miR-646 抑制骨肉瘤细胞的增殖和迁移的机制初探

Mechanism of Inhibition of Proliferation and Migration of Osteosarcoma Cells by miR-646

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。

Objective The mechanism of inhibition of the proliferation and migration of osteosarcoma cells by miR-646 was investigated. Meth- olts The effects of miR-646 on the proliferation of U2OS cells were detected by MTY assay. The effects of miR-646 on U2OS cell migration were analyzed by Transwell assay. MIRDB was used to predict the binding sites of miR-646 and epidermal growth factor receptor （EGFR）, which were confirmed in a luciferase reporter assay. Western blotting was conducted to detect the effect of miR-646 on the EGFR pathway. Results The MTF test showed that transfection of miR-646 sissfificanfly inhibited U2OS cell proliferation ; the 24 h inhibition rate was 21.76%±1.05%. In contrast, the 24 h promotion was 18.89%± 0.81%. Transwell experiments showed that miR-646 inhibited the migration of osteosarcoma U2OS cells. MIRDB predicted that miR-646 bound in the 3＇ -untranslated region of EGFR, and luciferase reporter gene experiments showed that miR-646 directly act- ed on EGFR. Western blotting experiments demonstrated that miR-646 inhibited the proliferation and migration of U2OS cells to some extent by in- hibiting the EGFR pathway. Conclusion miR-646 can inhibit the proliferation and migration of osteosarcoma cells by inhibiting the EGFR path- way.