Anti-tumor Effects And Its Mechanisms Of 2-benzoylpyridine Thiosemicarbazone

Objective: To investigate the the protective anti-tumor effect and its Mechanisms of 2-benzoylpyridine thiosemicarbazone in k562 cells.Methods: With new compounds 2-benzoylpyridine thiosemicarbazone as the research object, Cisplatin as a positive control drug, observe the 2-benzoylpyridine thiosemicarbazone's proliferation effect in vitro on K562 cells through MTT assay,LDH leakage rate and the growth curve,observe the 2-benzoylpyridine thiosemicarbazone's proliferation effect in vitro on chang liver cells through MTT assay and LDH leakage rate,and investigate the anti-tumor activity and selectivity of 2-benzoylpyridine thiosemicarbazone in vitro. Detecting the apoptotic morphology in K562 cells through AO/EB staining by fluorescence microscope, detecting the mitochondrial membrane potential in K562 cells through Rh123 staining by fluorescence spectrophotometer,detecting the free intracellular calcium concentrationa in K562 cells through Fura-2/AM by fluorescence spectrophotometer,detecting the apoptosis in K562 cells through Hoechst33342/PI staining by HCS,investigate the mechanism of inducing tumor cell apoptosis of 2-benzoylpyridine thiosemicarbazone from the cell lever.Results: By MTT assay,after K562 cells treated with BT at different concentrations, the role of time is 24h,36h,48h, the results show that the proliferation of K562 cells are inhibited by BT,and the effect is showed the dose-dependent and time-dependent. Under the same concentration (50μM), BT is stronger than the role of CDDP. When acting on K562 cells 48h by BT, IC50 is 19.62μM. But the growth of Chang liver cells are hardly inhibited by BT. BT can cause the LDH leakage rate of K562 cells increasing,the growth of Chang liver cells almost are hardly inhibited by BT, LDH leakage rate is almost unchanged.The cell growth curve shows that the proliferation of K562 cells are inhibited by BT. From observing the morphological characteristics of K562 cells by AO/EB stainingthe the results show BT can promote apoptosis of K562 cells,cause the typical morphological features of apoptosis.Detecting the change s of MMP of K562 cells by Rh123 staining by the fluorescence spectrophotometer, the results show that BT can reduce MMP of K562 cells. Detecting fluorescence intensity under different wavelength spectrophotometer through the application of calcium-sensitive fluorescent probe Fura-2/AM by fluorescence spectrophotometer and calculating [Ca2+]I, the results show that, under the case of 1.3mmol/L extracellular Ca2+, [Ca2+]i was 219.2 nmol/L in normal K562 cells, [Ca2+]i in K562 cells increase in different degrees after K562 cells treated with BT.Determinating the apoptosis of K562 cells by Hoe-chest33342/PI staining,the results show that:the fluorescence intensities are increased following by increasing concentration of BT, the morphological pictures are showed the characteristics of apoptosis increase.Conclusions: The new compound BT can cause significant damage and apoptosis in K562 cells, and the effect is showed the dose-dependent and time-dependent.