Preliminary Study On The Effects Of Anlotinib On The Radiosensitivity Of Lung Adenocarcinoma H1299 Cells

Objective: As a small molecule TKI,Anlotinib has dual inhibitory effects on tumor cell growth and tumor angiogenesis.In this study,we aimed to explore the effects of Anlotinib on the radio-sensitivity of lung adenocarcinoma H1299 cells through in vitro experiments,so as to provide a basis for subsequent mechanism studies.Methods: Human lung adenocarcinoma H1299 cell line was cultured.CCK-8method was adopted to verify the proliferation toxicity of Anlotinib on H1299 cells,and IC20 concentration was calculated when the drug was used in subsequent experiments.The sublethal dose(6Gy was determined in previous studies of the research group)was prescribed.The experiment was divided into four groups,namely blank control group(NC),radiotherapy alone group(R),Anlotinib alone group(A),and Anlotinib + radiotherapy group(A+R).The cycle and apoptosis of H1299 cells in each group were detected by flow cytometry.Clonal formation experiment verified the effects of Anlotinib combined with radiotherapy on H1299 cells,calculated radiosensitivity-related parameters and radio-sensitization ratios,and plotted cell survival curve.Results:1.The results of CCK-8 cell experiment revealed different concentrations of Anlotinib(concentration: 0.01,0.1,1,10,and 100μmol/L)of lung adenocarcinoma H1299 cells.The results proved that Anlotinib could inhibit the activity of H1299 cells.With the increase of the concentration from 0.01μmol/L to 100μmol/L,the inhibition rate displayed an upward trend,and the survival rate illustrated a downward trend.The concentration of IC20 in H1299 cells was(0.387±0.274)μmol/L,and IC20 was used as subsequent experimental drug concentration.2.Flow cytometry demonstrated that the apoptosis rate in the A+R group was higher than that in the R group(P < 0.05).For H1299 cells,the percentage of S phase decreased and the percentage of G2/M phase increased in the A group compared with the NC group.Compared with the R group,the percentage of S stage,G2/M stage and G0/G1 stage decreased in the A+R group(P < 0.05),which further aggravated the damage effect of radiotherapy on lung adenocarcinoma H1299 cells.3.The results of cell clone formation experiment revealed that: by clicking the multitarget model and calculating the radio-sensitivity-related parameters,it was obvious that the Dq,D0 and SF2 of the A+R group were smaller than those of the R group,and the SER value was 2.052.The difference was statistically significant,indicating that Anlotinib had radio-sensitization effects on lung adenocarcinoma H1299 cells.Conclusion: Anlotinib can improve the radio-sensitivity of human lung adenocarcinoma H1299 cells,which affects the proliferation,cycle and apoptosis of tumor cells,and may provide a new option for the radio-sensitization therapy of NSCLC.