熊果酸通过调节 miR-373-3p 对肺癌 A549 细胞凋亡和迁移的影响
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河南中医药大学,郑州 450046

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R-33


Effects of ursolic acid on apoptosis and migration of A549 lung cancer cells by regulating miR-373-3p
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Affiliation:

Henan University of Chinese Medicine, Zhengzhou 450046, China

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    摘要:

    目的 探讨熊果酸通过调节 miR-373-3p 对肺癌 A549 细胞凋亡和迁移的影响。 方法 MTT 法检测熊果酸对肺癌 A549 细胞的毒性作用;AnnexinⅤ- FITC/ PI 双染法、流式细胞术检测熊果酸作用于转染 miR-373-3p 的 A549 细胞后对 A549 细胞凋亡影响;实时荧光定量 PCR 法分别检测熊果酸加药组和转染 miR-373-3p mimics 组中 A549 细胞 miR-373-3p 表达程度;RT-PCR 和免疫印迹法检测熊果酸作用下 miR-373-3p 调控的下游靶基因 TXNIP mRNA 及蛋白的表达;细胞划痕愈合实验检测 miR-373-3p 细胞迁移能力。 结果 熊果酸显著降低肺癌 A549 细胞的生长活力(P<0. 05),IC 50 为 60 μmol / L;瞬时转染 miR-373-3p mimics 可以上调 A549 细胞中 miR-373- 3p 的表达(24 h,11. 367±2. 120;48 h,12. 167±1. 108);熊果酸作用于转染 miR-373-3p mimics 的 A549 细胞后,能够显著提高细胞中 miR-373-3p mimics 的表达水平( 24 h,16. 787 ± 3. 109;48 h,16. 980 ± 3. 106);并且 miR-373-3p mimics 下游预测的目标基因 TXNIP mRNA(10. 757±0. 79)及蛋白表达量(0. 8210±0. 043)均明显上升;FCM 结果显示:miR-373-3p 能够促进肺癌 A549 细胞凋亡(46. 20±5. 970)。 细胞转染 miR-373-3p mimics 后,迁移能力明显受到抑制(P<0. 001)。 结论 熊果酸能够通过促进肺癌 A549 细胞凋亡抑制其迁移,其作用机制可能是通过上调 miR- 373-3p 表达来完成的。

    Abstract:

    Objective To investigate the effect of ursolic acid on apoptosis and migration of lung cancer A549 cells by regulating miR-373-3p. Methods We used an MTT assay to detect the toxic effects of ursolic acid on A549 lung cancer cells. AnnexinⅤ-FITC/ PI double staining and flow cytometry were used to detect the effect of ursolic acid on A549 cells transfected with miR-373-3p. Real-time fluorescence quantitative PCR was used to detect the expression level of miR- 373-3p in A549 cells in the ursolic acid dosing group and the transfected miR-373-3p mimics group. RT-PCR and Western blot were used to detect mRNA and the protein expression of the downstream target gene, TXNIP, which is regulated by miR-373-3p under the action of ursolic acid. The scratch text was used to evaluate miR-373-3p cell migration ability. Results Ursolic acid significantly decreased the growth viability of A549 lung cancer cells (P<0. 05), with an IC50 of 60 μmol / L. Transiently transfected miR-373-3p mimics up-regulated the expression of miR-373-3p in A549 cells ( 24 h, 11. 367± 2. 120; 48 h, 12. 167 ± 1. 108), and ursolic acid significantly increased the expression level of miR-373-3p mimics in A549 cells transfected with miR-373-3p mimics (24 h, 16. 787±3. 109; 48 h, 16. 980±3. 106). mRNA for the target gene, TXNIP, (10. 757± 0. 79) and protein expression ( 0. 8210± 0. 043) predicted by miR-373-3p mimics were significantly increased. FCM result showed that miR-373-3p promoted apoptosis of A549 lung cancer cells ( 46. 20 ± 5. 970), and after being transfected with miR-373-3p mimics, the cells’ migration ability was significantly inhibited (P< 0. 001). Conclusions Ursolic acid can promote the apoptosis of A549 lung cancer cells and inhibit their migration. The mechanism may involve up-regulation of miR-373-3p expression.

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陈换换,张小莉,桑晓林,冯 龙,于莹莹,宋超杰,李志慧.熊果酸通过调节 miR-373-3p 对肺癌 A549 细胞凋亡和迁移的影响[J].中国比较医学杂志,2021,31(3):36~42.

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  • 收稿日期:2020-06-07
  • 在线发布日期: 2021-04-30
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