Study On Effect And Mechanism Of Berberine Targetting Tubulin To Inhibit Proliferation In Epidermoid Carcinoma A431 Cell

Many pharmacological studies and a large number of clinical studies have confirmed that berberine has a significant anti-cancer effect.But target and the specific mechanism of its anti-cancer are rarely reported.The chemical structure of berberine is similar to that of colchicine,and the anti-cancer mechanism of colchicine is related to microtubules.Therefore,we use cell model experiments and transplanted animal model test to explore the specific mechanism of berberine resisting cancer whether is related to microtubules.First,molecular docking simulation experiments were performed to explore the binding sites of berberine and microtubules.And then the effect of berberine and microtubules was detected by biofilm interferometer and isothermal titration calorimeter.In this study,MTT assay was used to detect the effect of the survival rate of of berberine on eight kinds of tumor cell lines.After screening the appropriate cell lines,drug concentration and time,we treated cells with 10 ?g / ml of berberine for 24 h and 48 h,the status of microtubules in tumor cells was observed under inverted microscope.The expression of acetylated ?-tubulin was observed by western blotting.According to the results of in vitro experiments,the 6 to 8 weeks old male nude mice were divided into three groups according to the animal model of transplanted tumor.The control group(normal saline),cisplatin group(1mg / kg),berberine group(10mg / kg),each group has six nude mice,the weight of nude mice to monitor in nude mice.After the final nude mice were sacrificed,the liver,spleen and tumor were harvested were weighed,and the tumor was taken pictures and contrasted.It was found that berberine and microtubules were able to bind to the site of microtubule Gln8;In eight tumor cell lines,berberine had the best inhibitory effect on epidermal squamous cell carcinoma A431,and when the concentration of berberine was 2 ?g / ml,after treatment for 48 h,the inhibition rate of A431 cells reached more than 50%;berberine can disrupt the normal polymerization and depolymerization of microtubules in A431 tumor cells;berberine promotes the high expression of acetylated alpha-tubulin,which may promote microtubule stability;Comparing the berberine treatment group with the control group,there was no significant difference in the body weight,liver and spleen of the nude mice.The weight of the tumor and the size of the tumor volume had a decreasing trend,but there was no significant difference.In conclusion,we conclude that berberine targetting tubulin to inhibit proliferation in squamous cell carcinoma A431 cells,and almost has no side effects.This experimental conclusion opens up a broader field for the application of berberine in Chinese traditional medicine and also provides a reference for the study of the mechanism of similar related drugs.