文章摘要
李莉,常春玲,魏海波.微小 RNA-466靶向调控 Twist1影响宫颈癌 SiHa细胞增殖和迁移[J].安徽医药,2022,26(1):156-160.
微小 RNA-466靶向调控 Twist1影响宫颈癌 SiHa细胞增殖和迁移
MicroRNA-466 targeted regulation of Twist1 on the proliferation and migration of cervical cancer SiHa cells
  
DOI:10.3969/j.issn.1009-6469.2022.01.036
中文关键词: 子宫肿瘤  碱性螺旋 -环-螺旋转录因子 1  miR-466  波形蛋白  迁移
英文关键词: Uterine neoplasms  Twist1  miR-466  Vimentin  Migration miRNA
基金项目:
作者单位
李莉 铁岭市中心医院妇产科辽宁铁岭 112000 
常春玲 铁岭市中心医院妇产科辽宁铁岭 112000 
魏海波 铁岭市中心医院妇产科辽宁铁岭 112000 
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中文摘要:
      目的研究微小 RNA-466(miR-466)靶向调控碱性螺旋 -环-螺旋转录因子 1(Twist1)对宫颈癌 SiHa细胞增殖和迁移的影响和机制。方法本研究起止时间为 2019年 1—10月。宫颈癌 Caski、SiHa、C33A细胞购自上海沪震生物科技有限公司;正常宫颈上皮 End1/E6E7细胞购自上海雅吉生物科技有限公司。定量即时聚合酶链锁反应( qRT-PCR)方法测定宫颈癌细胞中 miR-466表达变化。在宫颈癌 SiHa细胞中转染 miR-466模拟物( miR-466 mimics)噻唑蓝( MTT)方法检测细胞增殖变化, Transwell小室检测细胞迁移和侵袭能力变化,蛋白质印迹法( Western blotting)检测细,胞中波形蛋白( Vimentin)和上皮钙黏素( E-cadherin)蛋白表达变化。在线靶基因预测软件预测 miR-466的靶基因可能为 Twist1,荧光素酶报告系统鉴定靶向关系。将 Twist1过表达载体和 miR-466 mimics共转染到宫颈癌 SiHa细胞中,检测细胞增殖、迁移和侵袭能力变化。结果正常宫颈上皮 End1/E6E7细胞和宫颈癌 Caski、SiHa、C33A细胞中 miR-466表达水平依次为:(1.00±0.09)、(0.58±0.06)、(0.20±0.03)、
英文摘要:
      Objective To study the effect and mechanism of microRNA-466 (miR-466) targeted regulation of Twist basic helix-loop-helix transcription factor 1(Twist1) on the proliferation and migration of cervical cancer SiHa cells.Methods This study began in Janu-ary 2019 and ended in October 2019. Cervical cancer Caski, SiHa, and C33A cells were purchased from Shanghai Huzhen Biotechnolo-gy Co., Ltd.; normal cervical epithelial End1/E6E7 cells were purchased from Shanghai Yaji Biotechnology Co., Ltd. Quantitative realtime polymerase chain reaction (qRT PCR) was used to detect the expression of miR-466 in cervical cancer cells. miR-466 mimics was transfected into SiHa cells, methylthiazolyldiphenyl-tetrazolium bromide (MTT) method was used to detect cell proliferation, Transwellcells were used to detect cell migration and invasion, Western blotting was used to detect the expression of vimentin and epithelical cad-herin (E-cadherin). Online target gene prediction software predicted that the target gene of miR-466 might be Twist1, the target relation-ship was identified by luciferase report system. Twist1 overexpression vector and miR-466 mimics were co transfected into cervical can-cer SiHa cells to detect cell proliferation, migration and invasion.Results The expression levels of miR-466 in normal cervical epithe-lial End1/E6E7 cells and cervical cancer Caski, SiHa, and C33A cells were: (1.00±0.09), (0.58±0.06), (0.20±0.03), (0.45±0.04); the ex-pression level of miR-466 in the cancer cell lines Caski, SiHa, and C33A was lower than that in End1/E6E7 cells, and the differencewas statistically significant (P<0.0001). Compared with the miR-NC group, the cell A value of SiHa cells in the miR-466 group (0.29± 0.04 vs. 0.45±0.02) was decreased, the number of cell invasion [(81.46±6.35) vs. (117.95±11.62)] and the number of migration [(90.17± 7.43) vs. (147.15±12.04)] were decreased, Vimentin protein expression level was decreased, E-cadherin protein expression level was in-creased, the difference was statistically significant (P<0.001). Compared with the miR-466+pCDNA3.1 group, the A value of cervical cancer SiHa cells in the miR-466+pCDNA3.1-Twist1 group (0.41±0.04 vs. 0.30±0.03) was increased, and the number of cell migration [(127.34±10.26) vs. (93.05±6.84) and the number of invasion [(108.94±10.24) vs. (83.26±5.17)] were increased, Vimentin and Twist1 protein expression were increased, E-cadherin protein expression was decreased, the difference was statistically significant (P<0.05). Up-regulation of miR-466 targeted inhibition of Twist1 expression.Conclusion miR-466 targeted inhibition of Twist1 reduced the pro-liferation and migration of SiHa cells.
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