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目的:探讨还原型谷胱甘肽在启动蛋白酶体抑制剂诱导甲状腺癌细胞凋亡中的作用。方法:选取4种人甲状腺未分化癌细胞系ARO、FRO、KTC2和8305C,分别设空白对照组、万珂处理组;流式细胞仪(FCM)检测细胞凋亡;测定还原型谷胱甘肽(GSH)含量、谷氨酸半胱氨酸合成酶(GCL)和谷胱甘肽合成酶(GS)的活性;实时定量PCR法检测GCL和GS mRNA的水平。结果:与对照组相比,蛋白酶体抑制剂万珂对人甲状腺癌细胞系FRO和KTC2具有很强的细胞凋亡诱导作用;万珂处理24h时,FRO、KTC2细胞中GSH水平下降,差异有统计学意义,P<0.01;ARO和8305C细胞中GSH水平显著增加,差异有统计学意义,P<0.01。万珂处理组FRO和KTC2甲状腺癌细胞中,GCL的活性没有变化而GCL mRNA也仅有轻度增加,P>0.05;但在ARO和8305C细胞中,万珂作用8h后,GCL的活性及mRNA的表达显著增加,差异有统计学意义,P<0.01;而GS活性及mRNA水平在各组细胞中的变化差异均无统计学意义,P>0.05。结论:蛋白酶体抑制剂可诱导耐药的甲状腺癌细胞中GCL转录和活性增高,提高GSH在细胞中的表达,导致蛋白酶体抑制剂诱导的肿瘤细胞凋亡受抑。
Objective: To investigate the role of reduced glutathione in the activation of proteasome inhibitor-induced apoptosis of thyroid cancer cells. Methods: Four kinds of human thyroid undifferentiated carcinoma cell lines ARO, FRO, KTC2 and 8305C were selected and divided into blank control group and Velcade group. Flow cytometry (FCM) (GSH), glutamate cysteine synthase (GCL) and glutathione synthetase (GS) activity, and real-time quantitative PCR method was used to detect the levels of GCL and GS mRNA. Results: Compared with the control group, the proteasome inhibitor Velcade had a strong apoptosis-inducing effect on human thyroid cancer cell lines FRO and KTC2; and when treated by Velcro for 24 hours, the levels of GSH in FRO and KTC2 cells decreased Statistical significance, P <0.01; ARO and 8305C cells GSH levels increased significantly, the difference was statistically significant, P <0.01. There was no change in the activity of GCL and the slight increase of GCL mRNA in FRO and KTC2 thyroid cancer cells treated with Velcade, P> 0.05; however, in ARO and 8305C cells, the activity of GCL and mRNA The difference was statistically significant (P <0.01). However, there was no significant difference in the changes of GS activity and mRNA between the two groups (P> 0.05). CONCLUSION: Proteasome inhibitor can induce the increase of GCL transcription and activity in drug-resistant thyroid carcinoma cells and increase the expression of GSH in the cells, leading to inhibition of tumor cell apoptosis induced by proteasome inhibitor.