Fig. 5

Aumolertinib exerts vascular normalization by decreasing VEGF secretion from EGFR-mutant tumor cells. A H1975 and HCC87 were cultured in serum-free media with or without the 2 μM aumolertinib and the VEGF amount in the media was measured by ELISA (n = 6). B Intratumoral amounts of VEGF amount in control group and Aumolertinib administration group were measured and compared (n = 8). C H1975 was pre-cultured for 24 h in the absence or presence of 2 μM aumolertinib, and then co-cultured with HUVEC following changing fresh medium. The migrations of HUVEC were monitored and images were shot at 16 h post co-culture. Meanwhile, the migrations of HUVEC were also monitored in the medium with or without aumolertinib to determine the direct influence of aumolertinib on HUVEC (n = 6). D Schematic presentation of the rebalance of angiogenesis and anti-angiogenesis factor in the tumor mass. Gene expressions of VEGF E and other mainly factors F in H1975 cell- and HCC827 cell-derived tumor xenografts with different drug treatment were assayed by qPCR (n = 8 for H1975 and n = 6 for HCC827). Gene expression was normalized to the housekeeping gene β-actin. G Endothelium and associated pericytes were visualized by CD31 (red) and α-SMA (green) immunofluorescence staining of H1975 and HCC827 tumor xenograft (n = 5). Representative images of different groups were shown. The fraction of CD31⁺ area and the relative proportion of α-SMA⁺ pericyte-covered blood vessels were quantified by image J. Con represents control; P represents pemetrexed; A represents aumolertinib; Osi represents osimertinib; P-A represents pemetrexed treatment followed by aumolertinib treatment. All of the data were expressed as the mean ± SEM, * p < 0.05, ** p < 0.01 and *** p < 0.001