| ObjectivesThe incidence of prostate cancer is significantly increased today,how to better control the growth and spread of cancer cells has become the top priority.Traditional Chinese medicine not only has extensive and profound cultural history,but also plays an important role in combating cancer.And rosmarinic acid in the fight against cancer application is very rich.Effects of rosmarinic acid on the biological behavior of human prostate cancer cell DU-145 were studied by means of cell-level detection of cytological changes.Methods1.Cultured prostate cancer cells DU-145,the concentration of rosmarinic acid was divided into 6 groups and blank control was set up.the effect of drugs on the survival and growth status of cancer cells was detected by using MTT method in the time period of 0,24,48 and 72,respectively.2.The cancer cells growing in logarithmic phase were treated with scratch,and the scratch cells were treated according to the dosing concentration of 70,140μM and the control group.12,24,48 h time points,respectively,phase difference microscope was used to observe the scratch width and scratch healing efficiency of the cells to judge the cell movement ability.3.Transwell three plates of cultured cells were selected in the experiment,one plate was the control group,the other two plates were added rosmarinic acid drug concentration was 70,140μM,after 48 h culture,the cells were collected,the cells after resuspension in the upper chamber,and the lower chamber was the medium.The basement membrane was not laid in the migration group,and the basement membrane was added to the invasion ability.After 48 h,the membrane cells were photographed.4.Divided three plates of cells into control group and 70μM concentration group and 140μM concentration group.after 48 h of rosmarinic acid action,flow cytometry was used to analyze the effect of rosmarinic acid treatment on the overall apoptosis DU-145 prostate cancer cells.5.Three cultured cells were divided into three groups:control group,rosmarinic acid 70μM concentration treatment group and 140μM concentration treatment group.the effect of rosmarinic acid on the cycle of prostate cancer cell DU-145 was detected by flow analyzer after 48 h culture.6.Two plates of cells were selected,one plate was set to control group,the other plate was treated with 140μM concentration of rosmarinic acid for 48 h to extract the protein,and each protein specific antibody was added.normal and phosphorylated P38,ERK,JNK protein expression in adenocarcinoma cell DU-145MAPK pathway was detected by Western Blotting method.Results1.MTT results showed that rosmarinic acid inhibited the growth of prostate cancer cell DU-145 with a concentration gradient of 135.8μM.at 48 time points.2.There was no significant change in cell motility after 12 h of rosmarinic acid,and when the action time was extended to 24 and 48 h,the DU-145 motility of prostate cancer cells decreased with the increase of rosmarinic acid concentration.3.The migration and invasion ability of prostate cancer cell DU-145 after 48 h treatment with drug rosmarinic acid decreased with the increase of drug concentration,and the cell migration and invasion ability was obviously weaker than 70μM concentration at 140 concentration.4.The DU-145 apoptosis of prostate cancer cells can be promoted after 48 h of rosmarinic acid treatment.the total number of apoptotic cells increases with the increase of rosmarinic acid concentration,and it is obviously more than 70μM concentration at 140 concentration.5.The DU-145 cycle of prostate cancer cells changed after 48 h of rosmarinic acid,the proportion of cells in different stages changed,the proportion of cells in G2/M and S stages under 140 concentration was less than 70pM concentration,and the proportion of cells in G0/G1 stage under 140 concentration was more than 70 concentration,indicating that rosmarinic acid action 48 h will block cells in the G0/G1 stage.6.The expression of p38 and JNK protein at 140μM concentration after 48 h of rosmarinic acid was significantly higher than that in control group,while the expression of ERK protein at 140 concentration was lower than that in control group,indicating that rosmarinic acid affected the amount and activity of phosphide protein expression.ConclusionRosmarinic acid can inhibit the survival rate of prostate cancer cell DU-145,slow down the motor ability of cells and promote the apoptosis of cancer cells in a certain concentration time range,and change the cell cycle.inhibit its migration and invasion ability,and the mechanism may be related to the regulation of MAPK pathways. |
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