Effects Of Novel CDK7 Inhibitor On The Biological Function Of Human Glioma Cells

Objective To study the effects of a new CDK7 inhibitor THZ1 on the proliferation,apoptosis,invasion and cycle of human glioma U87 and U251 cells in vitro.Method The U87 and U251 cells were divided into blank group,control group and experimental group.The blank group was added with DMEM complete culture medium.The control grouq)was added with DMEM complete culture medium containing 1/1000 DMSO(sulfoxide dimethyl).The experimental group was treated with different concentrations of THZ1.The inhibitory effect of THZ1 on cell proliferation was examined by CCK-8(Cell Counting kit 8)and colony formation assay.The invasion ability was evaluated by Transwell assay.The cell cycle distribution and apoptosis rate of U87 and U251 cells induced by THZ1 were detected by flow cytometry(FCM).The expression of apoptosis related protein Caspase-3 was detected by blot Western assay.Result 1.THZl inhibits the proliferation of U87 and U251:After treatment in each group for 72 hours,there was no significant difference between the blank group and the control group(P>0.05).each concentration of experimental group was able to inhibit the proliferation of cells,and with the increase of THZ1 concentration,the inhibition degree is more obvious.According to the experimental results,SPSS20.0 analysis was used to analyze the drug IC50(half maximal inhibitory concentration),and IC50 of U87 cells were 83.1nmol/L and U251 cells were 13.7nmol/L.The plate clone formation test results show that even very low concentration of drug could inhibit colony formation rate2.Glioma cells U87 has a strong ability to invasive,In vitro transwell invasion experiments showed that the number of cells in the control group was 134 ± 13,the number of cells in the 50nmol/L treatment group was 72 ± 7,After 24 hours of treatment with 50nmol/L THZ1,the invasive ability of the control group was significantly lower than that of the control group(t = 7.141,P<0.01).Therefore,THZ1 can significantly reduce the invasiveness of glioma cells.3.THZ1 promoted the apoptosis of U251 cells and increased the expression of Caspase-3.The apoptotic rate was(8.3 ± 2.5)%in the control group,(15.6 ± 3.7)%in the 10 nmol/L group,(39.7 ± 6.0)%in the 20 nmol/L group,The apoptotic rate increased with the increase of THZ1 concentration.Compared with the control group,the difference was statistically significant(t = 2.832 and 8.367,respectively,P<0.05).Western blot analysis showed that the expression of Caspase-3 was significantly increased with the increase of THZ1 concentration.4.THZ1 block U87 and U251 cell cycle progression:the proportion of G2 phase cells increased significantly,compared with the control group,the difference was statistically significant(P<0.05)Conclusion THZ1 can inhibit the proliferation,promote apoptosis,inhibit the ability of invasion and block cell cycle progression of U87 and U251 in glioma cells.