In Vitro Construction Of Three-Dimensional Culture Drug-Resistant Model Of Breast Cancer Cells
Keywords
Mcf-7, 3D Culture, Collagen Scaffold Material, Drug Resistance
Abstract
This study aims to construct a three-dimensional (3D) culture model of breast cancer in vitro, partially simulating the in vivo microenvironment of tumor cells, in order to establish an ideal breast cancer drug resistance research model. Breast cancer MCF-7 cells were combined with collagen scaffold materials to construct a 3D culture model of breast cancer in vitro. Carmine and HE staining were used to observe cell morphology, CCK-8 method was used to detect cell proliferation, live-dead cell staining kit was used to observe cell viability, and cell response to doxorubicin and doxorubicin under 3D culture and two-dimensional (2D) culture conditions was detected. Drug sensitivity to carboplatin and 5-fluorouracil, real-time quantitative RT-PCR to detect the mRNA expression levels of drug resistance-related genes P-glycoprotein (P-gp) and multidrug resistance-related protein 2 (mrp2), using Western blot detected the expression of P-gp protein. The results showed that MCF-7 cells in the 3D culture group grew in clusters, with good cell proliferation and activity. Compared with the 2D culture group, the sensitivity of cells in the 3D culture group to chemotherapy drugs was significantly reduced, the levels of P-gp and mrp2 mRNA were significantly increased, and the level of P-gp protein was increased to 5.3 times. The above results suggest that this study successfully constructed a 3D culture model of breast cancer with drug-resistant phenotype and function. This model is of great significance for further elucidating the mechanisms related to the occurrence of drug resistance.
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Original research was done by Hou Liang, Feng Di, Liu Wei, Kong Li, Zhang Can, Zhang Xiuzhen, Xu Hong, Wang Xiuli, Lu Yingjin, Chen Peipei
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