山奈酚通过调控miR21抑制非小细胞肺癌A549细胞增殖的研究

Effect of Kaempferol on the Proliferation of Non-small Cell Lung Cancer A549 Cells by Regulating miR21

关键词:  山奈酚  miR21  非小细胞肺癌  增殖

Key words:  kaempferol  miR21  non-small cell lung cancer  proliferation

摘要:

目的 研究山奈酚（kaempferol，KA）对非小细胞肺癌A549细胞增殖的抑制作用及其作用机制。方法 运用CCK-8细胞活力检测法和集落形成实验研究不同浓度的KA对非小细胞肺癌A549细胞存活率的影响，确定给药浓度；将A549细胞分为对照组及KA各剂量组，采用qPCR检测不同浓度KA对A549细胞中miR21表达的影响；运用miR21 mimc转染A549细胞过表达miR21，采用qPCR和Western blotting检测miR21和不同浓度KA给药作用后对PTEN/AKT信号通路的影响。结果 CCK-8和集落形成实验证实KA可抑制非小细胞肺癌A549细胞的增殖；30，90μmol·L^-1 KA处理24 h均可降低A549细胞中miR21水平和p-AKT、c-Myc、Cyclin D1的表达水平，升高PTEN水平，而miR21 mimc转染后p-AKT、c-Myc、Cyclin D1的表达水平均升高，而PTEN水平降低，并经rescue实验证实miR21 mimc转染后可阻断KA抑制A549细胞增殖的作用。结论 KA能够通过调控miR21/PTEN/AKT信号通路抑制非小细胞肺癌A549细胞增殖。

Abstract:

OBJECTIVE To study the inhibitory effect of kaempferol(KA) on the proliferation of non-small cell lung cancer A549 cells and its mechanism. METHODS CCK-8 cell viability assay and colony formation assay were used to study the effect of KA on the survival rate of A549 cells and determine the treatment concentration of KA. A549 cells were divided into control group and different doses of KA group, and the effects of different concentrations of KA on the expression of microRNA in A549 cells were detected by qPCR. A549 cells were transfected by miR21 mimc. Overexpression of miR21 in A549 cells was detected by qPCR. The effects of miR21 and KA on PTEN/AKT signaling pathway were detected by Western blotting and qPCR. RESULTS CCK-8 and colony formation assays demonstrated that KA could inhibit the proliferation of non-small cell lung cancer A549 cells. KA treatment for 24 h at 30, 90 μmol·L^-1 obviously reduced the expression of miR21 and p-AKT, c-Myc, Cyclin D1 in A549 cells, and increased the expression of PTEN. However, the expression of p-AKT, c-Myc and Cyclin D1 was upregulated and PTEN was downregulated after miR21 mimc transfection. Finally the rescue experiment confirmed that KA induced inhibitory proliferation effect of A549 cells could be blocked by miR21 mimc transfection. CONCLUSION KA can inhibit the proliferation of non-small cell lung cancer A549 cells via regulating the miR21/PTEN/AKT signaling pathway.