†These authors contributed equally.
Academic Editors: Nicholas Pavlidis and Stergios Boussios
Background: Anlotinib, a multi-target tyrosine kinase inhibitor, has
significant anti-cancer effects on breast cancer (BC), lung cancer, colon cancer
and ovarian cancer, but its mechanism has not been investigated in BC.
Methods: The cell viability and growth of human non-triple negative BC
cell line MCF-7 and triple negative BC cell line MDA-MB-231 with the treatment of
anlotinib were tested by Cell Counting Kit-8 (CCK-8) assay and Ki67 staining. The
alteration of genes related to apoptosis and autophagy were investigated by
quantitative real-time reverse-transcription polymerase chain reaction (qRT-PCR),
western blots and immunocytochemistry (ICC). Cell apoptosis was valued by TUNEL
staining and flow cytometry. Further, mouse breast tumour cell lines AT-3 cells
were subcutaneously injected into C57BL/6 mice, and the effect of anlotinib
intragastrically on tumour growth in vivo was examined.
Results: We found that anlotinib suppressed the cell viability and
depressed Ki67 staining in MCF-7 and MDA-MB-231 cell lines. Besides, the drug
also enhanced cell autophagy and apoptosis of MCF-7 and MDA-MB-231 cell lines,
which could be rescued by autophagy inhibitors wortmannin (wort) and
3-methyladenine (3-MA), and BECN1 knockdown. Furthermore, Akt/GSK-3