Effect and Mechanism of Curdione on Migration and  Invasion of Breast Cancer HCC1937 Cells

Xue-ran SUN; Ke YANG; Ling-ling LYU; Jing-xian CHEN; Yuan-biao GUO; Ming RUAN; Hai-feng YING; Xiao-heng SHEN; Lan ZHENG

doi:10.13422/j.cnki.syfjx.20190326

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Effect and Mechanism of Curdione on Migration and Invasion of Breast Cancer HCC1937 Cells

Pharmacology | 更新时间：2021-02-09

- Effect and Mechanism of Curdione on Migration and Invasion of Breast Cancer HCC1937 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 25, Issue 3, Pages: 66-73(2019)
- 作者机构：
  
  上海交通大学　医学院　附属瑞金医院，上海　 200025
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20190326
  CLC： R22;R242;R2-031;R273
- Received：03 September 2018，
  
  Published Online：16 November 2018，
  
  Published：05 February 2019
- 稿件说明：
移动端阅览
Xue-ran SUN, Ke YANG, Ling-ling LYU, et al. Effect and Mechanism of Curdione on Migration and Invasion of Breast Cancer HCC1937 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(3): 66-73.
DOI：

Xue-ran SUN, Ke YANG, Ling-ling LYU, et al. Effect and Mechanism of Curdione on Migration and Invasion of Breast Cancer HCC1937 Cells[J]. Chinese journal of experimental traditional medical formulae, 2019, 25(3): 66-73. DOI： 10.13422/j.cnki.syfjx.20190326.

摘要

目的:

探讨莪术二酮对人乳腺癌HCC1937细胞的迁移和侵袭能力的影响及其作用机制。

方法:

体外培养HCC1937细胞，分别以12.5，25，50，100，200，400 μmol·L

-1

莪术二酮处理24，48 h，设空白组，用细胞计数试剂盒法(CCK-8)检测莪术二酮对HCC1937细胞活力的影响；设立空白组，莪术二酮12.5，25，50 μmol·L

-1

组，细胞黏附实验检测莪术二酮对HCC1937细胞黏附力的影响；划痕愈合实验检测莪术二酮对HCC1937细胞迁移力的影响；transwell小室体外迁移和侵袭实验检测莪术二酮对HCC1937细胞迁移和侵袭力的影响；蛋白免疫印迹法(Western blot)检测莪术二酮对HCC1937细胞中丝裂原活化蛋白激酶(MAPK)和蛋白激酶B(Akt)信号通路的调节作用以及对基质金属蛋白酶-2(MMP-2)，基质金属蛋白酶-9(MMP-9)蛋白表达水平的影响；实时荧光定量PCR(Real-time PCR)检测莪术二酮对HCC1937细胞中MMP-2，MMP-9 mRNA表达水平的影响。

结果:

与空白组比较，莪术二酮12.5，25，50 μmol·L

-1

组对HCC1937细胞活力无明显影响，莪术二酮100，200，400 μmol·L

-1

组对HCC1937细胞活力有明显的抑制作用(

0.05，

0.01)，且呈时间-浓度依赖性；与空白组比较，莪术二酮12.5，25，50 μmol·L

-1

组能明显降低HCC1937细胞的黏附率、迁移率、迁移和侵袭细胞数(

0.05，

0.01)；与空白组比较，莪术二酮12.5，25，50 μmol·L

-1

组能明显下调MAPK和Akt信号通路上关键蛋白蛋白激酶(ERK)，c-Jun氨基末端激酶(JNK)和Akt的磷酸化水平(

0.01)，MMP-2，MMP-9蛋白和mRNA表达显著下降(

0.01)。

结论:

莪术二酮能明显抑制人乳腺癌HCC1937细胞的迁移和侵袭能力，其机制可能与通过下调MAPK和Akt信号通路上关键蛋白ERK

JNK

Akt的磷酸化水平，进而使MMP2，MMP-9表达量下降有关。

Abstract

Objective:

To investigate effect of curdione on the migration and invasion of human breast cancer HCC1937 cells and its mechanism.

Method:

HCC1937 cells were cultured

in vitro

and treated with curdione at various doses (0

12.5

100

200

400 μmol·L

-1

) for 24

48 h

the cell viability was detected by cell counting kit-8 method. curdione groups (12.5

50 μmol·L

-1

) and blank group were established. The effect of curdione on the adhesion of HCC1937 cells was detected by the cell adhesion assay. The effect of curdione on migration of HCC1937 cells was detected by wound healing assay. The effect of curdione on the migration and invasion of HCC1937 cells were detected by transwell chamber assay. The effect of curdione on regulation of mitogen-activated protein kinase(MAPK)and protein kinase B(Akt)signaling pathways and the protein expressions of matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) of HCC1937 cells were detected by the Western blot analysis. Effect of curdione on mRNA expressions of MMP-2 and MMP-9 of HCC1937 cells were detected by Real-time PCR.

Result:

Compared with the blank group

curdione (12.5

50 μmol·L

-1

) groups had no significant effect on cell viability

but a remarkable effect on cell viability HCC1937 cells

and cell viability was gradually decreased with the increase of the concentration of curdione (

0.05

0.01) in a time and dose-dependent manner. Compared with blank group

curdione groups (12.5

50 μmol·L

-1

) had a significant effect on cell adhesion rate

migration rate and invasion rate of HCC1937 cells (

0.05

0.01). Compared with the blank group

curdione groups (12.5

50 μmol·L

-1

) could down-regulate phosphorylation levels of key proteins extracellular regulated protein kinases(ERK)

c-Jun

-terminal kinase(JNK)

Akt on MAPK and Akt signaling pathways (

0.01)

as well as the protein and mRNA expressions of MMP-2 and MMP-9 of HCC1937 cells.

Conclusion:

curdione can inhibit the migration and invasion of human breast cancer HCC1937 cells

and the mechanism may be related to down-regulation of phosphorylation levels of key proteins ERK

JNK

Akt on MAPK and Akt signaling pathways

so as to reduce the expressions of MMP2 and MMP-9.

关键词

Keywords

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