| ObjectiveCurcumin is the major effective component of curcuma, which is a kind of tradition Chinese medicine. It has been paid more attention to curcumin recently for its specific function of anti - oxidation , anti - inflammation ,lipid - regulation ,anti - thrombin,anti - arteriosclerosis and anti - tumor. The study about its anti - tumor function has already become the highlight of today. For its broad -spectrum of anti - tumor ,little side effect and low price, curcumin is a new kind of anti - tumor drug with bright future. The study concerning curcumin anti -tumor effect is mainly focused on tumor chemical prevention, and America national tumor institute has ranked it as the third generation drug for cancer chemical prevention. Present studies suggest curcumin has the following functions: 1. anti - nitrolysation; 2. anti - NO; 3. anti - synthesization of DNA 4. anti - oxidation damage of DNA; 5. anti - vessel generation; 6. inducing tumor cell differentiation ; but the exact mechanism of anti — tumor is not clear. This study is designed to investigate the possible anti - tumor mechanism and molecular mechanism of curcumin through the trial in vitro in human colon cancer CX -1 cells.MethodsHuman colon cancer cell group CX -1 was sustained in the RPMI1640 culture medium containing 10% serum of fetal calf , glutamine , penicillin and streptomycin, and cultured in the box with 5% CO2 and saturated moisture. When cells developed into logarithmic growth stage, the experiment was performed. The inhibition of cells growth induced by various concentration of curcumin indifferent course was analyzed by using MIT assay, at the same time experiment group and control group were both set, then using machine to detect light - absorption rate at wave 570nm. Inhibitory rate of tumor cell ( % ) = (1 - A value of experiment group /A value of negative control group) 100% ; Certain concentration curcumin was processed till the schedule time when the cells in the culture medium were collected, and made into cell — suspending fluid with concentration of 1 107/L. The nuclei were stained by acridine orange fluorescent staining method and the morphological changes of the cell nuclei were examined under fluorescence microscope; development,morphological changes and apop-totic process of CX -1 cells for both control and experiment groups were detected under light microscope; Super - microstructure of CX - 1 cells were observed under transmission electron microscope, especially for the apoptotic structure of the cells in experimental group; The cell apoptosis ratio of CX - 1 processed with curcumin in different times were measured by flow cytometry a-nalysis and compared with control group. It has already revealed that there is an relationship between the processing time of curcumin and the apoptosis rate; The quantity of apoptosis - inhibiting protein Bel - 2 and apoptosis - promoting protein bax were evaluated by immuno - histochemistiy method both before and after processed with certain concentration curcumin; positive and negative contrast were set in the trial.ResultsGrowth inhibitory rates of CX - 1 cells by curcumin were significantly different in dose - dependent manners. Human colon cancer CX - 1 cells were processed respectively with 1,2,4,8,16,32ug/ml curcumin and 72 hours later the inhibitory rates revealed to be 20.67% ,40.97% ,67.75% ,83.05% ,85.28% , 91.24% , then compared with controlled group P <0.01; Chromatin condensation and nuclear fragmentation were seen under fluorescence microscope in cells treated with acridine orange fluorescent staining method. The apoptotic cells were observed under transmission electron microscope and elected by flow cytometry. Another apex of sub - diploid appeared in front of G0/G1 apex, and... |
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