title: Alisertib induces G2/M arrest, apoptosis, and autophagy via PI3K/Akt/mTOR- and p38 MAPK-mediated pathways in human glioblastoma cells.
abstract: Glioblastoma (GBM) is the most common brain tumor with poor response to current therapeutics. Alisertib (ALS), a second-generation selective Aurora kinase A (AURKA) inhibitor, has shown potent anticancer effects on solid tumors in animal studies. This study aimed to investigate the killing effect of ALS on GBM cell line DAOY and the possible underlying mechanisms using both bioinformatic and cell-based approaches. Our molecular docking showed that ALS preferentially bound AURKA over AURKB via hydrogen bond formation, charge interaction, and π-π stacking. ALS also bound key regulating proteins of cell cycle, apoptosis and autophagy, such as cyclin-dependent kinase 1 (CDK1/CDC2), CDK2, cyclin B1, p27 Kip1, p53, cytochrome C, cleaved caspase 3, Bax, Bcl-2, Bcl-xl, phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt), mammalian target of rapamycin (mTOR), 5'-adenosine monophosphate-activated protein kinase (AMPK), p38 mitogen-activated protein kinase (MAPK), beclin 1, phosphatase and tensin homolog (PTEN), and microtubule-associated protein light chain 3 (LC3). ALS exhibited potent growth-inhibitory, pro-apoptotic, and pro-autophagic effects on DAOY cells in a concentration-dependent manner. Notably, ALS remarkably induced G2/M arrest mainlyvia regulating the expression of CDK1/CDC2, CDK2, cyclin B1, p27 Kip1, and p53 in DAOY cells. ALS significantly induced the expression of mitochondria-mediated pro-apoptotic proteins such as Baxbut inhibited the expression of anti-apoptotic proteins such as Bcl-2 and Bcl-xl, with a significant increase in the release of cytochrome C and the activation of caspases 3 and 9. ALS also induced PI3K/Akt/mTOR and p38 MAPK signaling pathways while activating the AMPK signaling pathway. Taken together, these findings indicate that ALS exerts a potent inhibitory effect on cell proliferation and induces mitochondria-dependent apoptosis and autophagy with the involvement of PI3K/Akt/mTOR- and p38 MAPK-mediated signaling pathways in DAOY cells. ALS is a promising anticancer agent for GBM treatment.
authors.affiliation: Department of Neurosurgery, General Hospital of Ningxia Medical UniversityYinchuan, Ningxia, China; Ningxia Key Laboratory of Cerebrocranial Disease, Incubation Base of National Key Laboratory, Ningxia Medical UniversityYinchuan, Ningxia, China; Department of Pharmaceutical Sciences, College of Pharmacy, University of South FloridaTampa, FL, USA.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Neurosurgery, General Hospital of Ningxia Medical UniversityYinchuan, Ningxia, China; Ningxia Key Laboratory of Cerebrocranial Disease, Incubation Base of National Key Laboratory, Ningxia Medical UniversityYinchuan, Ningxia, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida Tampa, FL, USA.
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Zhou
authors.affiliation: Department of Neurosurgery, General Hospital of Ningxia Medical UniversityYinchuan, Ningxia, China; Ningxia Key Laboratory of Cerebrocranial Disease, Incubation Base of National Key Laboratory, Ningxia Medical UniversityYinchuan, Ningxia, China.
authors.forename: He-Chun
authors.initials: HC
authors.lastname: Xia
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of South FloridaTampa, FL, USA; Institute of Clinical Pharmacology, Department of Pharmacy, General Hospital of Ningxia Medical UniversityYinchuan, Ningxia, China.
authors.forename: Xin-Yu
authors.initials: XY
authors.lastname: Wang
authors.affiliation: Department of Colorectal Surgery, General Hospital of Ningxia Medical University Yinchuan, Ningxia, China.
authors.forename: Yin-Xue
authors.initials: YX
authors.lastname: Yang
authors.affiliation: Guizhou Provincial Key Laboratory for Regenerative Medicine, Stem Cell and Tissue Engineering Research Center & Sino-US Joint Laboratory for Medical Sciences, Laboratory Animal Center, Guizhou Medical University Guiyang, China.
authors.forename: Zhi-Xu
authors.initials: ZX
authors.lastname: He
authors.affiliation: Department of Neurosurgery, General Hospital of Ningxia Medical UniversityYinchuan, Ningxia, China; Ningxia Key Laboratory of Cerebrocranial Disease, Incubation Base of National Key Laboratory, Ningxia Medical UniversityYinchuan, Ningxia, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Sun
authors.affiliation: Department of Pharmaceutical Sciences, College of Pharmacy, University of South FloridaTampa, FL, USA; Department of Bioengineering and Biotechnology, College of Chemical Engineering, Huaqiao UniversityXiamen, Fujian, China.
authors.forename: Shu-Feng
authors.initials: SF
authors.lastname: Zhou
keywords: DAOY cell; Glioblastoma; PI3K/Akt/mTOR pathway; alisertib; apoptosis; aurora kinase A; aurora kinase B; autophagy; cell cycle; hydrogen bond; molecular docking