龙葵碱联合KLF16基因对胶质瘤细胞增殖、凋亡的影响及其机制研究  

Solanum nigrum combined with KLF16 gene synergistically inhibit glioma cell proliferation and induce apoptosis

作　　者：赵舒杨 雷艳杰 马世杰[1] 高明[1] ZHAO Shuyang;LEI Yanjie;MA Shijie;GAO Ming(Department of Neurosurgery,Zhoukou Central Hospital,Zhoukou,Henan,China,466000;Department of Pathology,Zhoukou Central Hospital,Zhoukou,Henan,China,466000)

机构地区：[1]周口市中心医院神经外科,河南周口466000 [2]周口市中心医院病理科,河南周口466000

出　　处：《分子诊断与治疗杂志》2019年第4期303-309,共7页Journal of Molecular Diagnostics and Therapy

摘　　要：目的探讨龙葵碱联合Krüppel样转录因子16(KLF16)基因对人胶质瘤U87细胞增殖和凋亡的影响及作用机制。方法体外培养人胶质瘤U87细胞,将特异性KLF16 siRNA和阴性对照siRNA Control转染U87细胞,分别命名为si-KLF16组和si-NC组,应用实时荧光定量PCR(qRT-PCR)和蛋白免疫印迹(Western blot)检测转染后细胞中KLF16的表达情况。使用不同浓度(2.5、5、10、20、30μg/μL)的龙葵碱分别作用si-NC组和si-KLF16组细胞48 h,噻唑蓝(MTT)法测定细胞增殖抑制率并筛选半数抑制浓度(IC50),以IC50为龙葵碱后续实验浓度,根据是否添加龙葵碱处理实验分为4组,si-NC组、si-KLF16组、si-NC+Sol组、si-KLF16+Sol组,分别使用MTT和流式细胞仪测定各组细胞增殖和凋亡能力,Western blot分析各组细胞中Ki-67、PCNA、Bax和Bcl-2蛋白表达情况。结果胶质瘤U87细胞转染后,si-KLF16组细胞中KLF16 mRNA和蛋白表达显著低于si-NC组(P<0.05)。筛选出浓度为20μg/μL的龙葵碱作为后续实验浓度。与si-NC组比,si-KLF16组、si-NC+Sol组和si-KLF16+Sol组细胞增殖能力均受到明显抑制(P<0.05),凋亡率升高(P<0.05),Ki-67、PCNA和Bcl-2蛋白表达下调(P<0.05),Bax蛋白表达上调(P<0.05)。与si-KLF16组和si-NC+Sol组比,si-KLF16+Sol组细胞增殖抑制作用和凋亡促进作用更显著(P<0.05)。结论龙葵碱联合KLF16基因能够协同抑制胶质瘤细胞增殖,诱导细胞凋亡,其作用机制可能与调控Ki-67、PCNA、Bax和Bcl-2蛋白表达有关。Objective To investigate the effects of solanine combined with Krüppel-like transcription factor 16(KLF16)gene on proliferation and apoptosis of human glioma U87 cells. Methods Human glioma U87 cells were cultured in vitro,and specific KLF16 siRNA and negative control siRNA Control were transfected into U87 cells,named si-KLF16 group and si-NC group,respectively. The expression of KLF16 in transfected cells was detected by real-time quantitative PCR(qRT-PCR)and Western blot. The cells of si-NC group and si-KLF16 group were treated with different concentrations(2.5,5,10,20,30 μg/μL)of solanine for 48 h. The inhibition rate of cell proliferation was determined by MTT assay and half of the cells were screened. Inhibitory concentration(IC50),IC50 is the subsequent experimental concentration of Solanum. Divided into four groups according to whether or not to add the solanine treatment experiment,siNC group,si-KLF16 group,si-NC+Sol group,si-KLF16+Sol group. The proliferation and apoptosis of each group were determined by MTT and flow cytometry. The expressions of Ki-67,PCNA,Bax and Bcl-2 in each group were analyzed by Western blot. Results After transfection of glioma U87 cells,the expression of KLF16 mRNA and protein in si-KLF16 group was significantly lower than that in si-NC group(P<0.05).Solanine at a concentration of 20 μg/μL was selected as the subsequent experimental concentration. Compared with the si-NC group,the cell proliferation ability of the si-KLF16 group,the si-NC+Sol group and the siKLF16+Sol group were significantly inhibited(P<0.05),and the apoptotic rate was increased(P<0.05). Ki-67,PCNA and Bcl-2 protein expression was down-regulated(P<0.05),Bax protein expression was up-regulated(P<0.05). Compared with the si-KLF16 group and the si-NC + Sol group,the cell proliferation inhibition and apoptosis promotion effects of the si-KLF16+Sol group were more significant(P<0.05). Conclusion Solanum nigrum combined with KLF16 gene can synergistically inhibit glioma cell proliferation and induce apoptosis,w

关 键 词：龙葵碱 KLF16 基因 胶质瘤U87 细胞 

分 类 号：R285.5[医药卫生—中药学]