Further Discussion Of The Mechanism Of Evodiamine Inducing Hela Cell Cycle Block By Building Synchronization Mode

This study was performed for the purpose of building a mode of Cell Synchronization of Hela, and applying this mode in the study of antitumour mechaniam of evodiamine。 Cell proliferation is a basic problem in biology. The period from the end of one cellular-fission to the end of the next cellular fission is called cell cycle. It includes interphase and mitosis phase, While interphase includes G1 stage, S stage and G2 stage. The cellular morphosis does not change much in interphase, but there is very active metabolism in this stage. Gl is DNA synthesis prophase;S is DNA synthesis phase and G2 is DNA synthesis anaphase in which tubulin is synthesized mostly and the synthesis of RNA and histone decreases. The biochemical changes in the different phase of cell cycle, actually, is a running process, The biosynthesis in prophase prepare the necessary material for the transition to the next phase, so if the biosynthesis in any phase of the cycle is blocked, the whole proliferation period will stop. Different cell phases have different sensitive chemotherapy drugs. We change the cells which are in different proliferation stage into the same stage of a cell cycle by artificial meas, that is called synchronization. Then giving some sensitive chemotherapy drugs according to different phase which can kill cancer cells more effectively as to cure tumor. 1, Cell Synchronization ExperimentAdd TDR solution to the culture with Hela cells in logarithmic phase in it, making the final concentration of TDR reach 2.0mol/L .Then the synthesization of DNA is restrained, cells in phase S stop, whle cells in otherphases go on until they stop at G/S, and that is The first interdiction. Remove TDR and wash the cells, then add fresh culture, and the cells get into new cycle. All cells escape S phase after 8h. Add TDR once more for The second interdiction, and after 15h, all cells are interdicted in a narrow zone of Gi/S. As a result, the proportion of G0/G1 is highest:95.27%, the highest proportion of G2/M is 87.39%, the highest proportion of S is 56.60%.2. Study of the mechanism of evodiamine inducing Cell Synchronization:Through the erperiment of evodiamine abducting unsynchro nizationd and Synchronizational cell ,we find that the cell blocked at G2.we got the conclution of that the cell Synchronization is benfit with the study of cell block. For example: someone investigating the evodiamine abducting unsynchRonization,the resault can not be fix on(G2 or S),Cell Synchronization mode is used in this text ,the cell blocking resault is G2 in evidence.Throught the experiment of evodiamine abducting unsynchronizationd and Synchronizational cell.we find that the cell blocking before the cell apoptosis.3. RT-PCR and Western Blot experiments of CyclinB 1We reach a conclusion through RT-PCR experiments of CyclinB 1 that " evodiamine hardly affects the transcription of cyclinBl, and that the expression of CyclinB 1 protein reduces. As we have demonstrated above that evodiamine can interdict Hela cells at G2 phase, we know that the expression of CyclinBl protein has something to do with the cells interdicted at G2phase. As to that the cell interdiction occur before apoptosis through the study before, the relation between CyclinB 1 and P53 will be our next study direction.