| Aim and Objective:Colon cancer is one of common malignant tumors in the digestive tract, with the continuous improvement of living standards, the incidence of colon cancer there is a rising trend. Later treatment of colon cancer is still present radiotherapy and chemotherapy based, but the poor quality of life of patients, as well as bone marrow suppression, allergic reactions and other adverse reactions, but a large number of clinical practice shows that the combination of Chinese and Western medicine, or cancer can improve efficacy, while can reduce the side effects of radiotherapy and chemotherapy. Some active ingredients of Chinese medicine can effectively inhibit tumor cell growth. Early studies have shown that evodiamine (EVO) induced apoptosis of tumor cells in G2/M arrest, there is M phase arrest (M-arrest) and M phase slippage (M-slippage) phenomenon.This study was jointly CDK1 inhibitors evodiamine CT26 mouse colon cancer cells, synergistic killing effect, based on the M phase block (M-arrest) and the M-slippage mechanism of sequential treatment is better than the same time dosing, and experimental methods and conditions of the previous basis from the pharmacodynamic aspects of tentative exploration and positive control evodiamine paclitaxel-induced M-arrest, add CDK1 inhibitors may be synergistic increase of apoptosis rate of the former to clarify tumors of this theory will provide a scientific basis for medicine, for the further search for more effective medicine for cell cycle components and develop mechanisms to drug targets to be the theory of traditional Chinese medicine prescriptions lay the foundation for development of new drugs against cancer medicine to provide experimental data. Methods:2.1 MTT method used to find evodiamine on CT26 cells induced irreversible point in time, the largest group with evodiamine 4mg/L, respectively, the role of time is 16h; 20h; 24h; 28h respectively after adding MTT, incubated for 4 hours, plus 150μl The DMSO solution, the Reader side, while another large group and the incubation time was maintained after the release of 12h, which with 10% fetal calf serum RPMI-1640 medium for 12h and then measured by MTT method MTT2.2 Paclitaxel and evoiamine combined with indirubin or R03306 sequentialadministration of both drugs on CT26 cell line. Evodiamine or paclitaxel group was 2mg/L,4mg/L group; R03306 (15mg/L) after dosing group or indirubin (20mg/L) after dosing (after 6 hours) group; R03306 or both indirubin dosing (30 hours) group; R03306 evodiamine and paclitaxel or sequential treatment group indirubin; R03306 evodiamine and paclitaxel, or both indirubin treatment group.2.3 Observed by colony formation assay and paclitaxel and evodiamine combined with R03306 or indirubin and sequential administration of both drugs inhibit the rate of CT26 cells.2.4 Comparison with flow cytometry evodiamine indirubin United R03306 or sequential treatment with both drugs on the CT26 cells; paclitaxel indirubin R03306 or sequential administration of both drugs and CT26 cell cycle, apoptosis index detection.Results:3.1 Evodiamine (EVO) induced apoptosis of CT26 cells irreversible time point, MTT results showed that the role of the first large group of evodiamine CT26 cells 16h; 20h; 24h; 28h after the inhibition rates were 42.1%±4.1%; 45.6%±11.1%; 47.6%±5.1%; 36.1%±5.6%; while another large group to maintain the culture time of 12h, after the release of inhibition rates were 35.0%±2.2%; 31.8%±4.0%; 58.9%±1.5%; 64.7%±4.2%. evodiamine 24h, time, EVO inhibition rate of release of the group significantly increased than the EVO; effect is greater than 24h, EVO EVO release group than in the group increased more significant inhibition (P<0.01), speculated that this When evodiamine induced apoptosis has entered an irreversible stage, irreversible evodiamine induced apoptosis may be a turning point of time of about 24h.3.2 Evodiamine (EVO) and paclitaxel combined with indirubin or CDK1 inhibitor R03306 or sequential administration of the synergistic killing effect of CT26, MTT results showed R03306 evodiamine and paclitaxel or sequential administration of indirubin significantly better than the synergistic anti-Evodia R03306 alkali and paclitaxel or the role of indirubin 30h while killing effect of drugs, the use of Kim Jong-q values are calculated, sequential administration of the q value is greater than 1.15, indicating synergy of synergies, while the q value of the treatment group were less than 0.85, no synergistic effect.3.3 Colony formation assay showed that Taxol and evodiamine the role of the United R03306 or CT26 cells of indirubin with sequential administration of both drugs inhibit the rate of CT26 cells, with the former group and drug, drug combination group showed sequential q-value greater than 1.15, for the synergies. While the q value of the treatment group were less than 0.85, no synergistic effect of 3.4. Flow cytometry showed the United CDK1 inhibitor evodiamine CT26 cells in a synergistic sequential drug, paclitaxel and the most specific CDK1 inhibitorConelusions:Evodiamine role of MTT at different time points were detected and after 24h, is evodiamine CT26 cells induced the turning point of this study, MTT, colony formation cloning method, flow cytometry studies and CDK1 specific inhibition evodiamine indirubin agent or combination R03306, R03306 paclitaxel and CDK1 inhibitor combination or indirubin, CT26 cells on the effects of colon cancer cells, the results show that:evodiamine and paclitaxel combined with R03306 or indirubin and sequential medication, can significantly improve the inhibition evodiamine rate of CT26, gaining synergies, but both are similar but no synergistic effect in combination. |
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