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目的:明确Nrf2对蛋白酶体抑制剂万珂诱导甲状腺癌细胞凋亡的影响及其作用途径。方法:选取Nrf2高表达的8305C细胞,用微小RNA干扰技术转染细胞,采用实时定量PCR法和蛋白质印迹法分析封闭效果,而细胞内GCLCmRNA的表达、还原型谷胱甘肽(GSH)的含量和细胞凋亡率,分别由实时定量PCR法、比色法和流式细胞仪法测得。并测定siNrf2对万珂诱导其他甲状腺癌细胞凋亡的影响。结果:与对照组相比,siNrf2组在培养液和万珂处理中Nrf2mRNA和蛋白都显著减少,P<0.01;随机序列核酸siRNA和错位型siNrf2差异无统计学意义,P>0.05。与对照组相比,siNrf2能够增加万珂诱导8305C、8505C、KTC1和KTC3的细胞凋亡率,差异均有统计学意义,P<0.01;FRO和KTC2的细胞凋亡率差异无统计学意义,P>0.05。与随机序列核酸siRNA组相比,空白对照组中siNrf2转染组GCLCmRNA及GSH减少,P<0.05;万珂处理组中siNrf2转染组两者减少更显著,P<0.01。GSH和NAC(GSH的前体并能提高GSH的含量)抑制了siNrf2促万珂诱导甲状腺癌细胞凋亡作用。结论:Nrf2抵消万珂诱导甲状腺癌细胞凋亡作用,至少部分是通过GCLC转录激活和随后GSH的生成实现的。
Objective: To investigate the effect of Nrf2 on the apoptosis of thyroid carcinoma cells induced by the proteasome inhibitor Viaduct and its pathway. Methods: The Nrf2-overexpressing 8305C cells were selected and transfected into cells using microRNA interference technique. The real-time quantitative PCR and Western blotting were used to analyze the effect of encapsulation. However, the expression of GCLCmRNA, the content of reduced glutathione (GSH) And apoptosis rate were measured by real-time quantitative PCR, colorimetric and flow cytometry. And determine the effect of siNrf2 on the apoptosis of other thyroid cancer cells induced by Velcro. Results: Compared with the control group, the Nrf2 mRNA and protein in the siNrf2 group were significantly decreased (P <0.01). The difference between the siRNA sequence of random sequence and the siNrf2 dislocation was not significant (P> 0.05). Compared with the control group, siNrf2 increased the apoptosis rate of 8305C, 8505C, KTC1 and KTC3 induced by Velocity, the difference was statistically significant, P <0.01; There was no significant difference in apoptosis rate between FRO and KTC2, P> 0.05. Compared with the siRNA group with random sequence, GCLC mRNA and GSH decreased in the siNrf2 transfected group (P <0.05) in the blank control group; siNrf2 transfected group decreased more significantly (P <0.01). GSH and NAC (precursors to GSH and increase GSH content) inhibit the apoptosis of thyroid cancer cells induced by siNrf2-Enke. CONCLUSION: Nrf2 counteracts the atorvastatin-induced apoptosis of thyroid cancer cells, at least in part by GCLC transcriptional activation and consequent GSH production.