miR-646抑制骨肉瘤细胞的增殖和迁移的机制初探

发布时间：2018-03-27 15:26

本文选题：miR-　切入点：骨肉瘤　出处：《中国医科大学学报》2017年08期

【摘要】：目的探讨mi R-646对骨肉瘤细胞增殖和迁移的抑制作用。方法通过MTT实验检测mi R-646过表达或受到抑制后对U2OS细胞增殖的影响。Transwell实验分析mi R-646过表达或受到抑制后对U2OS细胞迁移的影响。MIRDB预测mi R-646与表皮生长因子受体(EGFR)的位点结合后,利用荧光素酶报告及实验检测mi R-646对EGFR的靶向作用,并通过Western blotting检测mi R-646对EGFR通路的作用。结果 MTT结果表明,转染mi R-646后U2OS细胞的增殖受到了显著地抑制,24 h时抑制率为21.76%±1.05%,相反,当mi R-646受到抑制后U2OS细胞的增殖得到了促进,24 h时促进的比率为18.89%±0.81%。Transwell实验发现mi R-646可以抑制骨肉瘤细胞系U2OS的迁移能力。随后MIRDB预测发现mi R-646与EGFR的3’UTR区域具有结合位点,荧光素酶报告基因实验证明了mi R-646可以直接作用于EGFR。Western blotting结果证明mi R-646对U2OS细胞增殖迁移能力的抑制在一定程度上是通过mi R-646对EGFR通路的抑制实现的。结论 mi R-646可以通过抑制EGFR通路的活性,进而抑制骨肉瘤细胞的增殖和迁移功能。
[Abstract]:Objective to investigate the inhibitory effect of mi R-646 on proliferation and migration of osteosarcoma cells. Methods MTT assay was used to detect the effect of mi R-646 overexpression or inhibition on the proliferation of U2OS cells. Transwell experiment was used to analyze the overexpression or inhibition of mi R-646 on U2OS cells. Effects of cell migration. MIRDB predicts that the site of mi R-646 binds to the epidermal growth factor receptor (EGFRR). Luciferase reports and experiments were used to detect the targeting effect of mi R-646 on EGFR and Western blotting to detect the effect of mi R-646 on EGFR pathway. After transfection of miR-646, the proliferation of U2OS cells was significantly inhibited by 21.76% 卤1.05% at 24 h. When the proliferation of U2OS cells was inhibited after MIR-646 was inhibited, the promotion rate of U2OS cells was 18.89% 卤0.81%.Transwell for 24 h. It was found that mi R-646 could inhibit the migration ability of osteosarcoma cell line U2OS. Subsequently, MIRDB predicted that mi R-646 had binding sites with 3'UTR region of EGFR. The experiment of luciferase reporter gene proved that mi R-646 could directly act on EGFR.Western blotting. The results showed that the inhibition of U2OS cell proliferation and migration by mi R-646 was to some extent realized by the inhibition of EGFR pathway by mi R-646. Conclusion\\\; By inhibiting the activity of the EGFR pathway, Furthermore, the proliferation and migration of osteosarcoma cells were inhibited.
【作者单位】：中国医科大学肿瘤医院辽宁省肿瘤医院骨与软组织肿瘤科;中国医科大学附属盛京医院骨科;沈阳医学院附属中心医院手外科;
【分类号】：R738.1

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