miR-183-5p对大肠癌细胞增殖、迁移能力的影响及机制

Effects of miR-183-5p on proliferation and migration of colorectal cancer cells

目的探讨miR-183-5p对大肠癌(CRC)细胞增殖、迁移能力的影响及机制。方法应用miRDB数据库,根据重组人抑制蛋白-2(PFN2)基因序列预测筛选的miR-183-5p;应用实时荧光定量PCR(qRT-PCR)法检测CRC组织及细胞系中miR-183-5p和PFN2 mRNA表达并分析其相关性;荧光素酶报告基因技术检测miR-183-5p与PFN23′非翻译区(3′UTR)特异性结合情况;应用qRT-PCR及Western blotting法检测转染miR-183-5p模拟物(mimics)后CRC细胞株PFN2表达情况;应用CCK8法及划痕实验检测转染miR-183-5p mimics后CRC细胞系的增殖、迁移能力。结果与正常大肠上皮细胞系及癌旁组织相比,PFN2在CRC细胞系及癌组织中的表达均下调(P均<0.05),miR-183-5p在CRC细胞系及癌组织中的表达均上调(P均<0.05),且二者表达呈负相关(r=-0.398,P=0.042);miR-183-5p可与PFN23′UTR特异性结合;转染miR-183-5p mimics后,CRC细胞系PFN2 mRNA及蛋白的表达受到抑制,CRC细胞增殖、迁移能力增强(P<0.05)。结论miR-183-5p可促进CRC细胞的增殖、迁移,其机制可能通过靶向下调PFN2表达实现。

Objective To investigate the effects of miR-183-5p on the proliferation and migration of colorectal cancer(CRC)cells.Methods The miRDB database was used to predict and screen out the upstream target miRNA(miR-183-5p)of PFN2;the expression levels of miR-183-5p and PFN2 mRNA in CRC tissues and cell lines were detected by qRT-PCR,and we analyzed the correlation between miR-183-5p and PFN2.Luciferase report gene technology was used to detect the specific binding of miR-183-5p to PFN23′untranslated region(3′UTR);after transfection of CRC cells,the expression of PFN2 was detected by real-time PCR(qRT-PCR)and Western blotting;the proliferation and migration abilities of CRC cells after transfection of miR-183-5p mimics were detected by CCK-8 and Scratch test.Results Compared with the normal colorectal epithelial cell line and paracancerous tissues,the expression of PFN2 was down-regulated,and miR-183-5p was up-regulated in CRC cells and cancer tissues(all P<0.05);the expression levels of them were negatively correlated(r=-0.398,P=0.042);miR-183-5p could specifically bind to PFN23′UTR.After transfection of miR-183-5p mimics,the expression of PFN2(mRNA and protein)in CRC cell lines was inhibited,and the proliferation and migration abilities increased(all P<0.05).Conclusion MiR-183-5p can promote the proliferation and migration of CRC cells probably by down-regulating PFN2 expression.