KRAS inhibitor AMG510 combined with radiotherapy activates lung cancer immunity in an animal model
Objective To explore whether the combination of AMG510 and radiotherapy can enhance the therapeutic effect by enhancing anti-tumor immunity.Methods The sensitivity of the mouse Lewis lung cancer cell line(LLC)to AMG510 was detected using cell count kit 8(CCK8)in vitro,and the RAS gene sequencing was performed.Constructed a trans-plant tumor model using C57BL/6 mice and randomly divided them into 4 groups[control group,radiotherapy(RT)group,AMG510 group,and AMG510+RT group].They received treatment with AMG510 and/or radiotherapy respec-tively,and observed the inhibition of tumor growth.Detected the infiltration of T lymphocytes in tumors using flow cy-tometry and immunohistochemistry methods.Performed gene expression sequencing on the treated tumor and analyzed changes in immune related gene expression using methods such as heatmaps and signal pathway enrichment.The median test in one-way ANOVA or non parametric tests was used for inter group comparison.Results LLC cells had both KRAS G12C and NRAS Q61H mutations,and CCK8 showed that AMG510 alone inhibited the growth of LLC by 63％.In vivo experiments showed that AMG510 alone had limited effect on tumor growth,but the tumor growth volume was sig-nificantly controlled after combined radiotherapy,which was significantly better than any single therapy(all P＜0.05).Flow cytometry results showed that the proportion of tumor infiltrated CD3+T,CD4+T and CD8+T cells in the AMG510 combined with radiotherapy group was 3.29％(2.81％,6.44％),1.04％(0.72％,2.43％)and 2.16％(0.93％,4.19％),which was significantly increased compared with the other three groups(all P＜0.05),and the above results were consist-ent with the immunohistochemical results.In addition,immunohistochemical results showed that AMG510 combined with radiotherapy could increase the infiltration of interferon(IFN)-γ+T cells compared with control group and radiotherapy group(both P=0.009),but there was no statistical significance in the number of regulatory T cells(Treg)and macro-phage infiltration between the four groups,the x2 values were 2.22 and 0.50,respectively,and the P values were 0.528 and 0.918,respectively.Gene sequencing analysis showed that AMG510 combined with radiotherapy compared with the control group had the most differential gene data,including 399 up-regulated and 46 down-regulated genes,and the ex-pression of immune-related genes was up-regulated.AMG510 combined with radiotherapy and AMG510 enhanced chemo-kine and T cell receptor signaling pathways.Conclusion The combination of AMG510 and radiotherapy can significantly inhibit the growth of KRAS G12C mutant lung cancer and improve the anti-tumor immune function of T cells,which has translational medicine significance.
NETL
NSTL
万方数据
