论文部分内容阅读
为了探究葡萄糖调节蛋白94(glucose regulated protein 94,Grp94)干扰与内质网应激(endoplasmic reticulum stress,ERS)介导的凋亡的关系,本研究用衣霉素(tunicamycin,Tm)处理人乳腺癌MCF-7细胞,从而建立ERS体外细胞模型。在应激状态下,设计、合成靶向Grp94的si RNA,通过流式细胞术(flow cytometry,FCM)确定干扰Grp94对细胞凋亡的影响,并由Western blotting检测ERS标志物及未折叠蛋白反应(unfolded protein response,UPR)相关通路分子的表达。结果表明,Grp94被干扰后,细胞应激状态加剧,凋亡率上升;IRE1、p-JNK表达增加,Bcl-2水平降低,说明促凋亡效应可能是通过IRE1-JNK-Bcl-2途径来实现的。本研究结果为Grp94应用于乳腺癌的治疗提供了理论依据与新的思路。
In order to explore the relationship between glucose regulated protein 94 (GRp94) interference and endoplasmic reticulum stress (ERS) -mediated apoptosis, human breast was treated with tunicamycin (Tm) Cancer MCF-7 cells to establish an ERS in vitro cell model. We designed and synthesized si RNA targeting Grp94 under stress and determined the effects of Grp94 on apoptosis by flow cytometry (FCM). The ERS and unfolded proteins were detected by Western blotting (unfolded protein response, UPR) related pathway molecules expression. The results showed that after Grp94 was interfered, the cell stress was aggravated and the apoptosis rate was increased. The expression of IRE1 and p-JNK increased and the level of Bcl-2 decreased, which indicated that the pro-apoptotic effect might be through the IRE1-JNK-Bcl-2 pathway Achieve. The results of this study provide a theoretical basis and new ideas for the treatment of breast cancer Grp94.