Inhibition of CSF1R and AKT by (±)-kusunokinin hinders breast cancer cell proliferation

• Published in: Biomedicine & pharmacotherapy 2020-09, Vol.129, p.110361-110361, Article 110361
• Main Authors: Rattanaburee, Thidarath, Tipmanee, Varomyalin, Tedasen, Aman, Thongpanchang, Tienthong, Graidist, Potchanapond
• Format: Article
• Language: English
• Subjects: AKT; Antineoplastic Agents, Phytogenic - chemistry; Antineoplastic Agents, Phytogenic - metabolism; Antineoplastic Agents, Phytogenic - pharmacology; Binding Sites; Breast cancer; Breast Neoplasms - drug therapy; Breast Neoplasms - enzymology; Breast Neoplasms - genetics; Breast Neoplasms - pathology; CDC2 Protein Kinase - genetics; CDC2 Protein Kinase - metabolism; Cell proliferation; Cell Proliferation - drug effects; CSF1R; Cyclin D1 - genetics; Cyclin D1 - metabolism; Female; Humans; Kusunokinin; Lignans - chemistry; Lignans - metabolism; Lignans - pharmacology; MCF-7 Cells; Molecular Docking Simulation; Molecular Dynamics Simulation; Protein Binding; Protein Conformation; Proto-Oncogene Proteins c-akt - metabolism; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor - antagonists & inhibitors; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor - chemistry; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor - genetics; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor - metabolism; Signal Transduction; Structure-Activity Relationship
• ISSN: 0753-3322; 1950-6007
• DOI: 10.1016/j.biopha.2020.110361

[Display omitted] •(−)-Kusunokinin bound CSF1R, MMP-12, HSP90-α, CyclinB1 and MEK1.•(−)-Kusunokinin bound CSF1R at juxtamembrane, sharing some P31 matching position.•(±)-Kusunokinin down-regulated CSF1R, AKT, CyclinD1 and CDK1 protein.•(±)-Kusunokinin inhibited cells proliferation partially through binding to CSF1R. Kusunokinin, a lignan compound, inhibits cancer cell proliferation and induces apoptosis; however, the role of kusunokinin is not fully understood. Here, we aimed to identify a target protein of (−)-kusunokinin and determine the protein levels of its downstream molecules. We found that (−)-kusunokinin bound 5 possible target proteins, including CSF1R, MMP-12, HSP90-α, CyclinB1 and MEK1 with ΔGbind less than −10.40 kcal/mol. MD simulation indicated (−)-kusunokinin and pexidartinib (P31, a specific CSF1R binding compound) shared some extents of functional similarity in which (−)-kusunokinin bound CSF1R at the juxtamembrane (JM) region with aromatic amino acids similar to pexidartinib using π-π interaction, as well as hydrogen bond. Both P31 and (−)-kusunokinin moved into the same CSF1R region and W7 was a mutual key residue. However, the P31 binding site differed from the (−)-kusunokinin binding site. For in vitro study, the synthetic (±)-kusunokinin exhibited stronger cytotoxicity than picropodophyllotoxin, silibinin and etoposide on MCF-7 cells and represented less toxicity than picropodophyllotoxin and doxorubicin on L-929 and MCF-12A cells. Knocking down CSF1R using a specific siRNA combination with (±)-kusunokinin demonstrated levels of cell proliferation proteins slightly higher than siRNA-CSF1R treatment. However, siRNA-CSF1R combination with P31 represented the number of cell viability and cell proliferation proteins, like in the control groups (Lipofectamine and siRNA-Luciferase). Moreover, (±)-kusunokinin suppressed CSF1R and its downstream proteins, including AKT, CyclinD1 and CDK1. Meanwhile, both P31 and siRNA-CSF1R dramatically suppressed CSF1R, MEK1, AKT, ERK, CyclinB1, CyclinD1 and CDK1. Our overall results indicate that the mechanism of (±)-kusunokinin differed fairly from P31. We have concluded that (±)-kusunokinin inhibited breast cancer cell proliferation partially through the binding and suppression of CSF1R, which consequently affected AKT and its downstream molecules.