Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/<italic style="font-style: italic">β</italic>-Catenin Pathway

Yi-yun HE; Yi-nan YIN; Hai-rong ZENG; Li-li QI

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Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/β-Catenin Pathway

Pharmacology | 更新时间：2021-05-17

- Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/β-Catenin Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 27, Issue 11, Pages: 83-88(2021)
- 作者机构：
  
  1.上海中医药大学附属岳阳中西医结合医院，上海 200437
  2.上海市普陀区中心医院，上海 200062
- 作者简介：
- 基金信息：
- DOI：
  CLC： R2-0;R22;R285.5;R289
- Received：06 February 2021，
  
  Published Online：09 April 2021，
  
  Published：05 June 2021
- 稿件说明：
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何艺韵,殷亦男,曾海荣等.柴胡皂苷A通过Wnt/β-catenin通路改变A549/DDP细胞耐药性[J].中国实验方剂学杂志,2021,27(11):83-88.

HE Yi-yun,YIN Yi-nan,ZENG Hai-rong,et al.Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/β-Catenin Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(11):83-88.
何艺韵,殷亦男,曾海荣等.柴胡皂苷A通过Wnt/β-catenin通路改变A549/DDP细胞耐药性[J].中国实验方剂学杂志,2021,27(11):83-88. DOI：

HE Yi-yun,YIN Yi-nan,ZENG Hai-rong,et al.Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/β-Catenin Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(11):83-88. DOI：

摘要

目的

探讨柴胡皂苷A（SSA）逆转人肺癌顺铂（DDP）耐药株A549/DDP的作用及机制。

方法

细胞增殖与活性检测（CCK-8）法检测A549和A549/DDP对DDP的耐药性及SSA对A549和A549/DDP细胞活力的影响；流式细胞术检测A549/DDP凋亡率和细胞中活性氧（ROS）的变化；实时荧光定量聚合酶链反应（Real-time PCR）检测C-myc，B淋巴细胞瘤-2（Bcl-2）和含半胱氨酸的天冬氨酸蛋白水解酶-3（Caspase-3） mRNA的表达情况；TopFish双荧光素酶报告基因检测

-连环蛋白（

-catenin）的转录活性；蛋白免疫印迹法（Western blot）法检测A549/DDP细胞中

-catenin蛋白的变化。

结果

CCK-8检测结果显示，A549/DDP的耐药性是A549的12.82倍，差异具有统计学意义（

0.05），SSA能够抑制A549的细胞活力，其半数抑制浓度（IC

）为34.9 μmol·L

-1

。SSA抑制 A549/DDP细胞的活力并具有浓度依赖性，在20 μmol·L

-1

浓度时，对A549/DDP的抑制率

10%，选择20 μmol·L

-1

作为逆转浓度；流式结果显示，与空白组比较，DDP组A549/DDP的凋亡比例明显升高，细胞内活性氧的累积明显增加（

0.05），与DDP组比较，SSA+DDP组A549/DDP的凋亡比例明显升高，细胞内活性氧的累积明显增加；与空白组比较，DDP组A549/DDP细胞中C-myc，Bcl-2 mRNA表达明显降低，Caspase-3 mRNA表达明显升高（

0.05），与DDP组比较，SSA+DDP组C-myc，Bcl-2 mRNA表达明显降低，Caspase-3 mRNA表达明显升高（

0.05）；与空白组比较，DDP组

-catenin的转录活性明显降低（

0.05），与DDP组比较，SSA+DDP组

-catenin的转录活性明显降低（

0.05）；与空白组和DDP组比较，SSA+DDP组

-catenin蛋白表达明显降低，差异有统计学意义（

0.05）。

结论

柴胡皂苷A可以提高A549/DDP细胞对DDP的敏感性，其作用机制可能与抑制Wnt/

-catenin信号通路激活有关。

Abstract

Objective

To investigate the effect and mechanism of saikosaponin A （SSA） on the reversal of cisplatin （DDP） resistance in human lung cancer cell line A549/DDP.

Methods

The resistance of A549 and A549/DDP cells to DDP and the inhibitory effects of SSA against the proliferation of A549 and A549/DDP cells were detected using cell counting kit-8 （CCK-8）. The apoptosis rates of A549/DDP cells treated with SSA or DDP or SSA combined with DDP and the changes in reactive oxygen species （ROS） were determined by flow cytometry. The mRNA expression levels of C-myc， B-cell lymphoma 2 （Bcl-2） and cysteinyl aspartate-specific protease-3 （Caspase-3） were detected by real-time polymerase chain reaction （Real-time PCR）， followed by the determination of

-catenin transcriptional activity using the TopFish dual-luciferase reporter assay system and the measurement of

-catenin protein expression in A549/DDP cells by Western blot.

Results

The results of CCK-8 assay showed that the DDP resistance of A549/DDP cells was 12.82 times that of A549 cells （

0.05）. SSA inhibited the viability of A549 cells with the half maximal inhibitory concentration （IC

） being 34.9 μmol·L

-1

， and also suppressed the viability of A549/DDP cells in a concentration-dependent manner. Since the inhibition rate of 20 μmol/L SSA against A549/DDP cells was less than 10%， the reversal concentration was set at 20 μmol/L. Flow cytometry revealed that compared with the control， DDP alone increased the apoptosis rate of A549/DDP cells （

0.05）， stimulated the accumulation of intracellular ROS （

0.05）， down-regulated the mRNA expression levels of C-myc and Bcl-2 in A549/DDP cells， up-regulated Caspase-3 mRNA expression， and reduced the transcriptional activity of

-catenin （

0.05）. Compared with the DDP group， the SSA+DDP group exhibited obviously increased apoptosis of A549/DDP cells， enhanced accumulation of intracellular ROS， down-regulated C-myc and Bcl-2 mRNA expression， up-regulated Caspase-3 mRNA expression （

0.05）， and weakened

-catenin transcription （

0.05）. DDP combined with SSA better decreased the

-catenin protein expression in contrast to that of control or DDP （

0.05）.

Conclusions

SSA enhances the sensitivity of A549/DDP cells to DDP possibly by inhibiting the activation of Wnt/

-catenin pathway.

关键词

Keywords

references

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Saikosaponin A Changes Drug Resistance of A549/DDP Cells Through Wnt/β-Catenin Pathway

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摘要

Abstract

关键词

Keywords

references