Study On The Invasion And Migration Of Lung Cancer A549 Cells By X-ray Combined Brusatol

Objective: The objective of this study is to investigate the impact of brusatol in combination with 2Gy X-rays radiation on apoptosis,ROS,migration and invasion of lung cancer A549 cells,and to explore the potential molecular mechanism of transformation of tumor cells and brusatol combined with 2Gy X-rays.Methods: After brusatol titration and 2Gy X-rays radiation on lung cancer A549 cells,the rate of apoptosis and the change of ROS was assayed by flow cytometry;the migration rate of A549 cells was assayed by scratch test;invasion of A549 was assayed by Transwel l chambers;mRNA and protein of Nrf2,MMP-2,E-cadherin were detected by RT-qPCR and Western blotting.Results: The concentration of brusatol were inversely with the proliferation of A549 cells,with increase of concentration of brusatol,the proliferation became decreasing.The expression of Nrf2 was gradually decreasing and became more evident in 24 hours by utilization of brusatol in same concentration.This study selects 60 nM brusatol titration and 2Gy X ray radiation on A549 cells.The study were divided into control group,drug group,radiation group and combination group.The clone formation rate and detection of apoptosis results in control group were consistent with that in the drug group,radiation group and combined group.They can reduce the survival of the cell,inhibite the cell growth,and promopt the cell apoptosis,which was significantly more evident in combination therapy group compared with drug radiation group(p< 0.05).Brusatol could raise the sensitivity of A549 cells to X ray radiation.The test of ROS by flow cytometry demonstrated that the cells in fluorescence intensity area of drug and radiation group were much more than those in control group,and the cells in combination group were obviously move to the right side of fluorescence intensity area.The scratch test and the Transwell chamber test demonstrated that combination group could effectively reduce the migration and invasion of the cells.The real-time quantitative PCR and Western blotting test showed that both drug and combination group could significantly inhibit expression of Nrf2 and MMP-2 and up-regulate E-cadheri expression compared with control group(p<0.05).Conclusions: This study demonstrated that brusatol in combination with 2Gy X-rays irradiation on lung cancer A549 cells could increase the apoptosis of A549 and ROS content,reduce migration and invasion of the cells by regulate the expression of MMP-2 and E-cadherin,meanwhile we found that the clear relationship between MMP-2 and E-cadherin expression and Nrf2 expression.In conclusion,Brusatol,as a radiosensitizer,could promote apoptosis and inhibit the migration and invasion of A549 cells.Which provides data support for further laboratory research and theoretical basis for clinical application.