小窝蛋白-1对人甲状腺上皮细胞增殖、凋亡及胞内活性氧水平的影响

目的: 探讨小窝蛋白-1（Caveolin-1）对人甲状腺上皮细胞系Nthy-ori 3-1增殖、凋亡和胞内活性氧产生的影响。方法: 将正常人甲状腺上皮细胞（Nthy-ori 3-1）分为实验组和阴性对照组，分别转染 Caveolin-1敲减病毒上清液以及阴性对照病毒上清液，以未转染的Nthy-ori 3-1细胞为空白对照组，应用嘌呤霉素筛选出阳性细胞，qRT-PCR和蛋白质印迹检测3组细胞Caveolin-1的mRNA和蛋白表达；MTT 法检测Caveolin-1敲减后细胞增殖能力变化；流式细胞术检测Caveolin 1敲减后细胞的凋亡率；荧光探针DCFH-DA法检测Caveolin-1敲减后细胞内活性氧水平。结果： Caveolin 1抑制性慢病毒感染后，Nthy-ori 3-1细胞Caveolin 1 mRNA和蛋白水平均显著降低（P均<0.01）。抑制Caveolin-1表达后，Nthy-ori 3-1细胞增殖能力较阴性对照组明显降低，凋亡率明显增加，细胞内活性氧水平明显升高（P 均<0.01）。结论： Caveolin-1表达降低通过增加胞内活性氧水平和细胞凋亡，抑制细胞增殖，介导甲状腺上皮细胞损伤。

Effect of Caveolin 1 on the cell proliferation,apoptosis and ROS level in thyroid follicular cells

Objective: To investigate the effects of knockdown of Caveolin 1 on the cell proliferation, apoptosis and reactive oxygen species (ROS) production in thyroid follicular cells Nthy ori 3 1. Methods: Nthy ori 3 1 cells were divided into the experimental group and negative control group, Caveolin 1 knockdown virus supernatant and negative control virus supernatant were transfected, respectively, and the untransfected Nthy ori 3 1 cells were used as the blank group. After 48 h infection, adding puromycin into the media to select positively infected cells, the expression of Caveolin 1 was determined by qRT PCR and Western blotting. The cell proliferation was detected by MTT, the apoptosis rate was detected by flow cytometry, and the ROS level was detected by fluorescent probe DCFH DA. Results: The qRT PCR and Western blotting results showed that the lentiviral shRNA had a specific knockdown effect on the Cav 1 mRNA and protein levels in Nthy ori 3 1 cells (both P<0.01). Compared with the control group, Caveolin 1 knockdown enhanced apoptosis, ROS production, and inhibited the proliferation of thyroid follicular cells (both P<0.01). Conclusion: Decreased Caveolin 1 expression enhanced apoptosis and ROS production, inhibited the proliferation of thyroid follicular cells, which could play a role in the damage of thyroid follicular cells.