摘要
目的探讨碱性螺旋-环-螺旋转录因子1(Twist basic helix-loop-helix transcription factor 1,Twist1)与Notch信号通路配体Jagged1(Notch ligand Jagged-1,Jagged1)在人卵巢癌A2780细胞顺铂(cisplatin,DDP)耐药中的作用。方法实验分为敏感组(人卵巢癌DDP敏感A2780细胞),耐药组(DDP耐药A2780/DDP细胞),转染组[应用小分子干扰RNA(small interfering RNA,siRNA)技术转染A2780/DDP细胞,干扰Twist1基因表达为A2780/DDP/si-Twist1、干扰Jagged1基因表达为A2780/DDP/si-Jagged1、空转不干扰Twist1及Jagged1基因表达为A2780/DDP/si-NC]。采用Western blot法检测3组Twist1、Jagged1蛋白表达水平;采用CCK-8比色法检测耐药组和转染组细胞DDP敏感性,生长曲线分析耐药组和转染组细胞增殖状况,流式细胞仪检测耐药组和转染组细胞凋亡情况。结果耐药组A2780/DDP细胞Twist1和Jagged1蛋白表达水平均高于敏感组A2780细胞(P<0.05);转染组A2780/DDP/si-Twist1细胞Twist1和Jagged1蛋白表达水平均低于耐药组A2780/DDP细胞和转染组A2780/DDP/si-NC细胞(P<0.05);转染组A2789/DDP/si-Jagged1细胞Jagged1蛋白表达水平低于耐药组A2780/DDP细胞和转染组A2780/DDP/si-NC细胞(P<0.05),Twist1蛋白表达水平与耐药组A2780/DDP细胞和转染组A2780/DDP/si-NC细胞比较差异无统计学意义(P>0.05);转染组A2780/DDP/si-Twist1细胞对DDP的半数抑制浓度(half maximal inhibitory concentration,IC50)[(19.53±0.32)mg/L]低于耐药组A2780/DDP细胞[(30.57±0.27)mg/L]和转染组A2780/DDP/si-NC细胞[(29.61±0.35)mg/L],细胞凋亡率[(34.74±3.15)%]高于耐药组A2780/DDP细胞[(21.56±1.99)%]和转染组A2780/DDP/si-NC细胞[(23.49±2.27)%](P<0.05)。结论 Twist1可通过调控Jagged1基因表达,介导人卵巢癌A2780细胞的增殖和凋亡,参与对DDP耐药的调节。
Objective To explore the effects of Twist basic helix-loop-helix transcription factor 1(Twist1)and Notch ligand Jagged-1(Jagged1)genes on the resistance of human ovarian cancer A2780 cells to cisplatin(DDP).Methods The experiment included DDP-sensitive human ovarian cancer A2780 cells(DDP-sensitive group),DDP-resistant A2780/DDP cells(DDP-resistant group),and A2780/DDP cells transfected by small interfering RNA(siRNA)(transfection group).In transfection group,A2780/DDP/si-Twist1 cells and A2780/DDP/si-Jagged1 cells were obtained by interfering the expressions of Twist1 and Jagged1 genes in A2780/DDP cells respectively,and A2780/DDP/si-NC cells were not interfered.The expression levels of Twist1 and Jagged1 proteins were detected by Western blot method in three groups.The sensitivity of DDP was detected by CCK-8 method in DDP-resistant group and transfection group.The cell proliferation was analyzed with growth curve and the apoptosis was detected by flow cytometry in DDP-resistant group and transfection group.Results The expression levels of Twist1 and Jagged1 proteins were significantly higher in A2780/DDP cells in DDP-resistant group than those in A2780 cells in DDP-sensitive group(P<0.05).The expression levels of Twist1 and Jagged1 proteins were significantly lower in A2780/DDP/si-Twist1 cells in transfection group than those in A2780/DDP cells in DDP-resistant group and in A2780/DDP/si-NC cells in transfection group(P<0.05).The expression of Jagged1 protein was significantly lower in A2780/DDP/si-Jagged1 cells in transfection group than those in A2780/DDP cells in DDP-resistant group and in A2780/DDP/si-NC cells in transfection group(P<0.05),and the expression of Twist1 protein in 2780/DDP/si-Jagged1 cells in transfection group showed no significant differences in comparison with that in A2780/DDP cells in DDP-resistant group and A2780/DDP/si-NC cells in transfection group(P>0.05).The half maximal inhibitory concentration of A2780/DDP/si-Twist1 cells to DDP was significantly lower in transfection group((19.53±0.32)mg/L)than that of A2780/DDP cells in DDP-resistant group((30.57±0.27)mg/L)and of A2780/DDP/si-NC cells in transfection group((29.61±0.35)mg/L),while DDP-induced apoptosis rate of A2780/DDP/si-Twist1 cells in transfection group((34.74±3.15)%)was significantly higher than that of A2780/DDP cells in DDP-resistant group((21.56±1.99)%)and of A2780/DDP/si-NC cells in transfection group((23.49±2.27)%)(P<0.05).Conclusion Twist1 protein could contribute to the resistance of A2780 cells to DDP in ovarian cancer by regulating the expression of Jagged1 and mediating the proliferation and apoptosis of A2780 cells.
出处
《中华实用诊断与治疗杂志》
2017年第11期1067-1071,共5页
Journal of Chinese Practical Diagnosis and Therapy
基金
湖北省医学领军人才培养工程专项经费资助
湖北省卫生和计划生育委员会重点项目(WJ2015MA024)
