅠ ,EcoRⅤ ,HaeⅢ和KpnⅠ共 6种限制性内切酶 ,对 2 0例骨肉瘤组织进行了PCR产物限制性片段长度多态性 (PCR RFLP)分析。结果 2 0例骨肉瘤组织DNA均扩增出长 14 5 1bp的线粒体控制区特异性目的片段 ,没有发现长度多态性。 8例骨肉瘤组织在mtDNA 165 19位核苷酸处发生了T→C的突变 ,占 40 % (8/ 2 0 ) ,其中骨母细胞型 4例 ,占 40 % (4 / 10 ) ,软骨母细胞型 2例 ,占 40 % (2 / 5 ) ,纤维母细胞型 2例 ,占 40 % (2 / 5 ) ,各亚型之间没有显著性差异 (P >0 0 5 ) ;7例组织第 1613 2位核苷酸处的A缺失 ,占 3 5 % (7/ 2 0 ) ,其中骨母细胞型 3例 ,占 3 0 % (3 / 10 ) ,软骨母细胞型 2例 ,占 40 % (2 / 5 ) ,纤维母细胞型 2例 ,占 40 % (2 / 5 ) ,各亚型之间没有显著性差异 (P >0 0 5 ) ;4例组织既有 165 19位核苷酸处T→C的突变 ,又有 1613 2位A的缺失 ,占 2 0 % (4 / 2 0 )。这两种突变在 2 0例骨肉瘤病例以及各亚型中占的比例均显著高于正常人群相应位点的多态性变异。结论 这两种突变在骨肉瘤细胞mtDNA中发生率较高 ,可能在骨肉瘤的发生发展中发挥重要的作用。
Objective To determine if the mutation rates of mitochondrial DNA (mtDNA) control region in patients with osteosarcoma are higher than those of the healthy human, and to discover the possible specific mutations of mtDNA control region in patients with osteosarcoma. Methods Total DNA, extracted from formalin-fixed and paraffin-embedded tissues from 20 cases of osteosarcoma by Goelz's method, was studied by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) for mutations within the mtDNA control region. Results The T16519C mutation was detected in 8 out of 20 (40%) samples and the rates of this mutation were 40% (4/10), 40% (2/5), and 40% (2/5) in osteoblastic, fibroblastic, and chondroblastic osteosarcoma, respectively. There was no significance between three subtypes of osteosarcoma (P>0.05). The A deletion in the nucleotide position 16132 (5′ end) was found in 7 (35%) out of 20 samples. No significant difference was (P>0.05) found between the three subtypes of osteosarcoma with rates of the A deletion being 30% (3/10), 40% (2/5), and 40% (2/5) in three subtypes, respectively. The T16519C mutation and the A deletion in the nucleotide position 16 132 (5′ end) were co-detected in 4 (20%) out of 20 samples. The levels of the two mutations in patients with osteosarcoma were significantly higher than those in healthy human. Conclusion The two mutations have high rates in osteosarcoma and may play an important role in carcinogenesis and progression of osteosarcoma.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2005年第3期234-237,共4页
Journal of Third Military Medical University
