Stemness characteristics of nasopharyngeal carcinoma cells with cancer-associated fibroblasts in three-dimensional culture

• Main Authors: Ke-Yun Chyi, 綦可芸
• Other Authors: Yann-Jang Chen
• Format: Others
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/11550530614345840924

碩士 === 國立陽明大學 === 生命科學系暨基因體科學研究所 === 105 === Previous studies revealed that stemness properties of cancer cells are different whether they are in 2D or 3D culture environment. Microenvironment of cancer cells has important roles with the stemness of cancer. In addition, cancer-associated fibroblasts (CAFs) existing in the tumor microenvironment have better ability to promote tumor development than normal fibroblast. Therefore, we compared the influences on NPC stemness properties with different fibroblast in 3D culture system. NPC cell lines, TW01, TW06, Hone-1 and TW01 CSC, were used in this study. Fibroblasts were cultured from patients’ normal tissue or tumor tissue. Interleukin 6 (IL-6) and lactate were used to induce normal fibroblast to CAF and alpha-smooth muscle actin (α-SMA) and vimentin as the CAF markers were analyzed. Collagen type I was used as matrix to construct 3D culture environment. Expression of stemness markers, Oct-4 and Nanog, were assayed by Western blot. In 3D system, NPC cells could form more sphere structure than in 2D culture, that indicated the stemness of cells was increased in 3D system. Nanog expressed more in 3D mono-culture than 2D system by western blot. However, Nanog was decreased in cancer stem cell (CSC) of NPC. In the other side, CAF markers were increased after treated with IL-6 or lactate in normal fibroblasts, as the previous research. When NPC cells were co-cultured with CAF, the protein level of Nanog was increased. In addition, Oct-4 and Nanog both enhanced, when CSC co-cultured with fibroblast treated with lactate. Furthermore, the concentration of lactate which released to medium rose if NPC cells/CSC co-cultured with CAF. In this study, we found that 3D culture environment with fibroblasts might change stemness properties of cancer cells.