[Development of three-dimensional breast cancer cell culture drug resistance model.] [Article in Chinese]
Xu Hong, LIU Wei, ZHANG Xiu-Zhen, HOU Liang, LU Ying-Jin, CHENG Pei-Pei, ZHANG Can, FENG Di, KONG Li, WANG Xiu-Li
Functional Laboratory; Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China; Department of Cardiology, The Second Hospital of Dalian Medical University, Dalian 116027, China
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.
Key words: breast cancer; MCF-7 cell line; three-dimensional culture; drug resistance
Received: 2015-10-20 Accepted: 2015-11-17
Corresponding author: 孔力,王秀丽 E-mail: panpan1210@dicp.ac.cn, kongli@dmu.edu.cn
Citing This Article:
Xu Hong, LIU Wei, ZHANG Xiu-Zhen, HOU Liang, LU Ying-Jin, CHENG Pei-Pei, ZHANG Can, FENG Di, KONG Li, WANG Xiu-Li. [Development of three-dimensional breast cancer cell culture drug resistance model.] [Article in Chinese]. Acta Physiol Sin 2016; 68 (2): 179-184 (in Chinese with English abstract).