Effect of Yanghetang on Apoptosis of Triple Negative Breast Cancer Cell Line MDA-MB-231 Based on Egr1/p21 Signaling Pathway

Kang-le LI; Xiao-qian YANG; Yu ZHANG; Shuo YANG; Zhong-bo ZHU; Jian-wei DOU

doi:10.13422/j.cnki.syfjx.20200424

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Effect of Yanghetang on Apoptosis of Triple Negative Breast Cancer Cell Line MDA-MB-231 Based on Egr1/p21 Signaling Pathway

Pharmacology | 更新时间：2021-02-09

- Effect of Yanghetang on Apoptosis of Triple Negative Breast Cancer Cell Line MDA-MB-231 Based on Egr1/p21 Signaling Pathway
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 8, Pages: 62-67(2020)
- 作者机构：
  
  1.西安交通大学　药学院，西安　 710061
  2.西安交通大学　第二附属医院，西安　 710004
  3.甘肃中医药大学　基础医学院，兰州　 730000
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200424
  CLC： R22; R242; R2-031;R285.5
- Received：08 July 2019，
  
  Published Online：04 November 2019，
  
  Published：20 April 2020
- 稿件说明：
移动端阅览
Kang-le LI, Xiao-qian YANG, Yu ZHANG, et al. Effect of Yanghetang on Apoptosis of Triple Negative Breast Cancer Cell Line MDA-MB-231 Based on Egr1/p21 Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(8): 62-67.
DOI：

Kang-le LI, Xiao-qian YANG, Yu ZHANG, et al. Effect of Yanghetang on Apoptosis of Triple Negative Breast Cancer Cell Line MDA-MB-231 Based on Egr1/p21 Signaling Pathway[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(8): 62-67. DOI： 10.13422/j.cnki.syfjx.20200424.

摘要

目的：

以早期生长反应基因1(Egr1)/细胞周期蛋白依赖性抑制剂(p21)信号通路为基础，探讨古方阳和汤含药血清对人三阴性乳腺癌细胞MDA-MB-231凋亡的影响。

方法：

制备阳和汤大鼠含药血清，体外培养三阴性乳腺癌MDA-MB-231细胞。设空白组，阳和汤高、中、低剂量组(23.4，11.7，5.85 g·kg

-1

)，分别干预MDA-MB-231细胞24，48，72 h，细胞增殖检测(CCK-8)法检测各组细胞增殖情况。设空白组，阳和汤高、中、低剂量组(23.4，11.7，5.85 g·kg

-1

)，干预MDA-MB-231细胞48 h，采用流式细胞仪检测各组细胞凋亡情况以及细胞周期分布情况；实时荧光定量聚合酶链式反应(Real-time PCR)检测各组细胞Egr1，p21 mRNA的表达情况；蛋白免疫印迹法(Western blot)检测各组细胞Egr1，p21蛋白的表达情况。

结果：

阳和汤干预MDA-MB-231细胞24，48，72 h，与空白组相比，阳和汤高、中剂量组能显著抑制MDA-MB-231细胞增殖(

0.01)。阳和汤干预MDA-MB-231细胞48 h，与空白组相比，阳和汤高、中剂量组细胞的凋亡率明显增加(

0.05，

0.01)；阳和汤高、中剂量组细胞周期G

期所占比例降低，G

/M期所占比例升高(

0.05，

0.01)；阳和汤高、中剂量组细胞Egr1，p21 mRNA和蛋白表达均增加(

0.05，

0.01)。

结论：

阳和汤可激活MDA-MB-231细胞中Egr1/p21信号通路，增加Egr1，p21的表达，引起G

/M细胞周期阻滞，从而诱导MDA-MB-231细胞凋亡。

Abstract

Objective:

To investigate the effect of ancient recipe Yanghetang and its drug-contained serum on apoptosis of triple negative breast cancer cell line MDA-MB-231 based on the signal pathway of early growth response gene 1 (Egr1)/cyclin dependent inhibitor (p21).

Method:

The drug-containing serum of Yanghetang was prepared from rats

and MDA-MB-231 cells were cultured

in vitro

. The blank control group was set

and MDA-MB-231 cells in Yanghetang high

middle

and low dose groups (23.4

11.7

5.85 g·kg

-1

) were intervened for 24

72 h

respectively. After that

the cell counting kit-8(CCK-8) method was used to detect the cell proliferation of each group. The blank control group was set

while MDA-MB-231 cells in Yanghetang high

middle

and low dose groups (23.4

11.7

5.85 g·kg

-1

) were treated for 48 h

and then flow cytometry was used to detect the apoptosis of each group and the distribution of cell cycle. The expression of Egr1 and p21 mRNA in each group was detected by quantificational Real-time polymerase chain reaction (Real-time PCR)

while the expression of Egr1 and p21 protein in each group was detected by Western blot.

Result:

After MDA-MB-231 cells were intervened by Yanghetang for 24

72 h

MDA-MB-231 cell proliferation was significantly inhibited in Yanghetang high and middle dose groups as compared with the blank control group (

0.01). After MDA-MB-231 cells were intervened by Yanghetang for 48 h

the apoptosis was significantly increased in Yanghetang high and middle dose groups as compared with the blank control group (

0.05

0.01). In the Yanghetang high

middle dose groups

the proportion of cell cycle G

phase decreased

and the proportion of G

/M phase increased (

0.05

0.01). The mRNA and protein expressions of Egr1

p21 were increased in Yanghetang high and middle groups (

0.05

0.01).

Conclusion:

Yanghetang can activate Egr1/p21 signaling pathway in MDA-MB-231 cells

increase the expression of Egr1 and p21

and cause G

/M cell cycle arrest

thereby inducing apoptosis of MDA-MB-231 cells.

关键词

Keywords

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