过表达Hsa-miR-23b-3p抑制CasKi人宫颈癌细胞的增殖、侵袭和迁移及上皮间质转化  

Over-expression of Hsa-miR-23b-3p suppresses proliferation,migration,invasion and epithelial-mesenchymal transition of human cervical cancer CasK i cells

作　　者：胡静 孙子久 胡凯 唐敏[1] 孙恃雷 方玉婷 余伙梅 张彦[1] HU Jing;SUN Zijiu;HU Kai;TANG Min;SUN Shilei;FANG Yuting;YU Huomei;ZHANG Yan(Ministry-of-Education Key Laboratory of Clinical Diagnostic Medicine,Laboratory Medical School,Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016

出　　处：《细胞与分子免疫学杂志》2020年第11期983-989,共7页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81974449)。

摘　　要：目的探讨miR-23b-3p对CasKi人宫颈癌细胞增殖、侵袭与迁移的影响。方法体外培养CasKi人宫颈癌细胞和HaCaT正常上皮细胞,采用实时定量PCR检测两种细胞miR-23b-3p的表达水平。采用脂质体转染miR-23b-3p模拟物(miR-23b-3p mimic)的方法建立外源性miR-23b-3p过表达的CasKi细胞,阴性对照组中转染无关序列;CCK-8法检测过表达miR-23b-3p后CasKi细胞的增殖,TranswellTM侵袭实验检测细胞的侵袭能力、TranswellTM迁移实验和划痕愈合实验检测CasKi细胞的迁移能力;Western blot法检测神经钙黏蛋白(N-cadherin)、波形蛋白(vimentin)、上皮钙黏蛋白(E-cadherin)、Snail、增殖细胞核抗原(PCNA)、细胞周期蛋白D1(cyclin D1)的蛋白表达。结果与HaCaT细胞相比,CasKi细胞miR-23b-3p表达水平较低。与阴性对照组相比,转染miR-23b-3p mimic的CasKi细胞miR-23b-3p水平明显上调;miR-23b-3p表达上调后,细胞增殖能力减弱,PCNA及cyclin D1表达减少;同时,过表达miR-23b-3p后,CasKi细胞的侵袭及迁移能力均明显受抑制;同时增加E-cadherin蛋白的表达,而抑制vimentin、Snail和N-cadherin蛋白的表达。结论过表达miR-23b-3p抑制CasKi宫颈癌细胞的增殖、侵袭、迁移和上皮间质转化(EMT)。Objective To investigate the effects of miR-23b-3p on proliferation,migration and invasion of human cervical carcinoma CasKi cells.Methods Human cervical carcinoma CasKi cells and normal epithelial HaCaT cells were cultured in vitro.Real-time quantitative RT-PCR was conducted to detect the expression of miR-23b-3p in CasKi and HaCaT cells.Synthetic miR-23b-3p mimic and its negative control were transfected into CasKi cells by liposome method.The effects of miR-23b-3p over-expression on cell proliferation were detected by CCK-8 assay.Wound scratch healing assay and TranswellTM assay were used to observe the migration and invasion abilities of CasKi cells,respectively.Western blot analysis was used to detect the protein expression of N-cadherin,vimentin,E-cadherin,Snail,PCNA and cyclin D1.Results The expression of miR-23b-3p in CasKi cells was lower than that of HaCaT cells.Compared with the negative control group,the expression of miR-23b-3p were significantly up-regulated in CasKi cells after transfected with miR-23b-3p mimic.CCK-8 and Western blot assays showed that the proliferation was inhibited and the expression of PCNA and cyclin D1 were down-regulated after the cells were treated with miR-23b-3p mimic.At the same time,after over-expression of miR-23b-3p,the migration and invasion abilities of the CasKi cells were significantly inhibited.In addition,the expression of E-cadherin was up-regulated,while vimentin,Snail and N-cadherin expression levels were significantly down-regulated.Conclusion Over-expression of miR-23b-3p may suppress the proliferation,migration,invasion and epithelial-mesenchymal transition process of human cervical cancer CasKi cells.

关 键 词：宫颈癌 miR-23b-3p 细胞增殖 细胞侵袭 细胞迁移 

分 类 号：Q279[生物学—细胞生物学] R737.33[医药卫生—肿瘤]