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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 study the mutation of mitochondrial DNA (mt DNA) coding region in ovarian cancer cell line SKOV3 / tax. Methods DNA was extracted and sequenced. Mutations of mt DNA gene were analyzed. The number of mt DNA mutations, the type of mutation and the mutation sites were compared between SKOV3 / tax and SKOV3 parental cell lines. Results The mutated hot spots of mt DNA of paclitaxel resistant and sensitive cells were Cytb, ND4 and ND5, respectively. Compared with the sensitive cell lines, mt DNA was found more frequently in SKOV3 / tax resistant cell lines The frequency of multi-gene mutation mainly manifested as A → C, A → G, deletion of A, C, insertion of T, C and so on, but the deletion of A and insertion of T increased obviously, and the mutation position of each gene in drug-resistant and sensitive cell lines Points have the same location. Conclusion The Cytb gene, ND4 gene and ND5 gene in the mt DNA coding region of human ovarian cancer cell line SKOV3 are highly mutated. Mutations in the same site in drug-resistant and susceptible cell lines may be closely related to the occurrence of ovarian cancer. Mutations in drug-resistant cell lines were significantly higher than those in sensitive cell lines, suggesting that increased mutations may be related to drug resistance in ovarian cancer.