下调胰岛素样生长因子抑制miR-767-5p的表达对乳腺癌细胞增殖、迁移、侵袭及上皮间充质转化的影响
收稿日期: 2023-08-09
网络出版日期: 2023-12-11
基金资助
海南省自然科学基金项目(823MS143)
Suppression of miR-767-5p expression of inhibits breast cancer cell proliferation， migration， invasion and EMT through down-regulating IGF1
Received date: 2023-08-09
Online published: 2023-12-11
目的 研究抑制miR-767-5p对乳腺癌细胞增殖、迁移、侵袭和上皮间充质转化（EMT）的影响。 方法 体外培养乳腺癌细胞，将miR-767-5p inhibitor和inhibitor-NC转染到乳腺癌细胞中。细胞分为3组：空白对照组（Control组），miR-767-5p inhibitor组和inhibitor-NC组，采用CCK-8法、Transwell法、划痕实验检测miR-767-5p抑制剂对乳腺癌细胞增殖、侵袭和迁移能力的影响。Western blot检测迁移相关蛋白基质金属蛋白酶-2（MMP-2）和MMP-9的表达水平。双荧光素酶报告基因实验验证miR-767-5p和胰岛素样生长因子-1（IGF1）的靶向结合作用。RT-qPCR和Western blot实验检测IGF1 mRNA和蛋白的表达水平。采用Western blot检测EMT相关蛋白的表达水平。 结果 CCK-8实验结果显示，与Control组和inhibitor-NC组相比，miR-767-5p inhibitor组显著抑制MCF-7细胞的增殖（P < 0.05）。Transwell实验结果表明，与Control组和inhibitor-NC组相比，miR-767-5p inhibitor组乳腺癌细胞侵袭细胞数显著下降（P < 0.05）。划痕实验结果显示，与Control组和inhibitor-NC组相比，miR-767-5p inhibitor组乳腺癌细胞迁移率显著下降（P < 0.05）。荧光素酶报告基因实验结果显示，miR-767-5p能够靶向结合IGF1。Western blot结果表明，与Control组和inhibitor-NC组相比，miR-767-5p inhibitor组的E-cadherin蛋白表达水平显著增高（P < 0.05），N-cadherin、MMP-2和MMP-9蛋白的表达水平显著降低（P < 0.05）。与Control组和inhibitor-NC组相比，miR-767-5p inhibitor组中IGF1的mRNA和蛋白表达水平显著降低（P < 0.05）。 结论 抑制miR-767-5p的表达可通过下调IGF1抑制乳腺癌细胞的增殖、侵袭、迁移及EMT的发生。
关键词： 乳腺癌; miR-767-5p; 胰岛素样生长因子-1; 上皮间充质转化; 迁移侵袭
刘海英 , 陈峰 , 姚嘉 . 下调胰岛素样生长因子抑制miR-767-5p的表达对乳腺癌细胞增殖、迁移、侵袭及上皮间充质转化的影响[J]. 实用医学杂志, 2023 , 39(22) : 2878 -2884 . DOI: 10.3969/j.issn.1006-5725.2023.22.004
Objective To investigate the effect of inhibiting miR-767-5p on migration， invasion and epithelial-mesenchymal transition（EMT） of breast cancer cells. Methods Breast cancer cells were cultured in vitro， and miR-767-5p inhibitor and inhibitor NC were transfected into breast cancer cells. The cells were divided into three groups： Control group， miR-767-5p inhibitor group and inhibitor-NC group. The effects of miR-767-5p inhibitor on the proliferation， invasion and migration of breast cancer cells were detected by CCK-8 assay， Transwell Migration assay and scratch test. Western blot was used to detect the expression levels of migration related proteins MMP-2 and MMP-9. The dual luciferase reporter gene experiment verifies the targeted binding effect of miR-767-5p and IGF1. RT-qPCR and Western blot experiments were used to detect the expression levels of IGF1 mRNA and protein. Western blot was used to detect the expression level of EMT related proteins. Results CCK-8 assay showed that compared with the control group and inhibitor-NC group， the miR-767-5p inhibitor group significantly inhibited the proliferation of MCF-7 cells（P < 0.05）. Transwell assay showed that the number of invasive cells of breast cancer cells in miR-767-5p inhibitor group was significantly lower than that in control group and inhibitor NC group （P < 0.05）. Scratch test showed that compared with the control group and inhibitor NC group， the migration rate of breast cancer cells in miR-767-5p inhibitor group decreased significantly （P < 0.05）. Luciferase reporter gene assay showed that miR-767-5p targeted IGF1 binding. Western blot showed that compared with the control group and inhibitor NC group， the expression level of E-cadherin protein in the miR-767-5p inhibitor group was significantly increased （P < 0.05）， while the expression levels of N-cadherin， MMP-2， and MMP-9 protein were significantly reduced （P < 0.05）. Compared with the control group and inhibitor-NC group， the mRNA and protein expression levels of IGF1 in the miR-767-5p inhibitor group were significantly reduced （P < 0.05）. Conclusion Suppressing the expression of miR-767-5p may inhibit the proliferation， invasion， migration and EMT of breast cancer cells by down-regulating IGF1.
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