新疆维吾尔自治区人民医院 乳腺甲状腺科，新疆 乌鲁木齐，830001
Xinjiang Uygur Autonomous Region People's Hospital, Urumqi, 830001, Xinjiang, China
目的 探讨环状RNA 0000064（circ_0000064）靶向微小RNA 503（miR-503）对甲状腺癌细胞生物学行为的影响。方法 实时定量PCR（RT-qPCR）分析21例甲状腺癌组织和对应癌旁组织中circ_0000064和miR-503的表达水平。将circ_0000064小干扰RNA（si-circ_0000064）、miR-503模拟物、si-circ_0000064+miR-503抑制物（anti-miR-503）分别转染甲状腺癌SW579细胞，采用平板克隆形成实验、细胞计数试剂盒（CCK-8）法检测SW579细胞增殖，Transwell实验检测SW579细胞迁移和侵袭，流式细胞术检测SW579细胞凋亡，蛋白质印迹法检测上皮型钙黏蛋白（E-cadherin）和神经型钙黏蛋白（N-cadherin）表达量。双荧光素酶报告实验和RT-qPCR分析circ_0000064和miR-503的靶向调控关系。结果 甲状腺癌组织中circ_0000064表达量显著高于癌旁组织（P<0.05），miR-503表达量显著低于癌旁组织（P<0.05）。沉默circ_0000064后SW579细胞活力、克隆形成数、迁移和侵袭数、N-cadherin蛋白表达量均显著降低（P<0.05），E-cadherin蛋白表达量显著增加（P<0.05）。过表达miR-503后SW579细胞活力、克隆形成数、迁移和侵袭数、N-cadherin蛋白表达量均显著降低（P<0.05），E-cadherin蛋白表达量显著增加（P<0.05）。miR-503是circ_0000064的靶基因，沉默circ_0000064可显著上调miR-503表达（P<0.05）。抑制miR-503表达可显著减弱沉默circ_0000064对SW579细胞生物学行为的影响（P<0.05）。结论 沉默circ_0000064可通过上调miR-503抑制甲状腺癌细胞增殖、迁移和侵袭，诱导细胞凋亡，从而抑制甲状腺癌进展。
Objective To investigate the effects of circular RNA 0000064 (circ_0000064) targeting miR-503 on the biological behavior of thyroid cancer cells.Methods The expression levels of circ_0000064 and miR-503 in the thyroid cancer tissues and the corresponding adjacent tissues of 21 cases were assessed using real-time quantitative PCR (RT-qPCR). Circ_0000064 small interfering RNA (si-circ_0000064), miR-503 mimic, si-circ_0000064 + miR-503 inhibitor (anti-miR-503) were transfected into thyroid cancer SW579 cells, respectively. The plate cloning formation test and cell counting kit (CCK-8) method were applied to calculate the proliferation of SW579 cells, and Transwell test was performed to detect the migration and invasion of SW579 cells. Flow cytometry was selected to detect the cell apoptosis, and Western blotting was employed to detect the expressions of epithelial cadherin (E-cadherin) and neural cadherin (N-cadherin) proteins. The targeted regulation relationship between circ_0000064 and miR-503 was confirmed by dual luciferase reporter assay and RT-qPCR.Results The expression of circ_0000064 was significantly higher in thyroid cancer tissues than in adjacent tissues (P<0.05), while the expression of miR-503 was significantly lower in thyroid cancer tissues than in adjacent tissues (P<0.05). The cell viability, colony formation numbers, migration and invasion numbers, and N-cadherin protein expression of SW579 cells were significantly decreased (P<0.05), while E-cadherin protein expression was significantly increased (P<0.05) after silencing circ_0000064. The cell viability, colony formation numbers, migration and invasion numbers, N-cadherin protein expression of SW579 cells were significantly decreased (P<0.05), while E-cadherin protein expression was significantly increased (P<0.05) after overexpressing miR-503. miR-503 was the target gene of circ_0000064, and silencing circ_0000064 significantly increased the expression of miR-503 (P<0.05). miR-503 inhibition significantly weakened the effects of silencing circ_0000064 on the biological behavior of SW579 cells (P<0.05).Conclusion Silencing circ_0000064 can inhibit the proliferation, migration and invasion, and induce the apoptosis of thyroid cancer cells by up-regulating the expression of miR-503, thereby inhibiting the progression of thyroid cancer.
龚烨,刘思吟,余喜梅. circ_0000064靶向miR-503对甲状腺癌细胞生物学行为的影响[J].肿瘤药学,2023,13(2):173-180 ( in Chinese)复制