摘要
目的:探索环状RNA(Circular RNA,circRNA)circ_007142对甲状腺癌细胞增殖、迁移和凋亡的影响及其机制。方法:应用实时定量PCR(qRT-PCR)测定甲状腺癌组织中circ_007142和miR-874表达。按si-NC组、小干扰RNA(Small interference RNA,siRNA) circ_007142(si-circ_007142)组、miR-NC组、miR-874组、si-circ_007142+anti-miR-NC组、si-circ_007142+anti-miR-874组的分组在甲状腺癌TPC-1细胞中转染si-NC、si-circ_007142、miR-NC、miR-874 mimics、anti-miR-NC、anti-miR-874。分别采用细胞计数试剂盒8(CCK-8)、集落形成实验、划痕试验、流式细胞术和免疫印迹实验(Western blot)检测细胞活力、集落形成数、迁移、细胞凋亡、上皮型钙黏蛋白(E-cadherin)和神经型钙黏蛋白(N-cadherin)表达。荧光素酶活性实验分析circ_007142和miR-874靶向关系。结果:甲状腺癌组织中circ_007142表达量显著高于癌旁组织,miR-874表达量显著低于癌旁组织(P<0.05)。si-circ_007142组TPC-1细胞中miR-874表达量、凋亡率、E-cadherin蛋白表达量高于si-NC组,且si-circ_007142组细胞活力、集落形成数、划痕愈合率、N-cadherin蛋白表达量低于si-NC组(P<0.05或P<0.01)。miR-874组TPC-1细胞的活力、集落形成数、划痕愈合率、N-cadherin蛋白表达量均比miR-NC组低,凋亡率、E-cadherin蛋白表达量比miR-NC组高(P<0.05或P<0.01)。circ_007142可以靶向调控miR-874的表达。si-circ_007142+anti-miR-874组TPC-1细胞的miR-874表达量、凋亡率、E-cadherin蛋白表达量低于si-circ_007142+anti-miR-NC组,细胞活力、集落形成数、划痕愈合率、N-cadherin蛋白表达量高于si-circ_007142+anti-miR-NC组(P<0.05或P<0.01)。结论:干扰circ_007142通过靶向miR-874,抑制甲状腺癌细胞的增殖、迁移,以及促进凋亡。
Objective: To explore the effects of circular RNA(circRNA) circ_0007142 on the proliferation, migration and apoptosis of thyroid cancer cells and its mechanism.Methods: Real-time quantitative PCR(qRT-PCR) was used to determine the expressions of circ_0007142 and miR-874 in thyroid cancer tissues. According to the grouping of si-NC group, small interference RNA(siRNA) circ_0007142(si-circ_0007142) group, miR-NC group, miR-874 group, si-circ_0007142+anti-miR-NC group and si-circ_0007142+anti-miR-874 group, the thyroid cancer TPC-1 cells were respectively transfected with si-NC, si-circ_0007142, miR-NC, miR-874 mimics, anti-miR-NC and anti-miR-874. Cell viability, number of colonies formed, migration, apoptosis, epithelial cadherin(E-cadherin) and neuronal cadherin(N-cadherin) expressions were determined respectively by cell counting kit 8(CCK-8), colony formation test, scratch test, flow cytometry and Western blot. Luciferase activity experiment analyzed the targeting relationship between circ_0007142 and miR-874. Results: The expression of circ_0007142 in thyroid cancer tissue was higher than that in adjacent tissues, and the expression of miR-874 was lower than that in adjacent tissues(P<0.05). The expression of miR-874, apoptosis rate and E-cadherin protein expression in TPC-1 cells of si-circ_0007142 group were higher than those in si-NC group, and the cell viability, colony formation number, scratch healing rate and N-cadherin protein expression were lower than those of si-NC group(P<0.05 or P<0.01). TPC-1 cell viability, colony formation number, scratch healing rate and N-cadherin protein expression in miR-874 group were lower than those in miR-NC group, and the apoptosis rate and E-cadherin protein expression were higher than those in miR-NC group(P<0.05 or P<0.01). circ_0007142 could target and regulate the expression of miR-874. The miR-874 expression, apoptosis rate and E-cadherin protein expression of TPC-1 cells in si-circ_0007142+anti-miR-874 group were lower than those in si-circ_0007142+anti-miR-NC g
作者
黄寅
蒋智军
徐卫红
Huang Yin;Jiang Zhijun;Xu Weihong(Department of General Surgery,Jinhua Hospital of Traditional Chinese Medicine,Zhe-jiang Jinhua 321017,China;Department of Surgery,First Affiliated Hospital of Zhejiang University)
出处
《中国药师》
CAS
2021年第12期2176-2181,共6页
China Pharmacist
基金
金华市科技计划项目(编号:2010A33254)。
