Development of three-dimensional breast cancer cell culture drug resistance model.
- Author:
Hong XU
1
;
Wei LIU
2
;
Xiu-Zhen ZHANG
2
;
Liang HOU
3
;
Ying-Jin LU
3
;
Pei-Pei CHEN
2
;
Can ZHANG
2
;
Di FENG
2
;
Li KONG
4
;
Xiu-Li WANG
5
Author Information
1. Functional Laboratory, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
2. Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
3. Department of Cardiology, The Second Hospital of Dalian Medical University, Dalian 116027, China.
4. Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China. kongli@dmu.edu.cn.
5. Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China. panpan1210@dicp.ac.cn.
- Publication Type:Journal Article
- MeSH:
ATP-Binding Cassette, Sub-Family B, Member 1;
Breast Neoplasms;
Cell Culture Techniques;
Cell Proliferation;
Cell Survival;
Doxorubicin;
Drug Resistance, Neoplasm;
Humans;
MCF-7 Cells
- From:
Acta Physiologica Sinica
2016;68(2):179-184
- CountryChina
- Language:Chinese
-
Abstract:
The aim of the present study was to develop three-dimensional (3D) culture model, a more pathologically relevant model, of human breast cancer for drug resistance study. MCF-7 cells were embedded within collagen gel to establish 3D culture model. Cellular morphology was observed using Carmine and HE staining. Cell proliferation was evaluated by CCK-8 assay, and cell activity was detected by Live/Dead staining kit. Drug sensitivities of the 3D culture to doxorubicin, carboplatin, 5-fluorouracil were assayed and compared with those of monolayer (2D) culture. In addition, the levels of drug resistance-related genes P-glycoprotein (P-gp), mrp2 mRNA expressions were detected by real time RT-PCR. Expression level of P-gp protein was detected by Western blot. The results showed that MCF-7 cells in 3D culture formed a number of cell aggregates, and most of them displayed good cell viability. The IC50 values of doxorubicin, carboplatin, 5-fluorouracil were all increased significantly in 3D culture compared with those in 2D culture. Moreover, compared with MCF-7 cells in 2D culture, the cells in 3D culture showed increased mRNA levels of P-gp and mrp2, as well as up-regulated protein expression of P-gp. These results suggest that in vitro collagen-embedded culture system of human breast cancer cells represents an improved pathologically relevant 3D microenvironment for breast cancer cells, providing a robust tool to explore the mechanism of drug resistance of cancer cells.
