Growth and differentiation of carp maternal gene transformed human pituitary gland tumor cells

• Main Authors: Huang, Shiao-Wei, 黃曉薇
• Other Authors: Yew, Fu-Hsi
• Format: Others
• Language: zh-TW
• Published: 1993
• Online Access: http://ndltd.ncl.edu.tw/handle/59331900357990292828

碩士 === 國立臺灣大學 === 動物學系 === 81 === Human pituitary gland tumor cells (Hpt-1) were transfected with a maternal gene from carp egg. A transfectant cell line Hp-3 lost the ability to growth in soft agar. Immunoprecipitation and western blot analysis revealed that in Hp-3 the protein amount of epidermal growth factor receptor (EGFR) was considerably reduced as compared with the parental Hpt-1 cells, on the other hand that of glial fibrillary acidic protein (GFAP) and p53 were increased. Furthermore we found that Hp-3 cells could adhere and proliferate in bacteriological dish better than Hpt-1 cells, which suggested that Hp-3 cells secret a lot more of extracellular matrix into the medium. The result of western blot analysis indeed showed that the protein level of fibronectin of Hp-3 cell grown in bacteriological dishes was increased as compared with that in tissue culture dishes. A growth-stimulating activity was found in the serum- free medium conditioned by Hpt-1 or Hp-3 cells, which promoted the proliferation of hepatoma (NTU-BL), Hela, NIH 3T3 and human fibroblast cells except Hpt-1 and Hp-3 cells themselves. And there was no significant difference in the growth-stimulating activity between Hpt-1 and Hp-3 conditioned media. Hela and 3T3 were chosen as the main assay systems and exhibited a dose- dependence of mitotic response to the added conditioned medium. Since basic fibroblast growth factor (bFGF) has been examined as an autocrine regulator of glioma cell growth and a paracrine angiogenic factor facilitating its neoplastic progression, 0.5 ug /ml of anti-human bFGF antibody was added to the conditioned media of Hpt-1 and Hp-3 cells. However, the growth-stimulating activity was still not neutralized, suggesting that the mitogenic activity was not from bFGF.