Role of G protein-coupled estrogen receptor in tamoxifen-resistant breast cancer MCF-7 cells

朱庆; 伍俊仪; 袁杰; 杨海兵; 陈茂山; 杨光伦

您当前的位置：

首页 >

文章列表页 >

Role of G protein-coupled estrogen receptor in tamoxifen-resistant breast cancer MCF-7 cells

更新时间：2026-01-09

- Role of G protein-coupled estrogen receptor in tamoxifen-resistant breast cancer MCF-7 cells
- Journal of Army Medical University Vol. 37, Issue 22, Pages: 2249-2254(2015)
- 作者机构：
- 作者简介：
- 基金信息：
- DOI：
  CLC： R73-36;R730.23;R737.9
- Published：2015
- 稿件说明：
移动端阅览
朱庆,伍俊仪,袁杰,杨海兵,陈茂山,杨光伦.新型雌激素受体GPER在乳腺癌MCF-7细胞他莫昔芬耐药中的作用及机制[J].陆军军医大学学报(原第三军医大学学报),2015,37(22):2249-2254.

　Zhu Qing,Wu Junyi,Yuan Jie,Yang Haibing,Chen Maoshan,Yang Guanglun.Role of G protein-coupled estrogen receptor in tamoxifen-resistant breast cancer MCF-7 cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(22):2249-2254.
朱庆,伍俊仪,袁杰,杨海兵,陈茂山,杨光伦.新型雌激素受体GPER在乳腺癌MCF-7细胞他莫昔芬耐药中的作用及机制[J].陆军军医大学学报(原第三军医大学学报),2015,37(22):2249-2254. DOI：

　Zhu Qing,Wu Junyi,Yuan Jie,Yang Haibing,Chen Maoshan,Yang Guanglun.Role of G protein-coupled estrogen receptor in tamoxifen-resistant breast cancer MCF-7 cells[J].J Amry Med Univ (J Third Mil Med Univ),2015,37(22):2249-2254. DOI：

摘要

目的研究乳腺癌他莫昔芬耐药细胞MCF-7R中新型雌激素受体（G protein-coupled estrogen receptor， GPER）对促凋亡蛋白Bim的影响及其机制。方法倒置显微镜下观察MCF-7和MCF-7R经过浓度为1 μmol/L的他莫昔芬（tamoxifen

TAM）处理后细胞形态的变化；qRT-PCR检测人乳腺癌细胞MCF-7及其他莫昔芬耐药株MCF-7R中促凋亡蛋白Bim（Bcl-2L11）的mRNA表达水平，Western blot检测两种细胞株Bim蛋白表达水平和两种细胞中总的GPER和膜上GPER的表达水平；两种细胞中分别以（无水乙醇、TAM、GPER特异性抑制剂G15+TAM、GPER特异性激动剂G1）处理后，检测Bim的蛋白表达水平；MCF-7R细胞中分别抑制PI3K-Akt及MAPK/Erk两条信号通路后，再加入TAM，检测Bim的蛋白水平；流式细胞术检测细胞凋亡。结果倒置显微镜下发现MCF-7细胞经TAM处理后细胞大部分变透亮且悬浮，而MCF-7R无明显变化；MCF-7R细胞中促凋亡蛋白Bim的mRNA水平及蛋白水平较MCF-7都降低(P0.05)；耐药细胞中总GPER的蛋白表达量较MCF-7无明显变化而膜上的表达量升高(P0.05)；MCF-7经TAM处理后，Bim蛋白表达量增加（P0.05），而MCF-7R中却无明显变化，但在MCF-7R中加入GPER特异性抑制剂G15后再用TAM处理，Bim蛋白水平增加（P0.05）；耐药细胞中PI3K/AKT及MAPK/Erk信号通路处于活化状态，抑制MAPK/Erk信号通路后再加入TAM，耐药细胞中促凋亡蛋白Bim的表达量增加（P0.05）；在MCF-7中加入TAM后细胞凋亡率明显升高（P0.05），MCF-7R经TAM处理后无明显差异，但抑制GPER或Erk信号通路后，再加入TAM处理，细胞凋亡率明显增高（P0.05）。 结论乳腺癌TAM耐药细胞MCF-7R中TAM作为GPER的激动剂，活化信号通路MAPK/Erk抑制Bim表达。抑制GPER或MAPK/Erk信号通路，可使耐药细胞对TAM恢复敏感性。

Abstract

Objective To investigate the effect of G protein-coupled estrogen receptor (GPER) on pro-apoptotic protein Bim in tamoxifen (TAM)-resistant breast cancer cells MCF-7R and related mechanism. Methods Human breast cancer cells MCF-7 and the MCF-7R cells were treated with 1 μmol/L TAM

and the cell morphological change was observed with an inverted microscope. The Bim mRNA expression was detected by quantitative real-time PCR (qRT-PCR) in the MCF-7 cells and MCF-7R cells. The expression of Bim protein

total GPER

and GPER in the cell membrane was detected by Western blotting. After the two kinds of cells were treated with ethanol

TAM

TAM+G15 (GPER specific inhibitor)

and G1 (GPER specific agonist)

the Bim protein expression was analyzed by Western blotting. After inhibiting the PI3K/AKT and MAPK/Erk signaling pathways in the MCF-7R cells

TAM was added

and then the Bim protein expression was assayed. Flow cytometry was adopted to detect cell apoptosis. Results After treatment with TAM

most of MCF-7 cells became bright and were suspended

while the MCF-7R cells had no obvious changes. The levels of Bim mRNA and protein in the MCF-7R cells were obviously lower than those in the MCF-7 cells (P0.05). The total GPER had no significant difference between the 2 kinds of cells

but GPER in the cell membrane was increased in the MCF-7R cells (P0.05). The expression of Bim protein was increased in the MCF-7 cells treated with TAM (P0.05)

but not in the MCF-7R cells. However

Bim protein was increased in the MCF-7R cells treated with G15 and then TAM (P0.05). The PI3K/AKT and MAPK/Erk signaling pathways stayed in the activated stage in the MCF-7R cells. When TAM was added after inhibiting MAPK/Erk pathway rather than PI3K/AKT pathway

the Bim protein was significantly increased (P0.05).TAM could promote the apoptosis of the MCF-7 cells

but not the MCF-7R cells. However

the apoptosis of MCF-7R cells was increased after GPER or MAPK/Erk signaling pathway was inhibited and then TAM was added (P0.05). Conclusion TAM as GPER agonist can activate the MAPK/Erk signaling pathway and inhibit the Bim expression in the MCF-7R cells. The MCF-7R cells are sensitive to TAM again through inhibiting GPER or MAPK/Erk signaling pathway.

关键词

Keywords

references

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

High expression of kinesin KIF18B in estrogen receptor-positive breast cancer enhances cell proliferation and apoptosis resistance and predicts poor prognosis

Identification of epithelial-mesenchymal transition phenotype in tamoxifen-resistant MCF-7 cells

A novel estrogen receptor expression stratification (low/moderate/high) based on Chinese population provides more accurate prognosis prediction for HR⁺/HER2^- early breast cancer patients

Neuroprotective effects and mechanisms of osteocalcin in an Alzheimer’s disease cell model

Mechanism of abnormal specification of spermatogonial stem cells after Rb1 knockout in mitotic-arrested prospermatogonia

Related Author

黄飞翔

向廷秀

任国胜

袁杰

杨丽

陈茂山

朱庆

付伟杰

Related Institution

Department of Breast and Thyroid Surgery， First Affiliated Hospital， Army Medical University （Third Military Medical University）

Graduate School of Guangxi University of Chinese Medicine

Department of Neurology

Center for Integrated Traditional Chinese and Western Medicine， Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine

Department of Encephalopathy， Pingnan County Hospital of Chinese Medicine

⁰