未折叠蛋白反应参与辣椒素对C6胶质瘤细胞凋亡的机制  

UPR involved in apoptosis induced by capsaicine on glioma U251

作　　者：刘亚萍 董富兴 Yaping Liu;Fuxing Dong(Experimental Teaching Center of Morphology,Xuzhou Medical University,Xuzhou 221004,China;Research Center for Neurobiology,Xuzhou Medical University,Xuzhou 221004,China)

机构地区：[1]徐州医科大学形态学实验教学中心,江苏徐州221004 [2]徐州医科大学神经生物学实验中心,江苏徐州221004

出　　处：《生物技术》2018年第5期492-496,共5页Biotechnology

基　　金：徐州市科技计划项目("自噬参与辣椒素诱导的神经胶质瘤细胞凋亡的机制的研究";No.KC15SH009)

摘　　要：[目的]研究辣椒素对C6胶质瘤细胞的影响及其作用机制。[方法]常规培养C6细胞,MTT法检测细胞存活率。然后将C6细胞分为两组:正常对照组和辣椒素组,采用Western Blot检测Bcl-2、Caspase-3、IRE1、Bip、Chop和pe IF2α的表达;在JNK抑制剂SP600125的作用下,Western Blot法分析P-JNK、IRE1、Chop、Bcl-2和Caspase-3的表达情况。[结果]MTT检测结果显示,辣椒素可以引起C6细胞活力的降低,随着孵育时间的延长,C6细胞的活力逐渐降低,至24h时达到最低,再增加辣椒素的作用时间C6细胞的活力不变。Western Blot结果显示,在辣椒素组中,Bcl-2的表达与对照组(0. 7151±. 0949)相比,表达量降低(0. 4886±0. 1205),而Caspase-3的表达(0. 7555±0. 1155)与对照组(0. 5315±0. 0876)相比升高。并且未折叠蛋白反应(UPR)相关蛋白IRE1、Bip、Chop和p-e IF2α的表达水平与对照组相比均增加,差异有统计学意义。加入JNK抑制剂SP600125后,与辣椒素单独作用相比,IRE1、Chop和Caspase-3蛋白的表达降低,而Bcl-2蛋白的表达水平升高。[结论]辣椒素能够引起C6细胞凋亡和未折叠蛋白反应(UPR)的发生,并且JNK信号通路在其中发挥了重要的作用。[ Objective] To study the effect of capsaicin on C6 glioma cells and its mechanism. [ Methods] C6 ceils were cultured routinely and MTr was used to detect cell viability. Then the C6 cells were divided into 2 groups ： normal control group and capsaicine group,the expressions of Bcl- 2, Caspase- 3, IRE1, Bip, Chop and p- eIF2α were observed by Western Blot. Under the presence of the JNK inhibitor SP600125, the P -JNK, IRE1, Chop, Bcl -2 and Caspase -3 were analyzed by Western Blot. [ Results] MTr assay showed that capsaicin could reduce the activity of C6 cells, and with the extension of incubation time,the activity of C6 cells gradually decreased, but the cells survival remained unchanged after 24h. Western Blot showed that the expression of bcl -2 in capsaicin group was decreased （0. 4886 ＋ 0. 1205 ） compared with the control group （0. 7151 ＋ 0.0949） ,while the expression of caspase -3 was increased （0. 7555 ＋ 0. 1155 ） compared with the control group （0.5315 ＋ 0.0876）. In addition, the expression levels of UPR related proteins IRE1, Bip, Chop and p - elf2 were all increased compared with the control group,and the differences were statistically significant. After adding the JNK inhibitor SP600125, the expressions of IRE1, Chop and Caspase - 3 were reduced compared with the effect of capsaicin alone, while the expression level of bcl -2 was increased. [ Conclusion] Capsaicin could induce apoptosis and UPR of C6 cells, and JNK signaling pathway plays an important role in it.

关 键 词：辣椒素 凋亡 未折叠蛋白反应 JNK 胶质瘤 

分 类 号：R73-3[医药卫生—肿瘤]