Objective To study the effects and mechanisms of anlotinib on the proliferation, migration, invasion, and apoptosis of MYCN-amplified neuroblastomas (NB).

Methods  Two MYCN-amplified NB cell lines were treated with varying concentrations of anlotinib. CCK8 and clone formation assays were employed to detect cell proliferation, flow cytometry was used to detect cell apoptosis, scratch tests were performed to determine cell migration, and Transwell assays were used to detect cell invasion.

Results CCK8 assays indicated that anlotinib inhibited proliferation in MYCN-amplified NB cell lines in a concentration- and time-dependent manner. Clone formation assays showed that anlotinib significantly inhibited cell clone formation (P<0.000 1). The results of the apoptosis experiment showed that anlotinib significantly promoted apoptosis in MYCN-amplified NB cell lines (P<0.000 1). Scratch assays showed that anlotinib significantly reduced the migration ability of MYCN-amplified NB cells (P<0.000 1). Transwell assays revealed that anlotinib significantly inhibited the invasive abilities of MYCN-amplified NB cell lines (P<0.000 1). Western blot analysis demonstrated that anlotinib significantly reduced the phosphorylation of VEGFR2 and AKT proteins in MYCN-amplified NB cells. The expression of the key downstream proteins N-cadherin and Bcl-2 was significantly downregulated, whereas the expression of E-cadherin and Bax was significantly upregulated.

Conclusions Anlotinib markedly inhibits the proliferation, migration, and invasion of MYCN-amplified NB cells and induces apoptosis via the VEGFR2/PI3K/AKT signaling pathway.