摘要
[目的]探讨β-榄香烯(β-elemene,β-ELE)对人脑胶质瘤细胞株U251凋亡的诱导作用及对c-myc、hTERT、microRNA分子表达的影响。[方法]用β-榄香烯处理U251细胞株24小时,流式细胞术检测细胞凋亡;RT-PCR法检测c-myc、hTERT基因的表达;Western-blot检测cmyc、hTERT蛋白的表达;基因芯片检测β-榄香烯处理对细胞表达microRNA的影响,并以Real time-PCR法验证其中miR-654-3p、miR-431、miR-150和miR-1976的表达水平。[结果]β-榄香烯明显诱导U251细胞的凋亡,与对照组(0μg·ml-1β-ELE)相比,实验组(100,200,300μg·ml-1β-ELE)的细胞凋亡有所增加,凋亡程度随着β-ELE的浓度增大而增加,具有统计学差异;β-榄香烯抑制了c-myc、hTERT基因mRNA和蛋白的表达,与对照组(0μg·ml-1β-ELE)相比,实验组(50,100,150μg·ml-1β-ELE)的c-myc、hTERT基因mRNA和蛋白的表达水平都有所下降,下降程度随着β-ELE的浓度增大而增加,具有统计学差异;基因芯片和Real time-PCR结果显示,与对照组(0μg·ml-1β-ELE)相比,实验组(150μg·ml-1β-ELE)miR-654-3p、miR-431、miR-150和miR-1976的表达有所上调,具有统计学差异。[结论]β-榄香烯诱导了U251细胞凋亡抑制了c-myc、hTERT基因mRNA与蛋白的表达,显著上调了miR-654-3p、miR-431、miR-150和miR-1976的表达水平。因此,β-榄香烯诱导U251细胞凋亡的作用可能是通过其抑制c-myc、hTERT基因mRNA和蛋白的表达及上调microRNA来实现的。此机制为抗肿瘤药物的发展提供了一定的理论基础,值得进一步深入探讨。
[Objective]To investigate the effects of β-elemene on apoptosis of human glloma U251 cells and on the expression of c-myc,hTERT and microRNA. [Methods] U251 cells were treated with β-elemene or the vehicle for 24 hours, followed by the apoptotic analysis with flow cytometry, and qualitative test for the c-myc, hTERT levels with RT-PCR and Western-blot. Furthermore, expression of microRNA was analyzed by gene microarray, and the selective microRNAs including miR-654-3p,miR-431,miR-150 and miR-1976 were examined by Real time-PCR. [Results] β-elemene obviously induced U251 cell apoptosis, which was associated with dose-dependent reduction in expression of c--myc, hTERT, at mRNA and protein levels. Microarray and Real time-PCR analysis showed that ]3-elemene altered the expression of miR-654-3p,miR-431,miR-150 and miR-1976 in the glioma. [Conclusions] β-elemene induces U251 apoptosis, inhibits the expressions of c-myc, hTERT, gene mRNA and protein, and markedly up- regulates the expression of miR-654-3p,miR-431,miR-150 and miR-1976, therefore, the function of β-elernene inducing U251 apoptosis may have the mechanism of inhibiting the expression of c-myc, hTERT, gene mRNA and protein and up-regulating microRNA, which offers definite theoretic foundation for developing anti-tumor drugs, worth further discussion.
出处
《浙江中医药大学学报》
CAS
2014年第4期451-455,460,共6页
Journal of Zhejiang Chinese Medical University
基金
国家自然科学基金项目(81270066)
教育部优博作者专项基金项目(201070)
浙江省自然科学基金项目(LY12H10007)
杭州市医疗科研及重点专科专病项目(20100633B11)~~
