Development of anti-cancer drug targeted on cancer suppression gene products

点击翻译按钮获取中文摘要

Human breast cancer cell line.MCF-7, typical ER positive cell line, was subcultured and tested the following.Determination of NF2 protein in MCF-7 cell.NF2 protein in human breast cancer cell was identified by wetern blotting method. The anti-NF2 polyclonal antibody which commercially available was used for confirming existence of NF 2 protein. The NF2 protein which identified was about 78 kDa, the same molecular weight as reported before.Identification of NF2 protein expression by immunohistochemical staining in breast cancer tissue.Using anti-NF2 polyclonal antibody, immunohistochemical staining for the breast cancer tissue was performed. As a result, 25% of the cases were strongly NF2 positive. In cases of strongly positive staining, there were a high ratio of advanced disease, in contrast, in cases of negative staining, there were a lot of stage 1 disease.NF2 resolution by calpain, inhibition of decomposition by calpeptin.Same as the previous experiment, various density of calpeptin, which is the inhibitor of calpain, and other calpain inhibitors were added to the culture medium of MCF-7 cells and cell growth was observed. By western blotting, activation of the calpain was inhibited in dose dependent manner by calpeptin and by other calpain inhibitors. Inhibition of NF2 protein decomposition was also identified.Prediction of chemotherapeutic response by CD-DST method in human breast cancers.Preparing for the development of new anti-cancer drug, Collagen Gel Droplet Embedded Culture-Drug Sensitivity Test(CD-DST)in clinically available breast cancer tissues was pefarmed using highly effective drug, Cyclophosphamide+Epirubicin(CE)or Docetaxel, for breast cancer. As a result, concordance between CD-DST and clinical response of CE and DOC was 85.7% and 96.4%, respectively. It might be useful to perform CD-DST for the new drug under development.