Glioma is the most common type of primary brain tumor and has an undesirable prognosis. Autophagy plays an important role in cancer therapy, but it is effect is still not definite. P53 is an important tumor suppressor gene and protein that is closely to autophagy. Our aim was to study the effect of β-asarone on inhibiting cell proliferation in human glioma U251 cells and to detect the effect of the inhibition on autophagy through the P53 signal pathway. For cell growth, the cells were divided into four groups: the model, β-asarone, temozolomide (TMZ), and co-administration groups. For cell autoghapy and the P53 pathway, the cells were divided into six groups: the model, β-asarone, 3MA, Rapa, Pifithrin-µ, and NSC groups. The counting Kit-8 assay and flow cytometry (FCM) were then used to measure the cell proliferation and cycle. Electron microscopy was used to observe autophagosome formation. Cell immunohistochemistry/-immunofluorescence, FCM and Western blot (WB) were used to examine the expression of Beclin-1 and P53. The levels of P53 and GAPDH mRNA were detected by RT-PCR. Using WB, we determined autophagy-related proteins Beclin-1, LC3-II/I, and P62 and those of the P53 pathway-related proteins P53, Bcl-2, mTOR, P-mTOR, AMPK, P-AMPK, and GAPDH. We got the results that β-asarone changed the cellular morphology, inhibited cell proliferation, and enhanced the expression of P53, LC3-II/I, Beclin-1, AMPK, and pAMPK while inhibiting the expression of P62, Bcl-2, mTOR, and pmTOR. All the data suggested that β-asarone could reduce the cell proliferation and promote autophagy possible via the P53 pathway in U251 cells.

译文

神经胶质瘤是最常见的原发性脑肿瘤类型,预后不良。自噬在肿瘤治疗中起着重要作用,但其作用尚不明确。P53是一种重要的抑癌基因和蛋白,与自噬密切。我们的目的是研究 β-asarone对人神经胶质瘤U251细胞增殖的抑制作用,并通过P53信号途径检测其对自噬的抑制作用。对于细胞生长,将细胞分为四组: 模型,β-细辛醚,替莫唑胺 (TMZ) 和共同给药组。对于细胞自体和P53途径,将细胞分为六组: 模型,β-细辛醚,3MA,Rapa,Pifithrin-µ 和NSC组。然后使用计数试剂盒-8分析和流式细胞术 (FCM) 测量细胞增殖和周期。使用电子显微镜观察自噬体的形成。细胞免疫组织化学/免疫荧光,FCM和Western blot (WB) 检测Beclin-1和p53的表达。Rt-pcr检测P53和GAPDH mRNA水平。使用WB,我们确定了自噬相关蛋白Beclin-1,LC3-II/I和P62以及P53途径相关蛋白P53,Bcl-2,mTOR,P-mTOR,AMPK,P-AMPK和GAPDH的自噬相关蛋白。结果表明,β-asarone改变了细胞形态,抑制了细胞增殖,增强了P53,LC3-II/I,Beclin-1,AMPK和pAMPK的表达,同时抑制了P62,Bcl-2,mTOR和pmTOR的表达。所有数据表明,β-asarone可能通过P53途径降低U251细胞的增殖并促进自噬。

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