PKM2影响鼻咽癌细胞增殖凋亡及机制研究  

Effect and mechanism of PKM2 on proliferation and apoptosis of nasophar-yngeal carcinoma cells

作　　者：丁元平 孙丽 郝妮妮 远洋[1] 赵立民[1] DING Yuan-Ping;SUN Li;HAO Ni-Ni;YUAN Yang;ZHAO Li-Min(Department of Otorhinolaryngology, Affiliated Hospital of Weifang Medical University, Weifang 261031, China)

机构地区：[1]潍坊医学院附属医院耳鼻咽喉科,潍坊261031

出　　处：《中国免疫学杂志》2018年第5期675-680,共6页Chinese Journal of Immunology

摘　　要：目的:探讨PKM2对鼻咽癌细胞增殖凋亡的影响。方法:鼻咽癌细胞CNE-1转染PKM2小干扰RNA(PKM2siRNA1和PKM2 siRNA2)和阴性对照(siRNA control),荧光定量PCR和Western blot检测细胞中PKM2水平,筛选干扰效果好的PKM2 siRNA2继续研究。噻唑蓝(MTT)检测细胞增殖,细胞克隆试验检测细胞克隆形成能力,流式细胞术检测细胞凋亡,二氯二氢荧光素-乙酰乙酸酯(DCFH-DA)法检测活性氧(ROS)水平,Western blot检测p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化的p38MAPK(p-p38MAPK)、C-myc、β-连环蛋白(β-catenin)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)蛋白水平。结果:细胞转染PKM2 siRNA1和PKM2 siRNA2后PKM2 mRNA和蛋白水平与没有转染的细胞相比均明显下降,并且转染PKM2 siRNA2后细胞中PKM2水平下降更多,而转染siRNA control的细胞中PKM2水平与没有转染的细胞相比没有明显变化。下调PKM2表达后的细胞凋亡率由(9.36±1.04)%升高至(48.42±5.28)%,细胞克隆形成率从(75.48±8.25)%降低至(46.15±3.47)%,细胞OD值从(0.86±0.11)下降至(0.52±0.04),细胞中ROS水平升高,细胞中p-p38MAPK、Cleaved Caspase-3蛋白水平也明显升高,细胞中C-myc、β-catenin水平明显下降。结论:PKM2表达下调抑制鼻咽癌细胞生长,促进鼻咽癌细胞凋亡,作用机制可能与p38MAPK和Wnt/β-catenin信号通路有关。Objective： To investigate the effect of PKM2 on proliferation and apoptosis of nasopharyngeal carcinoma cells. Methods： Nasopharyngeal carcinoma cell CNE-1 was transfected with PKM2 small interfering RNA（ PKM2 siRNA1 and PKM2 siRNA2） and negative controls（ siRNA control）,the levels of PKM2 in the cells were detected by fluorescent quantitative PCR and Western blot,screening interference PKM2 siRNA2 continued to study. Cell proliferation was detected by MTT,cell cloning test showed the ability of cell cloning,apoptosis was detected by flow cytometry,ROS level was detected by DCFH-DA,the levels of p38 MAPK,pp38 MAPK,C-myc,β-catenin,Cleaved Caspase-3 protein were detected by Western blot. Results： After transfection of PKM2 siRNA1 and PKM2 siRNA2,the levels of PKM2 mRNA and protein were significantly decreased compared with those without transfection,and after transfection of PKM2 siRNA2,the level of PKM2 in cells decreased more,the levels of PKM2 in transfected siRNA control cells were not significantly different from those without transfection. The rate of apoptosis after down-regulation of PKM2 expression increased from（ 9. 36 ± 1. 04） % to（ 48. 42 ± 5. 28） %,and the rate of cell clone formation decreased from（ 75. 48 ± 8. 25） % to（ 46. 15 ±3. 47） %,OD values from（ 0. 86 ±0. 11） down to（ 0. 52 ±0. 04）,elevated levels of ROS in cells,the levels of p-p38 MAPK,Cleaved Caspase-3 proteins in cells were also significantly increased,the levels of C-myc and β-catenin in cells were obviously decreased. Conclusion： Downregulation of PKM2 expression inhibits nasopharyngeal carcinoma cell growth,promoting apoptosis of nasopharyngeal carcinoma cells,the mechanism of action may be related to p38 MAPK and Wnt/β-catenin signaling pathway.

关 键 词：鼻咽癌 PKM2 增殖 凋亡 

分 类 号：R739.63[医药卫生—肿瘤]