[Development of three-dimensional breast cancer cell culture drug resistance model].
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.
Functional Laboratory, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
Hong
H
Xu
Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
Wei
W
Liu
Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
Xiu-Zhen
XZ
Zhang
Department of Cardiology, The Second Hospital of Dalian Medical University, Dalian 116027, China.
Liang
L
Hou
Department of Cardiology, The Second Hospital of Dalian Medical University, Dalian 116027, China.
Ying-Jin
YJ
Lu
Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
Pei-Pei
PP
Chen
Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
Can
C
Zhang
Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China.
Di
D
Feng
Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China. kongli@dmu.edu.cn.
Li
L
Kong
Department of Histoembryology, College of Basic Medical Sciences, Dalian Medical University, Dalian 116044, China. panpan1210@dicp.ac.cn.
Xiu-Li
XL
Wang
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D001943:Breast Neoplasms; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002470:Cell Survival; D004317:Doxorubicin; D019008:Drug Resistance, Neoplasm; D006801:Humans; D061986:MCF-7 Cells
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D004317:Doxorubicin