| The lack of the source tissue microenvironment in two-dimension (2D) culture led to monolayer cell growth, which was unable to form the approximate tumor tissue structure as in vivo. The significant differences of tumor cells structure between in vivo and in vitro led to the distinct differences of drug response pattern and the expression of signal molecular pathways. The long cycle, complicated operation, costly and individual differences of animal experiment, was not conductive to clinical application. The three-dimensional (3D) culture simulated the tumor micromicro-environment in vivo so that the structure of tumor cells in vitro and the expression of signaling pathways were approximate to the source tissue. In this study, we tried to establish a 3D culture model for prostate cancer, which could be utilized for drug sensitivity test in individualized tumor therapy.1. Established a 3D culture model based on Matrigel for prostate cancer cells. Analysis of the morphology of PC3 cells cultured in Matrigel, exhibited that PC3 cells formed tumor cell spheres with arborization-like gland tissue-like structure in Matrigel substrate, which was consistent with the PC3 cells in vivo. In addition, the proliferation kinetics of PC3 cells was determined and it displayed that growth of PC3 cells in Matrigel matrix rised steadily and kept active, which was similar to the growth pattern of PC-3 cells in vivo. Analysis of growth inhibition in PC3 cells by 13 anti-cancer drugs showed that the IC50 of 11 drugs was significant difference between 2D culture and 3D culture. PC3 cells in 3D culture was less sensitive to anticancer drugs than in 2D culture.2. Established a 3D culture model based on Alginate-HA for prostate cancer cells. This 3D microcarrier culture model provides basic research for future development of 3D cell culture for flowcytometer study. The cell morphology, the expression of tumor growth factors (IL-8 and VEGF), matrix metalloproteinases MMPs (MMP-2, MMP-3, MMP-9), biomarkers of EMT (N-Cadherin, Vimentin, β6 integrin) and the activity of apoptosis protein caspase-3 of prostate cancer cells in 2D cultures and 3D cultures based on Alginate-HA were Analysed. It showed that the expression of IL-8 and VEGF by prostate cancer cells in 3D culture was significantly up-regulated, meaning malignant phenotype significantly enhanced. In addition, the expression of MMPs and biomarkers of EMT by prostate cancer cells in Alginate-HA microcarrier greatly up-regulated, indicating remarkable increasement in invasion and metastasis potential, which was similar to that in vivo. The activity of caspase-3 of prostate cancer cells cultured in the 3D microcarrier significantly lower than that of 2D culture with anti-cancer drug treatment, which suggested that prostate cancer cells have obvious apoptosis tolerance in 3D culture based on Alginate-HA.3. Assessed the consistency of drug efficacy between in vivo and in vitro models. The activity of caspase-3 and Bcl-2 were compared among2D culture,3D culture based on Matrigel,3D culture based on Alginate-HA and nude-mouse transplanted tumor model with anti-cancer drug treatment. It showed that the apoptosis of two kinds of 3D culture was approximate to that in vivo, exhibiting obviously stronger apoptosis tolerability than 2D culture. It suggested that the drug response pattern of the two models of 3D cultures was similar to that in vivo. The 3D culture models can be further developed to replace animal experiments for antitumor drug efficacy evaluation. The Alginate-HA based 3D microcarrier is more cost-effective than 3D culture based on Matrigel, and may be further developed as 3D model for flowcytometer and microfluidity studies.Innovation:This is the first study using Alginate-HA combination to culture prostate cancer cells in 3D microcarrier; The consistency of drug response among 2D culture,3D culture based on Matrigel,3D culture based on Alginate-HA combination microcarrier and in vivo mouse model was evaluated for the first time in the present study. |
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