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.
Ministry-of-Education Key Laboratory of Clinical Diagnostic Medicine, Laboratory Medical School, Chongqing Medical University, Chongqing 400016, China. *Corresponding author, E-mail:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2020 Nov;36(11):983-989.
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 Transwell 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.
目的 探讨miR-23b-3p对人宫颈癌CasKi细胞增殖、迁移及侵袭的影响。方法 体外培养人宫颈癌CasKi细胞和正常上皮HaCaT细胞。采用实时定量RT-PCR检测CasKi细胞和HaCaT细胞中miR-23b-3p的表达。通过脂质体法将合成的miR-23b-3p模拟物及其阴性对照转染至CasKi细胞。采用CCK-8法检测miR-23b-3p过表达对细胞增殖的影响。分别采用划痕愈合试验和Transwell试验观察CasKi细胞的迁移和侵袭能力。采用蛋白质印迹法检测N-钙黏蛋白、波形蛋白、E-钙黏蛋白、Snail、增殖细胞核抗原（PCNA）和细胞周期蛋白D1的蛋白表达。结果 CasKi细胞中miR-23b-3p的表达低于HaCaT细胞。与阴性对照组相比，转染miR-23b-3p模拟物后CasKi细胞中miR-23b-3p的表达显著上调。CCK-8法和蛋白质印迹法检测显示，用miR-23b-3p模拟物处理细胞后，细胞增殖受到抑制，PCNA和细胞周期蛋白D1的表达下调。同时，miR-23b-3p过表达后，CasKi细胞的迁移和侵袭能力显著受到抑制。此外，E-钙黏蛋白的表达上调，而波形蛋白、Snail和N-钙黏蛋白的表达水平显著下调。结论 miR-23b-3p过表达可能抑制人宫颈癌CasKi细胞的增殖、迁移、侵袭及上皮-间质转化过程。
