人参皂苷 Rh2 通过靶向 microRNA-128 抑制神经胶质瘤细胞增殖。
Ginsenoside Rh2 inhibits glioma cell proliferation by targeting microRNA-128.
机构信息
Department of Neurosurgery, Southwest Hospital, Third Military Medical University, Chongqing, China.
出版信息
Acta Pharmacol Sin. 2011 Mar;32(3):345-53. doi: 10.1038/aps.2010.220.
AIM
To examine the influence of ginsenoside Rh2 (Rh2), a triterpene saponin extracted from the traditional medicinal plant ginseng, on the expression of miRNAs in human glioma cells.
METHODS
The expression profile of miRNA (miR) was analyzed in human U251, T98MG and A172 glioma cells using a miRNA array and quantitative real-time PCR. Cell viability was assessed using a colorimetric assay (cell counting kit-8). Transfection of miR-128 was performed using Lipofectamine 2000. Caspase 3 activity was determined using a caspase colorimetric assay kit. Apoptosis was assessed using annexin V and propidium iodide staining. Protein expression was determined with Western blot analysis. miRNA-128 targeting activity was measured using a luciferase reporter assay.
RESULTS
In U251 cells treated with Rh2 (12 μg/mL), 14 of 452 human miRNAs were up-regulated and 12 were down-regulated as detected with the miRNA array assay. The up-regulation of miR-128 by Rh2 was further verified in human U251, T98MG and A172 cells using quantitative real-time PCR. In U251 cells, transfection of a miR-128 inhibitor (50 nmol/L) prevented the overexpression of miR-128 by Rh2, and significantly blunted Rh2-induced cytotoxicity, apoptosis, caspase 3 activation, transcriptional activation of E2F3a, a miR-128 target gene, as well as E2F3a protein expression.
CONCLUSION
The anti-proliferative effect of Rh2 in human glioma cells was mediated in part through up-regulation of miRNA-128 expression.
目的
研究从传统药用植物人参中提取的三萜皂苷人参皂苷 Rh2 对人胶质瘤细胞中 miRNA 表达的影响。
方法
采用 miRNA 芯片和实时定量 PCR 分析人 U251、T98MG 和 A172 神经胶质瘤细胞中 miRNA（miR）的表达谱。采用比色法（细胞计数试剂盒-8）评估细胞活力。使用 Lipofectamine 2000 转染 miR-128。使用 caspase 比色测定试剂盒测定 caspase 3 活性。用 Annexin V 和碘化丙啶染色法评估细胞凋亡。用 Western blot 分析检测蛋白表达。采用荧光素酶报告基因检测法测定 miRNA-128 的靶向活性。
结果
在 Rh2（12 μg/ml）处理的 U251 细胞中，通过 miRNA 芯片检测到 452 个人类 miRNA 中有 14 个上调，12 个下调。用实时定量 PCR 进一步验证了 Rh2 对人 U251、T98MG 和 A172 细胞中 miR-128 的上调。在 U251 细胞中，转染 miR-128 抑制剂（50 nmol/L）可阻止 Rh2 引起的 miR-128 过表达，并显著减弱 Rh2 诱导的细胞毒性、细胞凋亡、caspase 3 激活、miR-128 靶基因 E2F3a 的转录激活以及 E2F3a 蛋白表达。
结论
Rh2 对人神经胶质瘤细胞的抗增殖作用部分是通过上调 miR-128 表达介导的。
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