The effect of Ganoderic Acid A on miR-17-5p and miR-181b expression level and apoptosis induction in human leukemia Nalm-6 cells

Faezeh Mortazavie, Simin Taheri, Parisa Tandel, F. Zare, Gholmhossein Tamaddon
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引用次数: 1

Abstract

Background: In various cancers, Ganoderic Acid A (GAA), an active triterpenoid derived from Ganoderma lucidum, has been proved to show potent anti-tumor effects. However, the possible impacts of GAA on the human leukemia cell line (Nalm-6) are not fully elucidated. Therefore, this research aimed to study the antineoplastic effect of GAA on Nalm-6 cells. Materials and Methods: In this laboratory trial study, Nalm6 cells were cultured in vitro and treated with different doses of GAA (25, 50, 100, 200, and 400 μg/mL) for 24, 48, and 72 hours. The optimal treatment concentration of GAA was determined by the MTT assay. Flow cytometry was used to determine the death of Nalm-6 cells caused by GAA treatment by utilizing FITC-conjugated propidium iodide (PI) and annexin V staining. After incubation, the expression levels of miR-17-5p and miR-181b were monitored using real-time polymerase chain reaction (PCR). Results: Based on the half-maximal inhibitory concentration (IC50) measurements of the MTT assay, the optimal treatment concentration of GAA was 140 μg/mL (in a dose and time-dependent manner, p<0.0001). The GAA treatment was selectively toxic to the leukemia Nalm-6 cells and could remarkably induce cell apoptosis (p<0.0001). Besides, GAA downregulated the expression of miR-17-5p and miR-181b in the Nalm-6 cells compared with the untreated cells (P=0.0067 and P=0.0014, respectively). Conclusions: Based on the present findings, GAA merits further investigation as a promising natural reagent for treating hematologic malignancies.
灵芝酸A对人白血病Nalm-6细胞中miR-17-5p、miR-181b表达水平及诱导凋亡的影响
背景:从灵芝中提取的活性三萜灵芝酸A (GAA)已被证明在多种癌症中具有有效的抗肿瘤作用。然而,GAA对人白血病细胞系(Nalm-6)可能产生的影响尚不完全清楚。因此,本研究旨在研究GAA对Nalm-6细胞的抗肿瘤作用。材料与方法:体外培养Nalm6细胞,分别用不同剂量的GAA(25、50、100、200、400 μg/mL)处理24、48、72小时。采用MTT法确定GAA的最佳处理浓度。流式细胞术采用fitc偶联碘化丙啶(PI)和膜联蛋白V染色检测GAA处理后Nalm-6细胞的死亡情况。孵育后,采用实时聚合酶链反应(PCR)监测miR-17-5p和miR-181b的表达水平。结果:根据MTT法的半最大抑制浓度(IC50)测定,GAA的最佳处理浓度为140 μg/mL(呈剂量和时间依赖性,p<0.0001)。GAA对白血病Nalm-6细胞有选择性毒性作用,可显著诱导细胞凋亡(p<0.0001)。此外,与未处理的细胞相比,GAA下调了Nalm-6细胞中miR-17-5p和miR-181b的表达(P=0.0067和P=0.0014)。结论:基于目前的研究结果,GAA作为一种治疗血液系统恶性肿瘤的天然试剂值得进一步研究。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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