Gui Z L, Wu T L, Zhao G C, Lin Z X, Xu H G
Bratisl Lek Listy. 2017;118(8):449-452. doi: 10.4149/BLL_2017_087.
The aim of this study was to study the mechanism of miRNA-497 in the apoptosis of osteosarcoma cells.
MG-63 cells were divided into the three groups: NC, BL and miRNA groups, NC group were treated with nothing; BL group were transfected with blank vector; miRNA group were transfected with miRNA-497. Cell proliferation rate was detected by MTT method; Apoptosis rate was detected by flow cytometry and measuring the gene and protein expression of MAPK, Erk and P 21 by RT-PCR and Western blot.
The cell proliferation rate of miRNA group was significantly lower compared to NC group and BL group (p < 0.05); while the apoptosis rate of miRNA group (32.17 ± 3.23 %) was significantly higher than that of NC group (8.40 ± 1.78 %) and BL group (8.83 ± 0.99 %) (p < 0.05, respectively). Regarding the gene expression detection, we found that gene and protein expressions of MAPK, Erk and P21 of miRNA group were significantly different compared to NC and BL groups (p < 0.05, respectively).
MiR-497 can activate P21 expression by inhibiting the expression of MAPK/Erk signaling pathway, thus promoting the apoptosis of osteosarcoma cells (Fig. 5, Ref. 18).
本研究旨在探讨miRNA - 497在骨肉瘤细胞凋亡中的作用机制。
将MG - 63细胞分为三组:NC组、BL组和miRNA组。NC组不做处理;BL组转染空载体;miRNA组转染miRNA - 497。采用MTT法检测细胞增殖率;通过流式细胞术检测凋亡率,并采用RT - PCR和蛋白质印迹法检测MAPK、Erk和P21的基因及蛋白表达。
miRNA组细胞增殖率显著低于NC组和BL组(p < 0.05);而miRNA组的凋亡率(32.17 ± 3.23%)显著高于NC组(8.40 ± 1.78%)和BL组(8.83 ± 0.99%)(p均 < 0.05)。在基因表达检测方面,我们发现miRNA组中MAPK、Erk和P21的基因及蛋白表达与NC组和BL组相比有显著差异(p均 < 0.05)。
MiR - 497可通过抑制MAPK/Erk信号通路的表达来激活P21表达,从而促进骨肉瘤细胞凋亡(图5,参考文献18)。