The Research Of A New Copper Complex On Anti-tumor Effect Of And Its Mechanisms

Objective: To study a new type of copper complex (N-Cu) against tumor activity and its mechanism in vitro and in vivo.Methods: 1. The cytotoxicity of N-Cu for five human tumor (For example, SKOV3, Hela, MCF7, EC9706, A549) cell lines was tested by MTT in vitro.2. Mice with transplanted sarcoma (S180) Was injected N-Cu to celiac.The influence on mice were tested with weight, abdominal circumference, ascites volume, the rate of life extension and its toxicity on mouse main organs.3. Morphologic changes on Hela were observed by inverted microscope. The influence on Hela cell cycle and apoptosis of N-Cu were evaluated by Flow cytometry.4.Acting modes of N-Cu with Ct DNA was measured by using absorptions spectra, fluorescence spectra, Ct DNA thermal deformation experiment, Viscosity, gel electrophoresis tests . To explored Anti-tumor activity mechanism, Cleavage DNA test was performed.Results: 1. N-Cu inhibited the growth of the five human cell lines with different degrees. The 50% inhibitory concentration (IC₅₀) of N-Cu of five human tumor cell lines SKOV3, Hela, MCF7, EC9706 and A549 at 24 hours was 32.10±1.84, 168.39±3.24, 100.2±4.81, 49.05±1.91 and 24.35±2.76μg/ml, respectively. The 50% inhibitory concentration (IC₅₀) of N-Cu of five human tumor cell lines SK0V3, Hela, MCF7, EC9706 and A549 at 48 hours was 19.65±3.04, 15.7±0.14, 82.35±3.04, 33.2±1.98 and 21.65±2.62μg/ml, respectively.2. N-Cu showed a certain effect on transplanted S180. It decreased abdominal circumference and Ascites volume; It increased the rate of life extension on transplanted S180; It had no effect on main organs and the Liver and kidney of the treatment mice improved better.3. The results of Anti -tumor activity mechanism is as follows.(1) The result by cell cycle demonstrated that N-Cu could influence cell cycle of Hela. It reduced S phase and increased G₀/G₁ phase, and this effect showed dose dependence. All this proved that N-Cu could arrest cells in G₀/G₁ phase and induced cell apoptosis in the end.(2) N-Cu induced morphological changes of Hela cell. Compared with control group, N-Cu induced Hela cell apoptosis at the concentration of 10.00> 50.00,100.00μg/ml. The apoptotic rate was 3.19,11.01,24.10%, respectively.(3) Affected by N-Cu, the absorptions spectra of CT DNA appeared Hypochromic effect and Obvious red shift and the fluorescence spectra of DNA-EB generated weak fluorescence. The result by thermal deformation experiment: Tm arised Slightly. The result by gel electrophoresis displayed that N-Cu can transfer the ccc form of DNA in to oc and linear from. From all these We deduced N-Cu anti-tumor activity mechanism was related to the Injury of DNA.Conclusion: N-Cu have a good cytotoxicity of for five human tumor cell lines and biological activity in vivo. N-Cu could arrest cells in G₀/G₁ phase and induced cell apoptosis with high cell apoptosis rate. The result is the same with the Spectroscopy and the electrophoresis. So we can say N-Cu has a good development prospects for the anti-cancer drugs.