The Cantharides Sodium Vitro Study, The Role Of Human Gastric Cancer And Esophageal Cancer Cells

Objectives:(1) To observe the effects of sodium cantharidinate on the growth of human gastric carcinoma cell line BGC823 and esophageal carcinoma cell line Eca109 at various concentration.(2) To study the effects of the sodium cantharidinate on cell cycle and cell apoptosis.(3) To study the expressions of the sodium cantharidinate on apoptosis related gene Bcl-2 and P53 in the tumor cells.Methods:(1) Cell and cell culture Human gastric carcinoma cell line BGC823 was gifted from the Department of Typhoid of Nanjing University of Traditional ChineseMedicine. Human esophageal carcinoma cell line Eca109 was purchased from Shanghai Cell Research Institute. All the cell lines were maintained in RPMI1640 containing 10% fetal calf serum and antibiotics at 37°C in a humidified atmosphere consisting of 5 % CO₂ Incubators . Once these cells had reached confluence, the cells were passaged by digesting with 0.25% trypsin.(2) Cell proliferation and inhibition test Cell proliferation and inhibition test was detected by MTT method. Human gastric carcinoma cell line BGC823 and esophageal carcinoma cell line Eca109 were harvested at exponential growth phase and digested with sufficient quantum 0.25% trypsin. These cell lines were prepared cell suspension with RPMI1640 containing 10% fetal calf serum and counted with blood cell plate. The cell suspensions were diluted with RPMI1640 and prepared 2 × 10⁵/ml respectively. The 180ul cell suspensions were plated in 96-well respectively. 20 μl physic liquor were added into cell suspensions in each pore, and every dose was repeated at four pores. Human gastric carcinoma cell line BGC823 and esophageal carcinoma cell line Eca109 were incubated at 37℃ in a incubator consisting of 5% CO₂ Incubators , and cell culture plates were taken out at 8h, 12h,24h,48h and 72h. MTT was prepared 1mg/ml at concentration with RPMI1640 free-of serum and added 100 ul into a pore and incubated 4h at 37℃, which deoxidized MTT into formazan.