马钱苷联合miR-3619-5p靶向迁移侵袭增强因子1对宫颈癌SiHa细胞迁移和凋亡的影响

目的 研究马钱苷联合miR-3619-5p靶向迁移侵袭增强因子1(migration and invasion enhancer 1,MIEN1)对宫颈癌SiHa细胞迁移和凋亡的影响。方法 采用qRT-PCR法检测宫颈癌SiHa、Hela、CasKi细胞和正常宫颈上皮Ect1/E6E7细胞中miR-3619-5p m RNA表达。在宫颈癌SiHa细胞中转染miR-3619-5p mimics上调miR-3619-5p的表达，给予马钱苷处理，采用CCK-8法分析细胞增殖；流式细胞术分析细胞凋亡；Transwell小室分析细胞迁移和侵袭能力的变化；Western blotting法分析剪切型半胱氨酸天冬氨酸蛋白酶-3(cleaved cystein-asparate protease-3,cleaved Caspase-3)和基质金属蛋白酶-2(matrix metalloprotease-2,MMP-2)蛋白表达。生物信息学软件预测miR-3619-5p的靶基因，利用荧光素酶报告系统鉴定二者的靶向关系。在宫颈癌SiHa细胞中共转染miR-3619-5p mimics和MIEN1过表达载体...

Effect of loganin combined with miR-3619-5p targeting MIEN1 on migration and apoptosis of cervical cancer SiHa cells

Objective To investigate the effect of loganin combined with miR-3619-5p targeting migration and invasion enhancer 1 (MIEN1) on migration and apoptosis of cervical cancer SiHa cells. Methods miR-3619-5p mRNA expression in cervical cancer SiHa, Hela, CasKi cells and normal cervical epithelial Ect1/E6E7 cells were detected by qRT-PCR. Cervical cancer SiHa cells were transfected with miR-3619-5p mimics to up-regulate the expression of miR-3619-5p and treated with loganine. Cell proliferation was analyzed by CCK-8 method; Cell apoptosis was analyzed by flow cytometry; Migration and invasion abilities of cells were analyzed by Transwell chamber; Cleaved cystein-asparate protease-3 (cleaved Caspase-3) and matrix metalloprotease-2 (MMP-2) protein expressions were detected by Western blotting. Bioinformatics software was used to predict the target genes of miR-3619-5p, and luciferase reporter system was used to identify the targeting relationship between the two. Cervical cancer SiHa cells were co-transfected with miR-3619-5p mimics and MIEN1 overexpression vector to investigate the changes in cell proliferation, apoptosis, migration and invasion. Results miR-3619-5p mRNA expression level in cervical cancer SiHa, Hela and CasKi cells was lower than that in normal cervical epithelial Ect1/E6E7 cells (P < 0.05), and miR-3619-5p expression level in cervical cancer SiHa cells was the lowest. After up-regulation of miR-3619-5p, given loganin or a combination of two treatments, cells proliferation, migration and invasion abilities were decreased (P < 0.05), cell apoptosis was increased (P < 0.05), cleaved Caspase-3 protein expression was increased (P < 0.05), and MMP-2 protein expression was decreased (P < 0.05). And combined treatment of two had a stronger effect on cells proliferation, apoptosis, invasion and migration. miR-3619-5p targeted MIEN1 expression. MIEN1 overexpression vector reversed the effects of loganin combined with miR-3619-5p on proliferation, apoptosis, migration and invasion of cervical cancer SiHa cells (P < 0.05). Conclusion Loganin combined with miR-3619-5p targeting MIEN1 can inhibit the proliferation, migration and invasion of cervical cancer SiHa cells, and promote cell apoptosis.