Zhang Qiang, Lin Lejun, Li Weilong, Lu Guowei, Li Xinna
a Department of Otorhinolaryngology Head and Neck Surgery, The Affiliated Yantai Yuhuangding Hospital of Qingdao University , Yantai , China.
b Department of Nuclear Medicine, The Affiliated Yantai Yuhuangding Hospital of Qingdao University , Yantai , China.
J Recept Signal Transduct Res. 2019 Apr;39(2):146-153. doi: 10.1080/10799893.2019.1638403. Epub 2019 Jul 16.
To investigate the effect of miR-223 on thyroid cancer cells, further to study its potential mechanisms. The difference in miR-223 expression between normal thyroid Nthy-ori3-l cells and thyroid cancer SW579 cells was detected by PCR. The miR-223 overexpression and silencing vector transfection were verified by qRT-PCR. To further investigate the role of miR-223 in AQP-1, the AQP-1 siRNA vector was transfected on the basis of transfection of miR-223 inhibitor vector. The cell proliferation was detected by plate cloning, MTT, and cellular immunofluorescence assays. Cell cycle and apoptosis were detected by flow cytometry. Western blot was used to detect the expression of AQP-1 protein. The expression of miR-223 in SW579 cells was higher than that in normal cells. After transfection with miR-223 mimic, miR-223 expression was increased in SW579 cells. MiR-223 inhibitor transfection can inhibit SW579 cells proliferation, promote apoptosis, and inhibit cell cycle G0/G1 arrest. The SW579 cells proliferation was decreased, and the apoptosis rate was increased after transfection of AQP-1 silencing vector. Compared with the AQP-1 siRNA group, the SW579 cells proliferation rate was further reduced, and the apoptosis rate was significantly increased after co-transfection of miR-223 silencing vector and AQP-1 silencing vector. AQP-1 protein was highly expressed in SW579 cells, and miR-223 inhibitor can down-regulate the expression of APQ-1 protein. The expression AQP-1 protein was significantly reduced after transfected with AQP-1 silencing vector. Inhibition of miR-223 expression could suppress proliferation and promote apoptosis of SW579, and its mechanism is related to down-regulation of APQ-1 protein expression.
为研究miR-223对甲状腺癌细胞的影响，并进一步探讨其潜在机制。采用PCR检测正常甲状腺Nthy-ori3-1细胞与甲状腺癌SW579细胞中miR-223表达的差异。通过qRT-PCR验证miR-223过表达和沉默载体转染。为进一步研究miR-223在水通道蛋白-1（AQP-1）中的作用，在转染miR-223抑制剂载体的基础上转染AQP-1 siRNA载体。采用平板克隆、MTT和细胞免疫荧光检测细胞增殖。通过流式细胞术检测细胞周期和凋亡。采用蛋白质免疫印迹法检测AQP-1蛋白的表达。SW579细胞中miR-223的表达高于正常细胞。转染miR-223模拟物后，SW579细胞中miR-223表达增加。转染miR-223抑制剂可抑制SW579细胞增殖，促进凋亡，并抑制细胞周期G0/G1期阻滞。转染AQP-1沉默载体后，SW579细胞增殖降低，凋亡率增加。与AQP-1 siRNA组相比，共转染miR-223沉默载体和AQP-1沉默载体后，SW579细胞增殖率进一步降低，凋亡率显著增加。AQP-1蛋白在SW579细胞中高表达，miR-223抑制剂可下调APQ-1蛋白的表达。转染AQP-1沉默载体后，AQP-1蛋白表达明显降低。抑制miR-223表达可抑制SW579细胞增殖并促进凋亡，其机制与下调APQ-1蛋白表达有关。
