In Vitro Study On Mitomycin Induced Apoptosis In Body Cells Of Protoscolices

Objective: to study a phenomenon which mitomycin induced apoptosis on body cells of protoscolices in vitro. Methods: combing the glass pestle with 0.25% trypsin at 4℃ to digest protoscolices about 4~6 hours later, the body cells can be obtained. Using MTT to sieve a proper concentrations range of mitomycin, then, according to this range, the cells can be divide into 6 groups, and the concentration difference of the every group is 10μmol/L. The body cells of protoscolices were treated with different concentrations mitomycin for 24 hours and 48 hours. Detecting and observing the changes of cells under transmission electron microscope, agarose gel electrophotesis, fluorescent microscope and TUNEL method. Results: When incubated with different concentrations mitomycin for 24 hours, the body cells from 40μmol/L to 70μmol/L showed apoptosis under transmission electron microscope, chromatin condensation, even nuclear fragmentation, to form the apoptosis body can be seen. In 50μmol/L and 60μmol/L, the body cells expressed the obvious morphological changes associated with the characters of apoptosis under fluorescent microscope and TUNEL method. The number of apoptotic cells was the most in 60umol/L.After dying fluorescent, the percentage of apoptotic cells was 19.574%,compared with the control,p<0.01. However, typical nuclear fragmentation (DNA ladder) was not found during gel electrophoresis. Incubated with different concentrations mitomycin for 48 hours, the number of apoptotic cells was decreasing in every group,p<0.05, but the percentage of apoptotic cells in 60umol/L was still higher than other groups. Conclusion: found that mitomycin can induce apoptosis in body cells in vitro. Treated with 60umol/L mitomycin for 24 hours, the effect of apoptosis was the best.