Effects and mechanism of anlotinib on radiosensitivity of non-small cell lung cancer cell lines a549

Abstract

To investigate the effect and mechanism of Anlotinib on radio sensitization of human lung adenocarcinoma cell line A549. Human lung adeno carcinoma cell line A549 was teated with anlotinib and/or radiotherapy, then divided four groups, control group (Ctrl), Anlotinib treatment group (A), irradiation group (RT) and Anlotinib combined with irradiation group (A+RT) . CCK8 method was used to determine cell proliferation; the clone formation experiment was used to determine the inhibitory effect on cell growth; flow cytometry was used to determine cell cycle and apoptosis; immunofluorescence of γ-H2AX was used to determine DNA damage; expression of DNA-PKcs were detected by Western blot. Anlotinib inhibited proliferation and clonogenic survival following irradiation. The dose (Dq), the average lethal dose (D0) and the survival score (SF2) in the anlotinib combined radiotherapy group was significantly lower than those in the radiotherapy group. Anlotinib decreased G2/M phase arrest and promoted the cells apoptosis induced by in irradiation. The confocal microscopy results showed the average number of γ-H2AX foci in the A+RT group was more than that in RT group. The protein levels of DNA-PKcs were higher in A+RT group than that in RT group. Anlotinib enhances the radio sensitivity of A549 cells, which may be attributed to the delay DNA damage repair. It provides a rationale strategy by Anlotinib combined with irradiation for NSCLC.