腺相关病毒介导的CDK2-shRNA促进黑色素瘤细胞A375凋亡

发布时间：2019-06-13 22:54

【摘要】：目的探讨腺相关病毒介导的CDK2-shRNA对人黑色素瘤细胞A375凋亡,为黑色素瘤的防治提供技术支持。方法体外合成CDK2及转录终止序列,构建CDK2-shRNA的腺相关病毒载体和包装相应的重组腺相关病毒,荧光显微镜检测转染和感染条件下CDK2-shRNA表达效果,MTT法检测细胞活力,Annexin V-FITC/PI细胞凋亡试剂盒检测细胞凋亡,免疫印迹检测cyclin E和E2F1表达。结果转染和感染的腺相关病毒介导的CDK2-shRNA在A375细胞均有表达且具有较低的细胞毒性;转染和感染的CDK2-shRNA均促进A375细胞发生凋亡;干扰CDK2可降低cyclin E和E2F1表达。结论腺相关病毒介导的CDK2-shRNA促进黑色素瘤细胞A375凋亡。
[Abstract]:Objective to investigate the apoptosis of human melanoma cell line A375 mediated by adeno-associated virus (CDK2-shRNA), and to provide technical support for the prevention and treatment of melanoma. Methods CDK2 and transcriptional termination sequence were synthesized in vitro. The adeno-associated virus vector of CDK2-shRNA and the corresponding recombinant adeno-associated virus were constructed. The expression of CDK2-shRNA was detected by fluorescence microscope, the cell vitality was detected by MTT assay, the apoptosis was detected by Annexin V-FITC/PI apoptosis kit, and the expression of cyclin E and E2F1 was detected by immunoblotting. Results both transfected and infected adeno-associated virus-mediated CDK2-shRNA were expressed in A375 cells and had low cytotoxicity. Both transfected and infected CDK2-shRNA promoted apoptosis of A375 cells. Interference with CDK2 could decrease the expression of cyclin E and E2F1. Conclusion adeno-associated virus mediated CDK2-shRNA promotes apoptosis of melanoma cell line A375.
【作者单位】：黑龙江中医药大学;福建中医药大学附属厦门第三医院皮肤科;吉林大学中日联谊医院普通外科;
【基金】：厦门市卫生局资助项目(3502Z20130190) 厦门市同安区科技局资助项目
【分类号】：R739.5

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