卵巢癌紫杉醇耐药细胞株SKOV3/tax中mtDNA基因突变的检测

The detection of mitochondrial DNA mutation in taxol-resistant ovary cancer cell line SKOV3/tax

目的研究卵巢癌紫杉醇耐药细胞株SKOV3/tax中线粒体DNA(mt DNA)编码区部分基因的突变。方法提取细胞DNA并测序,进行mt DNA基因突变的分析,比较SKOV3/tax与亲本敏感细胞SKOV3之间mt DNA基因突变数量、突变类型、突变位点的差异。结果在检测的基因中,紫杉醇耐药及敏感细胞mt DNA的突变热点依次是Cytb基因、ND4基因和ND5基因,与敏感细胞株相比,耐药细胞株SKOV3/tax中mt DNA测序中发现更多基因突变频率,主要表现为A→C,A→G,缺失A、C,插入T、C等,而缺失A,插入T等明显增加,耐药及敏感细胞株中每个基因的突变位点均有相同位置。结论人卵巢癌细胞株SKOV3中mt DNA编码区的Cytb基因、ND4基因和ND5基因是具有高度突变性的区域,耐药及敏感细胞株中相同位点突变可能与卵巢癌的发生有重要关系。耐药细胞株中的突变明显高于敏感细胞株,推测突变的增加可能与卵巢癌耐药有关。

Objective： To explore the mutation of mtDNA coding region in Taxol-resistant ovary cancer cell line SKOV3/tax. Methods： mtDNA were extracted and sequencing .The mutation was analyzed in the Taxol-resistant ovary cancer cell line SKOV3/ tax and their parent cell lines SKOV3. The difference of mtDNA mutation rate and type were compared between them. Results： In the detected gene, the hot point mutation were Cytb gene, ND4 gene and ND5 gene in subsequently. Compare with the sensitive cell line, there were more changes in genetic mutations in drug-resistent one. The main mutation types were A→C, A→G, lost A/ C, insert T/C. And lost A, insert T is the remarkable characteristics. Conclusions： The mtDNA gene of Cytb, ND4, NDS, are the highly mutation region in ovary cancer cell line, which may be have some relations with ovary carcinogenesis and development. MtDNA mutation in drug-resistent cell line have more marked difference than in the sensitive one, indicating that may be related with the mechanism of Taxol-resistance in ovarian cancer.