Pae下调miR-106a-5p/PTEN对白血病细胞增殖、凋亡的影响

The effect of paeonol on the proliferation and apoptosis of leukemia cells by down-regulating miR-106a-5p/PTEN

为探究丹皮酚(paeonol,Pae)调控miR-106a-5p/第10号染色体缺失的磷酸酶及张力蛋白同源物(phosphatase and tension homologue deleted on chromosome 10,PTEN)通路对白血病细胞增殖和凋亡的影响,采用双荧光素酶报告基因分析法验证miR-106a-5p对PTEN的靶向作用。用不同浓度Pae处理K562细胞,分别构建miR-106a-5p抑制表达和经Pae处理的miR-106a-5p过表达或PTEN抑制表达K562细胞株,qRT-PCR检测miR-106a-5p和PTEN的mRNA表达水平;Western blotting检测PTEN、细胞周期素D1(CyclinD1)、Bcl-2、p21和Bax蛋白表达水平;MTT法检测K562细胞增殖能力;FACS检测细胞凋亡率。结果显示,PTEN是miR-106a-5p的靶基因,miR-106a-5p负向调控PTEN的表达;Pae处理可抑制K562细胞的增殖,促进其凋亡,并可抑制白血病细胞miR-106a-5p、CyclinD1和Bcl-2的表达,促进PTEN、Bax和p21的表达(均P<0.05);同时,抑制miR-106a-5p表达可抑制K562细胞增殖,促进其凋亡(均P<0.05);miR-106a-5p过表达或抑制PTEN表达均可部分逆转Pae对K562细胞增殖凋亡的影响(有P<0.05)。由此,Pae通过下调miR-106a-5p/PTEN信号通路蛋白抑制白血病细胞增殖,促进其凋亡。

To investigate the effect of paeonol(Pae) on the proliferation and apoptosis of leukemia cells by regulating miR-106 a-5 p/phosphatase and tension homologue deleted on chromosome 10(PTEN), dual luciferase reporter assay was used to verify the targeted relationship between miR-106 a-5 p and PTEN. The K562 cells were treated with different concentrations of Pae, as well as the K562 cell lines with inhibition of miR-106 a-5 p, and the K562 cell lines with overexpression of miR-106 a-5 p or inhibition of PTEN. The expression of miR-106 a-5 p and PTEN mRNA were detected by qRT-PCR, the expression of PTEN, CyclinD1, Bcl-2, p21 and Bax proteins were detected by Western blotting, the cell proliferation was detected by MTT assay, and the cell apoptosis rate was detected by FACS. The results showed that PTEN was the target gene of miR-106 a-5 p, and miR-106 a-5 p negatively regulated the expression of PTEN. Pae treatment inhibited the proliferation of K562 cells, promoted its apoptosis, inhibited the expression of miR-106 a-5 p, CyclinD1 and Bcl-2, and promoted the expression of PTEN, Bax and p21(all P<0.05). The inhibition of miR-106 a-5 p inhibited the proliferation and promoted apoptosis of K562 cells(all P<0.05). The effect of Pae on proliferation and apoptosis of K562 cells was partially reversed by the overexpression of miR-106 a-5 p or the inhibition of PTEN(P<0.05). In conclusion, the study demonstrates that Pae could inhibit the proliferation and promote the apoptosis of leukemia cells by downregulating miR-106 a-5 p/PTEN signaling pathway.