Role of yes-associated protein in resistance of HER2<sup>+</sup> breast cancer to lapatinib

Peng CHEN; Ting XU; Dan GUO; Jiayi HUANG

doi:10.16016/j.1000-5404.201911189

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Role of yes-associated protein in resistance of HER2⁺ breast cancer to lapatinib

Basic Medicine | 更新时间：2026-01-09

- Role of yes-associated protein in resistance of HER2⁺ breast cancer to lapatinib
- Journal of Third Military Medical University Vol. 42, Issue 7, Pages: 692-698(2020)
- 作者机构：
  
  1.400016 重庆，重庆医科大学基础医学院病理生理学教研室
  2.400016 重庆，重庆医科大学附属第二医院三腺外科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.201911189
  CLC： R737.9;R915;R979.1
- Received：22 November 2019，
  
  Revised：2019-12-17，
  
  Published：15 April 2020
- 稿件说明：
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Peng CHEN, Ting XU, Dan GUO, et al. Role of yes-associated protein in resistance of HER2⁺ breast cancer to lapatinib[J]. Journal of Third Military Medical University, 2020, 42(7): 692-698.
DOI：

Peng CHEN, Ting XU, Dan GUO, et al. Role of yes-associated protein in resistance of HER2⁺ breast cancer to lapatinib[J]. Journal of Third Military Medical University, 2020, 42(7): 692-698. DOI： 10.16016/j.1000-5404.201911189.

摘要

目的

建立乳腺癌拉帕替尼耐药细胞株，探讨Hippo通路中YAP蛋白在HER2阳性乳腺癌细胞拉帕替尼耐药中的作用及机制。

方法

采用逐步增加剂量与高浓度反复间歇诱导乳腺癌细胞株Skbr3对拉帕替尼耐药，耐药株命名为Skbr3-LR。用拉帕替尼同时处理亲本细胞和耐药细胞株，CCK-8法检测其耐药指数；结晶紫染色和平板克隆形成实验检测细胞的存活状况；定量PCR检测上皮-间质化改变。定量PCR及Western blot检测YAP、TAZ的表达。以维替泊芬阻断耐药细胞株Skbr3-LR中YAP下游通路的激活，CCK-8法检测其细胞活性的改变并计算IC

值的变化；结晶紫染色检测细胞存活数量的变化。定量PCR及Western blot检测YAP表达的改变。

结果

与亲本细胞比较，结晶紫染色和平板克隆形成实验结果均显示，耐药株存活细胞数量明显增多。亲本细胞耐药指数为3.61 μmol/L，耐药株耐药指数上升约8倍，为29.26 μmol/L。PCR结果显示，与亲本细胞比较，耐药细胞株的上皮标志物E-cadherin表达降低，而间质标志物Twist、Vinemtin和N-cadherin表达上升，YAP/TAZ表达上升，差异均有统计学意义(

0.05)。Western blot结果显示，耐药细胞株中总YAP和磷酸化YAP表达均较亲本细胞显著上升(

0.05)。以维替泊芬阻断耐药细胞株YAP下游通路的激活后，其耐药指数为2.98 μmol/L，较未处理的Skbr3-LR耐药指数下降约4倍；其存活细胞数量明显降低，YAP表达也明显降低(

0.05)。

结论

成功建立了拉帕替尼获得性耐药的乳腺癌细胞模型，其耐药机制可能与Hippo信号通路下游的关键效应因子YAP表达异常有关。

Abstract

Objective

To construct a lapatinib-resistant breast cancer cell line

and to study the role and underlying mechanism of yes-associated protein (YAP) in Hippo signaling pathway in the process.

Methods

The drug-resistant cell model of human breast cancer was established by gradually increasing the dose of drug and repeated intermittent high concentration inducing. After lapatinib treatment

the parent cell line Skbr3 and the drug-resistant cell line Skbr3- LR were detected by CCK-8 assay for drug resistance index

crystal violet staining and plate colony for cell survival

and quantitative PCR for the expression of epithelial mesenchymal transformation (EMT) markers. The expression of YAP/TAZ at mRNA and protein levels were measured by quantitative PCR and Western blotting

respectively. After verteporfin was used to block the downstream pathway of YAP in Skbr3-LR cells

CCK-8 assay was employed to measure the cell viability to obtain IC

cell crystal violet staining was performed to count the number of survival cells

and the change of YAP expression was be detected by quantitative PCR and Western blotting.

Results

The results of crystal violet staining and plate colony showed that there were more survival cells in the Skbr3-LR cells when compared with the parent Skbr3 cells. The drug resistance index was 3.61 μmol/L in the parent Skbr3 cells

and was increased by 8 times in the drug-resistant Skbr3-LR cells (29.26 μmol/L). Compared with the parent cells

the expression level of the marker of epithelial cells

E-cadherin was decreased

while those of mesenchymal markers

Twist

Vinemtin

N-cadherin

and of YAP/TAZ were increased in the drug resistant cells (all

0.05). And Western blot assay also indicated the enhanced expression of YAP and p-YAP proteins (

0.05). After the Skbr3-LR cells were treated with verteporfin

the IC

to Lapatinib was decreased from 11.25 to 2.98 μmol/L

by 4 times

and there were less survival cells and the expression of YAP was decreased (

0.05).

Conclusion

The lapatinib-resistant model of breast cancer Skbr3-LR cells is successfully established. And the mechanism of cell resistant to lapatinib may be associated with the abnormal expression of YAP protein.

关键词

Keywords

references

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⁰

Role of yes-associated protein in resistance of HER2+ breast cancer to lapatinib

DOI：10.16016/j.1000-5404.201911189

摘要

Abstract

关键词

Keywords

references

Role of yes-associated protein in resistance of HER2⁺ breast cancer to lapatinib