摘要
目的探讨微小RNA-378a-3p(miR-378a-3p)与PKM2基因的关系,以及miR-378a-3p对食管鳞癌细胞Eca-109凋亡及能量代谢的影响.方法采用Eca-109细胞系,后续实验分为空白对照组、阴性对照组(转染negative control)和实验组(miR-378a-3p基因转染24 h与48 h).利用流式细胞技术检测miR-378a-3p转染后24 h和48 h对Eca-109细胞凋亡的影响;用紫外分光光度法检测miR-378a-3p转染后24 h和48 h对Eca-109细胞能量代谢的影响;用蛋白免疫印迹法(Western blot-ting)检测miR-378a-3p干预后PKM2蛋白水平的变化.结果与空白对照组和阴性对照组相比,实验组miR-378a-3p转染24 h和48 h时PKM2蛋白的表达水平降低(P<0.05),且48 h较24 h时PKM2蛋白的表达水平降低更明显.与空白对照组和阴性对照组相比,实验组转染miR-378a-3p后24 h和48 h,紫外分光光度法检测ATP含量明显降低(P<0.01),代谢减少,且48 h较24 h的ATP产量降低更明显.与空白对照组和阴性对照组相比,实验组miR-378a-3p转染后24 h和48 h Eca-109细胞的凋亡率显著增高(P<0.001),48 h较24 h时细胞凋亡率增高更明显.结论miR-378a-3p通过抑制有氧酵解关键酶PKM2来干预食管癌细胞的能量代谢,进而增加细胞的凋亡.
Objective To investigate the relationship between microRNA-378a-3p(miR-378a-3p)and PKM 2 gene,and the effect of miR-378a-3p on the apoptosis of esophageal squamous cell carcinoma Eca-109 and the energy metabolism.Methods The Eca-109 cell line was used and divided into blank control group,negative control group(transfected with negative control)and experimental group(transfected with miR-378a-3p gene for 24 h and 48 h).Effect of miR-378a-3p on the apoptosis of Eca-109 cells at 24 h and 48 h was detected by flow cytometry.Effect of miR-378a-3p on the energy metabolism of Eca-109 cells at 24 h and 48 h was detected by UV spectrophotometry.Changes of PKM 2 protein levels after miR-378a-3p intervention was detected by Western blotting.Results Compared with blank control group and negative control group,the expression level of PKM 2 protein was significantly reduced at 24 h and 48 h after miR-378a-3p transfection in experimental group(P<0.05),and the expression level of PKM 2 protein was significantly lower at 48 h than at 24 h.Compared with blank control group and negative control group,the content of ATP was significantly reduced and the metabolism was decreased by ultraviolet spectrophotometry at 24 h and 48 h after transfection with miR-378a-3p in experimental group(P<0.01),and the ATP production was lower at 48 h than at 24 h.Compared with blank control group and negative control group,the apoptosis rate of Eca-109 cells was significantly increased at 24 h and 48 h after miR-378a-3p transfection in experimental group(P<0.001),and the apoptosis rate was higher at 48 h than at 24 h.Conclusion The miR-378a-3p blocks the energy metabolism of esophageal cancer cells by inhibiting the aerobic key enzyme PKM 2 thereby increasing cell apoptosis.
作者
阿依提拉·热合麦提江
屈园
帕力旦·艾孜提
张法煌
朱世茂
胡慧玲
李卉
REHEMAITIJIANG Ayitila;QU Yuan;AIZITI Palidan;ZHANG Fahuang;ZHU Shimao;HU Huiling;LI Hui(Department of Biochemistry and Molecular Biology,Basic Medical College,Xinjiang Medical University,Urumqi 830011,China;College of Public Health,Xinjiang Medical University;Central Laboratory,Xinjiang Medical University)
出处
《山西医科大学学报》
CAS
2020年第4期283-287,共5页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81660459)。
