Effect of Yangzheng Sanjie Decoction on Proliferation， Apoptosis and ERK Pathway of Human Gastric Cancer MKN-45 Cells

Zhen-zhen XIAO; Hai-xia DENG; Geng-xin CHEN; Chao-fen QIU; Ao-qiang ZHOU

doi:10.13422/j.cnki.syfjx.20190327

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Effect of Yangzheng Sanjie Decoction on Proliferation， Apoptosis and ERK Pathway of Human Gastric Cancer MKN-45 Cells

Pharmacology | 更新时间：2021-02-09

- Effect of Yangzheng Sanjie Decoction on Proliferation， Apoptosis and ERK Pathway of Human Gastric Cancer MKN-45 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 3, Pages: 39-44(2019)
- 作者机构：
  
  广州中医药大学　第二附属医院，广州　 510120
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190327
  CLC： R22;R242;R2-031;R273;R285.5
- Received：29 August 2018，
  
  Published Online：16 November 2018，
  
  Published：05 February 2019
- 稿件说明：
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Zhen-zhen XIAO, Hai-xia DENG, Geng-xin CHEN, et al. Effect of Yangzheng Sanjie Decoction on Proliferation， Apoptosis and ERK Pathway of Human Gastric Cancer MKN-45 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(3): 39-44.
DOI：

Zhen-zhen XIAO, Hai-xia DENG, Geng-xin CHEN, et al. Effect of Yangzheng Sanjie Decoction on Proliferation， Apoptosis and ERK Pathway of Human Gastric Cancer MKN-45 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(3): 39-44. DOI： 10.13422/j.cnki.syfjx.20190327.

摘要

目的:

探讨养正散结汤对人胃癌MKN-45细胞增殖、凋亡及细胞外信号调节激酶(extracellular signal-regulated kinase

ERK)通路的影响。

方法:

将养正散结汤组(0.5，1，1.5，2，2.5，3，3.5 g·L

-1

)作用于胃癌MKN-45细胞，设置空白组，分别孵育24，48，72 h后采用细胞增殖活性检测方法(CCK-8)检测细胞增殖情况；用养正散结汤组(0.4，0.8 g·L

-1

)处理细胞，运用平板克隆形成实验观察细胞的克隆形成能力；用4，8 g·L

-1

养正散结汤干预细胞，利用流式细胞仪检测细胞的凋亡情况；用2，4，8 g·L

-1

养正散结汤处理细胞，采用蛋白免疫印迹法(Western blot)检测ERK表达及其磷酸化水平。

结果:

与空白组比较，养正散结汤组能明显抑制MKN-45细胞的增殖，处理24，48 h后，养正散结汤从2 g·L

-1

开始，处理72 h后，从1.5 g·L

-1

开始，细胞存活率逐渐降低，呈明显浓度依赖性(

0.05，

0.01)。经不同质量浓度的养正散结汤干预48 h后，养正散结汤组细胞克隆形成率显著低于空白组(

0.01)，呈一定的量效关系，0.8 g·L

-1

养正散结汤干预后，MKN-45细胞集落基本无法形成；养正散结汤组细胞凋亡率明显高于空白组(

0.05，

0.01)；干预12 h后，与空白组比较，养正散结汤从4 g·L

-1

开始下调MKN-45细胞中ERK蛋白的磷酸化水平(

0.01)。

结论:

养正散结汤能抑制人胃癌细胞MKN-45的增殖，促进其凋亡，其作用机制可能与抑制ERK的磷酸化有关。

Abstract

Objective:

To investigate the effect of Yangzheng Sanjie decoction on proliferation

apoptosis and extracellular signal-regulated kinase (ERK) pathway of human gastric cancer MKN-45 cells.

Method:

Gastric cancer cell line MKN-45 was treated for 24

72 h with Yangzheng Sanjie decoction (0.5

1.5

2.5

3.5 g·L

-1

); cell proliferation was measured by cell counting kit-8 (CCK-8); cell colony forming ability was observed by the plate cloning experiment after intervention with Yangzheng Sanjie decoction (0.4

0.8 g·L

-1

); MKN-45 cells was treated with 4

8 g·L

-1

and then cell apoptosis was detected by flow cytometry; the expression of ERK and its phosphorylation level were detected by Western blot assay after treatment with 2

8 g·L

-1

Result:

Compared with the blank group

Yangzheng Sanjie decoction could significantly inhibit the proliferation of MKN-45 cells. After treatment for 24

48 h

Yangzheng Sanjie decoction started from 2 g·L

-1

and after treatment for 72 h

it started from 1.5 g·L

-1

the cell viability gradually decreased in a concentration-dependent manner (

0.05

0.01). After 48 hours of intervention with different concentrations of Yangzheng Sanjie decoction

the cell clone formation rate of Yangzheng Sanjie decoction was significantly lower than that of the blank group with a dose-response relationship (

0.01). After the intervention with Yangzheng Sanjie decoction at a concentration of 0.8 g·L

-1

cell colonies could not be formed; the apoptosis rate of Yangzheng Sanjie decoction was significantly higher than that of the blank group (

0.05

0.01); after 12 h of intervention

compared with the blank group

Yangzheng Sanjie decoction started from 4 g·L

-1

and the phosphorylation level of ERK protein in MKN-45 cells was down-regulated (

0.01).

Conclusion:

Yangzheng Sanjie decoction can inhibit the proliferation of human gastric cancer cell line MKN-45 and promote its apoptosis. The mechanism may be related to the inhibition of phosphorylation of ERK.

关键词

Keywords

references

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