| Background and objectiveThe researches on cancer inhibitor have been carried out for many years. Now, lots of materials have been identified to inhibit the growth of cancer. Except of the traditional chemotherapy medicine, Heparin, Hydrocortisone, Somatostatin and some NSAID can also inhibit the cancer. Recently, some Chinese medicines have been manifested to posses this ability: sodium cantharidate, sodium cantharidate, cinobufagin and coixenolide are reported frequently.Because of its favorable biocompatibility, Hydroxyapatite was extensively used in bone defect clinic as a substitute for bone. In 1992, Kano and his colleague prepared the hydroxyapatite crystallite by wet-hot method, and they did not sinter it to mass but prepared the crystallite as sol. The diameter of the hydroxyapatite crystallite is less than 100nm. At present, it is common to say that the material which crystallite diameter is less than 100nm is a nanometer material. The first generation of HAP-Sol was easy to congregate and deposit, so the posterity ameliorate the process to obtain steady stuff. In 1996, professor Li Shipu of Wuhan Industry University found that HAP-Sol was able to restrain some carcinoma cells, and he did primary research on it. He studied on the W-256 carcino-sarcoma cells, ascites cells, and A549 lung cancer cells successively by MTT method. However, not only the research on the cause or mechanism ofengendering such phenomenon is seldom, but also the works had been done existed some questions such as: whether the HAP-Sol influences the PH value of culture solution which leaded the cancer cells to death? There exists apoptosis in the process? Moreover, the effect of HAP-Sol on human tong cancer has not been investigated up to now. Consequently, our study plans to use the human tongue cancer cell Tca8113 as the object, and to investigate the effect of HAP-Sol on it We also arranged to test the apoptosis associated factor and proliferation activity of cancer cells to discuss the mechanism deeply.Materials and MethodsRPMI1640 culture solution was mixed with different concentrations of HAP-Sol, and the PH of the solution was tested. Using MTT colorimetric method, we can select the highest concentration of HAP-Sol to exclude the PH influence. Tca8113 cells were cultured with different concentrations of HAP-Sol in six-well plate, observing the cells at different time by phase-contrast microscope. Osteoblast in primary culture of rabbit was used as comparison; We harvest the cancer cells at different time, and dye with trypan blue, counting the number of cells and drawing the growth curve. Acid phosphatase was examined by Berry method which responses the activity of lysosome; The DNA of cells were extracted and identified by gel electrophoresis to observe the "DNA Ladder"; Dying with Hoechst33342 and observing the DNA breakage by fluorescence microscope; Using improved diphenylamine method to evaluate the DNA breakage rate; The ultrastructure of Tca8113 was examined by transmission electron microscope. In our study, blank RPMI1640 culture solution and common hydroxyapatite were used as negative comparison. After exoteric experiment, we established nude mice transplantation mode, and injected HAP-Sol directly into the mass. The tumor long and short diameter was measured every week, and counted the restrain rate; Nude mouse were sacrificedat the fifth week, and caspase3 immunochemistry dying and ACP dying were used to evaluatethe apoptosis of the mass.ResultsThe influence of PH could not be ignored when the HAP-Sol concentrations were up to 70ul, so the highest concentration in our study was 60ul/3ml culture solution. HAP-Sol could restrain Tca8113 obviously; this effect had dose dependency and time dependency. However, the effect on osteoblast could be neglected. It said that HAP-Sol could restrain cancer cells specifically. The cancer cells treated with HAP-Sol showed high activity of ACP which responses to lysosome. In experiment group, the DNA Ladder could be seen but not in osteobl...
|
PDF Full Text Request