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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 investigate the effect of Jagged1 (Notch ligand Jagged-1, Twist basic helix-loop-helix transcription factor 1) and cisplatin on human ovarian cancer cell line A2780 (cisplatin, DDP) resistance role. Methods The experiment was divided into two groups: sensitive group (DDP sensitive A2780 cell line), drug resistant group (DDP resistant A2780 / DDP cell line), transfected group [transfected by A2780 / DDP cells, which interfere with the expression of Twist1 gene in A2780 / DDP / si-Twist1 and interfere with the expression of Jagged1 gene in A2780 / DDP / si-Jagged1, without interfering with Twist1 and Jagged1 gene expression in A2780 / DDP / si-NC. Western blot was used to detect the expression of Twist1 and Jagged1 in each group. CCK-8 colorimetric assay was used to detect the DDP sensitivity of the drug-resistant group and the transfection group. The growth curve was used to analyze the cell proliferation status in the drug-resistant group and the transfected group. Cytometry was used to detect cell apoptosis in drug-resistant and transfection groups. Results The expression of Twist1 and Jagged1 in A2780 / DDP cells was higher than that in A2780 cells (P <0.05). The expression of Twist1 and Jagged1 in A2780 / DDP / si-Twist1 cells were lower than that in A2780 cells A2780 / DDP cells and A2780 / DDP / si-NC cells (P <0.05). The expression of Jagged1 protein in A2789 / DDP / si-Jagged1 cells was lower than that in A2780 / DDP cells and transfection group The expression of Twist1 protein in A2780 / DDP / si-NC cells was not significantly different from that in A2780 / DDP / si-NC cells (P> 0.05). The half maximal inhibitory concentration (IC50) [(19.53 ± 0.32) mg / L] in transfected A2780 / DDP / si-Twist1 cells was lower than that in A2780 / DDP cells [(30.57 ± 0.27) (29.61 ± 0.35) mg / L], and the apoptosis rate [(34.74 ± 3.15)%] was higher in A2780 / DDP cells transfected with A2780 / DDP / 21.56 ± 1.99)%] and the transfected group A2780 / DDP / si-NC cells [(23.49 ± 2.27)%] (P <0.05). Conclusion Twist1 can regulate the expression of Jagged1 gene and mediate the proliferation and apoptosis of human ovarian cancer cell line A2780, which is involved in the regulation of DDP resistance.