Research On The Interaction Of Nano-hydroxyapatite With Ribosome

The inhibition effect of Nano-hydroxyapatite on cancer cell proliferation is a hot issue in the field of biological research of inorganic nanoparticles. The thesis focused on the interaction of nano-hydroxyapatite with the ribosome, and aimed to further illustrate the tumor suppressor mechanisms of nano-hydroxyapatit. This paper mainly studied the tumor suppressor role of nano-hydroxyapatite through the following aspects:the effects of nano-hydroxyapatite on cancer cell and normal cell proliferation; the effect of nano-hydroxyapatite on cell protein synthesis; the binding status of nano-hydroxyapatite with protein, DNA and RNA; the binding status of nano-hydroxyapatite with ribosomes.The hydroxyapatite nano-particles were prepared by chemical deposition method. Its effects on normal cells and cancer cells proliferation were studied using the Alamar Blue method and MTT assay separately. The results showed that the roles of nano-hydroxyapatite on cancer cells and normal cell proliferation were significant different. Nano-hydroxyapatite inhibited the proliferation of cancer cells in a dose-dependent maner, while had no significant inhibitory effect on normal cells. After treating cells with hydroxyapatite, the total RNA was extracted from the cells and the protein synthesis levels were measured using a cell-free expression system. It was validated that the nano-hydroxyapatite inhibited cancer cell proliferation at protein level. Using spectrophotometry, the binding status of nano-hydroxyapatite with protein, DNA, RNA was studied. It was found that the nano-hydroxyapatite with different sizes poessed different combination capacity with bovine serum albumin, pUC18, and the rat liver total RNA. Finally, the binding status of nano-hydroxyapatite with ribosome was studied. The rat liver ribosomes were purified and incubated with nano-hydroxyapatite in vitro, then the binding status of mRNA and ribosome was studied using flurosecence intensity analysis and SDS-PAGE. The results showed that nano-hydroxyapatite binded strongly to ribosomes, thus inhibited mRNA binding to ribosomes.With a series of experiments, this study confirmed that nano-hydroxyapatite can interact with cell protein, DNA, RNA, bind to ribosome, disturb the interaction of mRNA with ribosomes, thereby disrupt the protein synthesis, and ultimately, significantly inhibit the proliferation of cancer cell.