摘要
目的:阐明下调miR-374a靶向转录因子21(TCF21)调控宫颈癌细胞增殖、迁移、侵袭的分子机制。方法:采用Realtime PCR法检测微小RNA(miR)-374a在正常宫颈上皮细胞和宫颈癌细胞中的表达变化。在宫颈癌细胞Si Ha中转染miR-374a inhibitor,MTT检测细胞增殖,Transwell小室检测细胞侵袭以及迁移能力变化,Western blot检测细胞中基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)蛋白表达变化。在线靶基因预测软件预测miR-374a的靶基因可能为TCF21,荧光素酶报告系统鉴定靶向关系。在宫颈癌细胞Si Ha中共转染miR-374a inhibitor和TCF21 siRNA,检测细胞增殖、侵袭、迁移变化。结果:miR-374a在正常宫颈上皮细胞中的表达水平明显低于宫颈癌细胞。转染miR-374a inhibitor后的宫颈癌细胞Si Ha增殖、迁移和侵袭能力下降,细胞中MMP-2和MMP-9蛋白表达减少。miR-374a靶向负调控TCF21表达。TCF21 siRNA逆转miR-374a inhibitor对宫颈癌细胞Si Ha增殖、侵袭和迁移的抑制作用。结论:下调miR-374a通过靶向TCF21抑制宫颈癌细胞Si Ha增殖、迁移、侵袭。
Objective: To elucidate the molecular mechanism of down-regulation of miR-374a through targeting TCF21 to inhibit the proliferation,migration and invasion of cervical cancer cells. Methods: Realtime PCR was used to detect the expression of miR-374a in normal cervical epithelial cells and cervical cancer cells. miR-374a Inhibitor was transfected into cervical cancer cell line Si Ha.MTT was used to detect the proliferation. Transwell cells were used to detect the invasion and migration. Western blot was used to detect the expressions of MMP-2 and MMP-9. On-line target gene prediction software predicted that the target gene of miR-374a may be TCF21. Luciferase reporting system identified the target relationship. miR-374a Inhibitor and TCF21 siRNA were co-transfected into cervical cancer cell Si Ha to detect the cell proliferation,invasion and migration. Results: The expression of miR-374a in the normal cervical epithelial cells was significantly lower than that in cervical cancer cells,the proliferation,migration and invasion of Si Ha cells transfected with miR-374a inhibitor decreased,and the expressions of MMP-2 and MMP-9 decreased in cells.The expression of TCF21 was negatively regulated by miR-374a. TCF21 siRNA reversed the inhibitory effects of miR-374a inhibitor on the proliferation,invasion and migration of Si Ha cells. Conclusion: Down-regulation of miR-374a inhibits the proliferation,migration and invasion of Si Ha cells by targeting TCF21.
作者
向雪芹
崔权哲
詹文斌
陆静
Xiang Xueqin;Cui Quanzhe;Zhan Wenbin;Lu Jing(Department of Gynaecology and Obstetrics,Chengdu Third People's Hospital,Chengdu 610014,China;Department of Pathology,Chengdu Third People's Hospital,Chengdu 610014,China)
出处
《中国药师》
CAS
2021年第4期630-635,共6页
China Pharmacist
基金
四川省卫生和计划委员会科研项目(编号:16PJ046)。
