死亡结构域相关蛋白对卵巢癌细胞周期及化疗影响的研究

Study on influence of Daxx on cell cycle and chemotherapy in human ovarian cancer cells

目的探索在卵巢癌细胞中下调死亡结构域相关蛋白(Daxx)后对其细胞周期及化疗耐药的影响。方法转染卵巢癌耐药细胞株C13k后分为阴性对照组(NC组)和下调Daxx组(siDaxx组)。设立多柔比星药物浓度梯度0、0.15、0.30、0.60μmol/L,流式细胞仪检测上述两组细胞周期变化。Western blot从蛋白水平检测细胞周期及凋亡相关蛋白cyclinB1、cleaved-parp的表达变化。流式细胞仪检测上述两组细胞凋亡的变化。结果在多柔比星0.30μmol/L时,下调Daxx蛋白使细胞发生明显的G2/M期阻滞,与NC组比较,siDaxx组G2/M期细胞百分比明显增高(P<0.05)。下调Daxx蛋白使细胞对多柔比星化疗耐药。结论 Daxx对卵巢癌化疗的影响可能与Daxx对细胞周期的调控作用有关。

Objective To explore the influence of down-regulating l）axx on cell cycle and chemotherapeutic drug resistance in human ovarian cancer cells. Methods SiRNA and NC negative eontrol（NC） RNA of Daxx were constructed and divided into the NC group and silencing Daxx（siDaxx） group after transfecting to ovarian cancer cell line C13k. The doxorubicin concentration gra- dient of 0,0.15,0.30,0.60 μmol/L was set. The cellular cycle changes and apoptosis changes of these two groups were detected by using the flow cytometry. Western blot was used to detect the expression changes of apoptosis related protein cyclinB1 and cleaved- parp. Results 0. 30 μmol/L doxorubicin down-regulated Daxx to result in significant G2/M arrest （P〈0. 05）. Down-regulating Daxx resulted in doxorubicin resistance in C13k cells. Conclusion The effect of Daxx on ovarian cancer chemotherapy might be related to its regulation on cell cycle.