ABSTRACT
Aurora kinase A (AURKA),a family member of aurora kinases,is involved in mitotic entry,maturation and separation of centrosome,assembly and stabilization of bipolar spindle,and condensation and separation of chromosome.Studies have demonstrated that AURKA plays a similar role in meiosis,while the specific mechanism and the similarities and differences in its role between meiosis and mitosis remain unclear.Therefore,we reviewed the studies about the localization and activation of AURKA in oocyte meiosis,and compared the role of AURKA in regulating spindle formation,activating spindle assembly checkpoint,and correcting the kinetochore-microtubule attachment between the meiosis of oocytes and the mitosis of somatic cells.This review will lay a theoretical foundation for revealing the mechanism of AURKA in the regulation of cell division and for the clinical research related to cancer and reproduction.
Subject(s)
Humans , Aurora Kinase A/genetics , Cell Cycle Proteins/genetics , Chromosome Segregation , Meiosis , OocytesABSTRACT
Objective@#To analyze the differentially expressed genes of patients with oral squamous cell carcinoma (OSCC) from paracarcinoma through biological information analysis to preliminarily identify OSCC-associated genes. @*Methods@#GSE23558, GSE37991 and GSE30784 were downloaded from the Gene Expression Omnibus (GEO), which is the mRNA expression profile dataset. The differentially expressed genes (DEGs) were identified based on the gene ontology and the Kyoto Encyclopedia of Genes and Genomes. Then, the protein-protein interaction (PPI) network was constructed using the STRING online tool, and Cytoscape was used to filter the critical genes. Furthermore, key genes involved in the survival of patients with OSCC were analyzed using Kaplan-Meier analysis. The expression of hub genes was validated based on GEPIA（http://gepia.cancer-pku.cn/）. @*Results @#A total of 212 DEGs were screened, and further analysis revealed 16 core genes, among which the core genes associated with prognosis included aurora kinase A (AURKA), aurora kinase B (AURKB), apoptosis inhibiting factor 5 (BIRC5), cell division cycle 6 (CDC6), E2F transcription factor 7 (E2F7), ubiquitin-like with PHD and ring finger domains 1 (UHRF1). These key genes were highly expressed in patients with oral squamous cell carcinoma, and the survival time of patients was short; the difference was statistically significant (P < 0.05).@*Conclusion @# AURKA, AURKB, BIRC5, CDC6, E2F7 and UHRF1 may be useful as potential biomarkers for OSCC prognosis prediction.
ABSTRACT
OBJECTIVE@#To investigate the effect of Aurora kinase B (AURKB) silencing-induced autophagy on apoptosis of osteosarcoma 143B cells and the underlying molecular mechanisms.@*METHODS@#Human osteosarcoma 143B cells were transfected with Lv/shAURKB or the negative control vector Lv/shScrambled followed by treatment with chloroquine (CQ) for 24 h. Western blotting was performed to detect the protein expression levels of AURKB, P62, LC3, cleaved caspase-3, Bcl-2, and P-ULK1. Transmission electron microscopy and LC3 dual-label fluorescence method were used to trace the autophagosomes in 143B cells to assess cell autophagy, and the cell apoptosis was detected using flow cytometry and TUNEL assay. Co-immunoprecipitation assay was used to detect the interaction between AURKB and ULK1.@*RESULTS@#The ratio of autophagy-related proteins LC3 II/I and the number of autophagosomes were significantly increased in 143B cells after transfection with Lv/shAURKB ( < 0.05), which significantly increased the expression of cleaved caspase-3 and reduced the expression of Bcl-2 ( < 0.05). Combined treatment of the cells with Lv/shAURKB and the autophagy inhibitor chloroquine obviously restored the expressions of caspase-3 and Bcl-2 ( < 0.05). Transfection with Lv/shAURKB significantly increased the apoptosis rate of 143B cells ( < 0.05), and this effect was significantly antagonized by combined treatment with chloroquine ( < 0.05). AURKB silencing strongly activated the phosphorylation of the autophagy-initiating protein ULK1 in 143B cells ( < 0.05). The results of co-immunoprecipitation assay confirmed when AURKB was immunoprecipitated, ULK1 also precipitated.@*CONCLUSIONS@#Silencing AURKB can induce autophagy by activating ULK1 phosphorylation to promote apoptosis in 143B cells.
ABSTRACT
OBJECTIVE In this research, the ubiquitin binding in ABIN and NEMO proteins (UBAN) domain of NF-kB essential modulator (NEMO) was used as a protein probe to screen potent linear ubiq-uitination substrates. Adopting the proteomics strategy, the putative substrates could be enriched and identified. This work will greatly expand our knowledge on the molecular mechanism of linear ubiquitina-tion regulation in various biological events. METHODS ® The glutathione S-transferase (GST)-UBAN protein was overexpressed by Escherichia coli and connected with glutathione agarose beads by GST tag to be used as the UBAN probe. Then, the function of the probe was verified in linear ubiquitination chain binding ability test by immunoprecipitation and Western blotting in 293T cells. Using the UBAN probe.linear ubiquitination substrates were enriched and identified by mass spectrometry in 293T cell lysate and analyzed by bioinformatics. Among them, the potential linear ubiquitination substrates Aurora kinase A (Aurora-A) and linear ubiquitin chain assembly complex (LUBAC) were co-transfected in 293T cells and the linear ubiquitination chain binding ability of Aurora-A was identified by Western blot. ® LUBAC was interfered in with siRNA and then synchronized in HeLa cells. The regulation of linear ubiquitination on the biological function of potential substrate Aurora A was observe by Western blotting. RESULTS (1) Western blotting results suggested that the UBAN probe could bind to the linear ubiquitin chain on NEMO, which confirmed the success of the system. ® Including Aurora-As a total of 403 proteins were identified as potential substrates of linear ubiquitination by mass spectrometry. The ability to bind to the linear ubiquitination chain was confirmed by immunoprecipitation between Aurora-A and the linear ubiquitination chains. ® Western blotting results showed that compared with the control group, Aurora-A phosphorylation level was up-regulated, which meant the occurrence of self-activation after knockdown of LUBAC. CONCLUSION The probe constructed with the UBAN domain can efficiently capture and enrich linear ubiquitination substrates. And the self-activation of Aurora-A may be regulated by linear ubiquitination. Significantly, the strategy of identifying the linear ubiquitination substrates established in this paper provides a novel idea and method for further exploration of biological functions of linear ubiquitination.
ABSTRACT
OBJECTIVE To explore the mechanism of Aurora kinase A (Aurora-A) promoting cancer cell chemotherapy resistance in nasopharyngeal carcinoma. METHODS The expression of Aurora-A in nasopharyngeal carcinoma tissues and adjacent tissues were detected by Western bolt and Q-PCR. The highexpressing Aurora A cell line CNE2 was used to detected the cell apoptosis and the expression of key pathway marker protein after Aurora-A inhibitor VX680 and cisplatin treatment by using Flow cytometry and WB. RESULTS The expression of Aurora-A in nasopharyngeal carcinoma tissues was significantly higher than that in adjacent tissues. Comparing to normal nasopharyngeal cells NP69, Aurora-A was significantly highly expressed in all of nasopharyngeal carcinoma cells and was highest in CNE2. Inhibiton of Aurora-A increased the cell apoptosis and the expression of p-AKT, p21 and Cleaved-Caspase-3 after using cisplatin or the Aurora-A inhibitor VX680 treatment. CONCLUSION The results shown that Aurora-A confer chemoresistance to cisplatin treatment through p-AKT/p21/Cleaved-Caspase-3 pathway.
ABSTRACT
OBJECTIVE@#To investigate the effect of danusertib (Danu), an inhibitor of Aurora kinase, on the proliferation, cell cycle, apoptosis, and autophagy of hepatocellular carcinoma HepG2 cells and explore the underlying mechanisms.@*METHODS@#MTT assay was used to examine the effect of Danu on the viability of HepG2 cells to determine the IC50 of Danu. The effect of Danu on cell cycle distribution, apoptosis and autophagy were determined using flow cytometry. Western blotting was used to detect the expressions of the proteins related to cell cycle, apoptosis and autophagy. Chloroquine was used to suppress Danuinduced autophagy to test the apoptosis-inducing effect of Danu.@*RESULTS@#Danu significantly inhibited the proliferation of HepG2 cells with IC of 39.4 μmol and 14.4 μmol at 24 h and 48 h, respectively. Danu caused cell cycle arrest in G/M phase in HepG2 cells and led to polyploidy accumulation via up-regulating the expressions of p53 and p21 and down-regulating the expressions of cyclin B1 and DC2. Danu also caused apoptosis of HepG2 cells through up-regulating the expressions of Bax, Puma, cleaved caspase-3, cleaved caspase-9, cleaved PARP and cytochrome C and down-regulating the expressions of Bcl-xl and Bcl-2. Danu induced autophagy via activating AMPK signaling and inhibiting PI3K/PTEN/AKT/mTOR axis, and inhibition of Danu-induced autophagy with chloroquine enhanced the pro-apoptotic effect of Danu.@*CONCLUSIONS@#Danu inhibits cell proliferation and induces cell cycle arrest in G/M phase, apoptosis and cytoprotective autophagy in HepG2 cells.
Subject(s)
Humans , Apoptosis , Autophagy , Benzamides , Pharmacology , Carcinoma, Hepatocellular , Pathology , Cell Cycle , Cell Division , Cell Proliferation , Hep G2 Cells , Liver Neoplasms , Pathology , Neoplasm Proteins , Metabolism , Protein Kinase Inhibitors , Pharmacology , Pyrazoles , PharmacologyABSTRACT
Objective: To investigate the effect of aurora kinase A (AURKA) activity on pro-angiogenesis, migration and invasion of Hep2 cells. Methods: Down-regulation of AURKA phospholation in Hep2 cells with 100 nmol/L VX680. Western blotting was used to detect the expression of p-AURKA, vascular endothelial growth factor A (VEGFA) and its receptor VEGFR2 in blank control Hep2 cells and Hep2 cells treated with VX680 for 24 h and 48 h. Angiogenesis experiment was applied to observe the effect of Hep2 cells treated with VX680 on angiogenesis. CCK8 cell proliferation assay, Transwell migration and invasion experiment were used to observe the ability of cell proliferation, migration and invasion of Hep2 cells. Results: Compared with control cells, the expression of p-AURKA in Hep2 cells treated with VX680 for 48 h was decreased (P=0.000). After downregulation of p-AURKA, the expressions of VEGFA and VEGFR2 were decreased (P=0.000). Meanwhile, angiogenesis was inhibited (P=0.000), and migration and invasion of Hep2 cells were reduced (P=0.000). Conclusion: AURKA regulates pro-angiogenesis, migration and invasion of Hep2 cells, which is a new strategy for the treatment of laryngeal cancer by targeting AURKA.
ABSTRACT
Objective To assess the expression level of targeting drug-based molecular biomarkers in ovarian clear cell carcinoma(OCCC)tissues and its clinical significance.Methods A total of 63 OCCC patients included 40 primary OCCC and 23 recurrent OCCC for secondary cytoreductive surgery(SCS),who had received primary surgeries at Fudan University Shanghai Cancer Center between January, 2008 and December, 2015 were enrolled, and immunohistochemistry SP method was used to test human epidermal growth factor receptor (EGFR), human epidermal growth factor receptor-2 (HER2), aurora kinase A (AURKA), breast cancer susceptibility gene 1 (BRCA1), BRCA2 and programmed death-ligand 1 (PD-L1) protein expression in paraffin-embedded tissues. Results The positive rates of EGFR, HER2, AURKA, BRCA1,BRCA2 and PD-L1 in primary and recurrent tumor tissues were respectively 20%(8/40)vs 30%(7/23),22%(9/40)vs 35%(8/23),38%(15/40)vs 35%(8/23),42%(17/40)vs 39%(9/23),20%(8/40)vs 22%(5/23), 25%(10/40)vs 17%(4/23), and there were no significant differences between primary and recurrent OCCC (all P>0.05). χ2-test or Fisher exact analysis revealed that HER2 expression in recurrent tumor tissues had a relationship with chemoresistance (P<0.05), while the expression of other biomarkers showed no significant relationship with chemoresistance (all P>0.05). Further, Kaplan-Meier survival analysis showed that patients with HER2 and AURKA-positive expression had a significantly shorter progression-free survival time in primary OCCC(4 months vs 10 months,log-rank test,P<0.05 for HER2;and 4 months vs 10 months,P<0.05 for AURKA);and a shorter overall survival time after SCS in recurrent OCCC (10 months vs 44 months, P<0.05 for HER2;and 13 months vs 43 months, P<0.05 for AURKA). However,multivariate Cox proportional hazards regression analysis indicated that none of these 6 biomarkers was independent risk factor of progression-free survival time of primary OCCC or overall survival time after SCS for recurrent OCCC (P>0.05). Conclusion HER2 and AURKA could serve as prognostic factors in ovarian clear cell carcinoma.
ABSTRACT
Coordination of cell division and cell fate is crucial for the successful development of mammalian early embryos. Aurora kinases are evolutionarily conserved serine/threonine kinases and key regulators of mitosis. Aurora kinase B (AurkB) is ubiquitously expressed while Aurora kinase C (AurkC) is specifically expressed in gametes and preimplantation embryos. We found that increasing AurkC level in one blastomere of the 2-cell embryo accelerated cell division and decreasing AurkC level slowed down mitosis. Changing AurkB level had the opposite effect. The kinase domains of AurkB and AurkC were responsible for their different ability to phosphorylate Histone H3 Serine 10 (H3S10P) and regulate metaphase timing. Using an Oct4-photoactivatable GFP fusion protein (Oct4-paGFP) and fluorescence decay after photoactivation assay, we found that AurkB overexpression reduced Oct4 retention in the nucleus. Finally, we show that blastomeres with higher AurkC level elevated pluripotency gene expression, which were inclined to enter the inner cell mass lineage and subsequently contributed to the embryo proper. Collectively, our results are the first demonstration that the activity of mitotic kinases can influence cell fate decisions in mammalian preimplantation embryos and have important implications to assisted reproduction.
Subject(s)
Animals , Mice , Aurora Kinase B , Metabolism , Aurora Kinase C , Metabolism , Blastocyst , Metabolism , Gene Expression Regulation, Developmental , Physiology , Histones , Metabolism , Phosphorylation , PhysiologyABSTRACT
BACKGROUND: Aurora kinase A (AURKA), or STK15/BTAK, is a member of the serine/threonine kinase family and plays important roles in mitosis and chromosome stability. This study investigated the clinical significance of AURKA expression in colorectal cancer patients in Korea. METHODS: AURKA protein expression was evaluated by immunohistochemistry in 151 patients with colorectal adenocarcinoma using tissue microarray blocks. We analyzed the relationship between clinicopathological characteristics and AURKA expression. In addition, the prognostic significance of various clinicopathological data for progression-free survival (PFS) was assessed. Also we evaluated copy number variations by array comparative genomic hybridization and AURKA gene amplification using fluorescence in situ hybridization in colorectal carcinoma tissues. RESULTS: AURKA gene amplification was found more frequently in the 20q13.2–13.33 gain-positive group than the group with no significant gain on the AURKA-containing locus. AURKA protein expression was detected in 45% of the cases (68/151). Positive staining for AURKA was observed more often in male patients (p = .035) and distally located tumors (p = .021). PFS was shorter in patients with AURKA expression compared to those with low-level AURKA expression (p < .001). Univariate analysis revealed that AURKA expression (p = .001), age (p = .034), lymphatic invasion (p = .001), perineural invasion (p = .002), and TNM stage (p = .013) significantly affected PFS. In a multivariate analysis of PFS, a Cox proportional hazard model confirmed that AURKA expression was an independent and significant prognostic factor in colorectal adenocarcinoma (hazard ratio, 3.944; p < .001). CONCLUSIONS: AURKA could serve as an independent factor to predict a poor prognosis in Korean colorectal adenocarcinoma patients.
Subject(s)
Humans , Male , Adenocarcinoma , Aurora Kinase A , Chromosomal Instability , Colorectal Neoplasms , Comparative Genomic Hybridization , Disease-Free Survival , Fluorescence , Gene Amplification , Immunohistochemistry , In Situ Hybridization , Korea , Mitosis , Multivariate Analysis , Phosphotransferases , Prognosis , Proportional Hazards ModelsABSTRACT
Aurora kinase A plays an essential role in mitosis including chromosome separation and cytokinesis. Aberrant expression and activity of Aurora kinase A is associated with numerous malignancies including colorectal cancer followed by poor prognosis. The aim of this study is to determine the inhibitory effects of LDD970, an indirubin derivative, on Aurora kinase A in HT29 colorectal cancer cells. In vitro kinase assay revealed that, LDD970 inhibited levels of activated Aurora kinase A (IC₅₀=0.37 mM). The inhibitory effects of LDD970 on Aurora kinase A, autophosphorylation and phosphorylation of histone H3 (Ser10), were confirmed by immunoblot analysis. Moreover, LDD970 inhibited migration of HT29 cells and upregulated apoptosis-related protein cleaved PARP. In cell viability assay, LDD970 was observed to suppress HT29 cell growth (GI₅₀=4.22 µM). Although further studies are required, results of the present study suggest that LDD970 provide a valuable insight into small molecule indirubin derivative for therapeutic potential in human colorectal cancer.
Subject(s)
Humans , Aurora Kinase A , Cell Survival , Colorectal Neoplasms , Cytokinesis , Histones , HT29 Cells , In Vitro Techniques , Mitosis , Phosphorylation , Phosphotransferases , PrognosisABSTRACT
This article was aimed to explore the effect ofSan-Huang (SH) decoction on improving chemosensitivity of MCF-7 cells to epirubicin and inhibition of Aurora kinase A, in order to discuss its underlying mechanism. The inhibition of MCF-7 cells proliferation on breast cancer by SH decoction was determined by CCK-8 assay. RT-PCR and western blot were used to detect the Aurora A, p53 mRNA and protein expression level of MCF-7 cells by SH decoction. The siRNA silenced Aurora A of MCF-7 cells. CCK-8 assay was used to detect the inhibition of MCF-7 cells proliferation. CCK-8 assay and AnnexinV-FITC/PI staining were used to detect the inhibition rate and apoptosis rate of MCF-7 cells treated by the combination of SH decoction and epirubicin. Western blot analysis was used to detect the expression of apoptosis-related proteins. The results showed that SH decoction inhibited the proliferation of MCF-7 cells in a dose-dependent manner (P 0.05). SH decoction can regulate the Aurora A, p53 protein and mRNA expression of MCF-7 cells. siRNA silenced Aurora A, which downregulated the inhibition rate of MCF-7 cells by SH decoction for 50.0% (from 49.2% to 24.8%). The combination of SH decoction and epirubicin enhanced the effect of epirubicin on inhibiting the proliferation rate and apoptosis rate of MCF-7 cell, regulated the expression levels of apoptosis-related protein such as c-PARP, c-Caspase 3, Bcl-2, Bax, as well as the protein level of Aurora A. It was concluded that SH decoction can increase the chemosensitivity of MCF-7 cells to epirubicin, which may be related to the inhibition of Aurora Kinase A by SH decoction.
ABSTRACT
BACKGROUND: The overexpression of Aurora A kinase (AurA) has been reported in various malignancies, including acute myeloid leukemia (AML). However, the expression of AurA and the effects of AurA inhibition in cancer stem cells are not yet fully understood. We investigated the expression and inhibition of AurA in AML stem cells (CD34+/CD38-). METHODS: Expression of AurA was investigated in cell lines (NB4 and KG1) that express high levels of CD34 and low levels of CD38. Primary AML cells were harvested from 8 patients. The expression of AurA and cell death induced by inhibition of AurA were analyzed in CD34+/CD38- cells. RESULTS: AurA was shown to be overexpressed in both primary AML cells and leukemia stem cells (LSCs) compared to normal hematopoietic stem cells. Inhibition of AurA plus cytarabine treatment in LSCs resulted in increased cytotoxicity compared to cytarabine treatment alone. Additional stimulation with granulocyte-colony stimulating factor (G-CSF) increased the cell death caused by AurA inhibition plus cytarabine treatment. CONCLUSION: To our knowledge, this is the first report describing increased expression of AurA in LSCs. Our results suggest that selective AurA inhibition may be used to reduce LSCs, and this reduction may be enhanced by stimulation with G-CSF. Further exploration of relationship between nuclear factor kappa-B and AurA inhibition and the potential of AurA inhibition for use in leukemia treatment is needed.
Subject(s)
Humans , Apoptosis , Cell Death , Cell Line , Cytarabine , Epilepsy , Granulocyte Colony-Stimulating Factor , Hematopoietic Stem Cells , Leukemia , Leukemia, Myeloid, Acute , Neoplastic Stem Cells , Phosphotransferases , Protein Serine-Threonine Kinases , Stem CellsABSTRACT
The Aurora kinase family of serine/threonine kinases plays an important role in chromosome alignment,segregation and cytokinesis during mitosis.Overexpression of Aurora kinases has been observed in a variety of human solid tumors and hematological malignancies.Aberrant expression of Aurora kinase can interrupt the function of the checkpoint in mitosis,lead to instability of genetic substance and induce tumorigenesis.
ABSTRACT
The abnormality of serine/threonine kinase Aurora-A is seen in many types of cancers. Although in physiological context it has been shown to play a vital role in cellular mitosis, how this oncogene contributes to tumorigenesis remains unclear. Here we demonstrate that Aurora-A overexpression enhances both the expression level and transcriptional activity of c-Myc. The inhibition of c-Myc expression by RNA interference significantly impaired the oncogenic potential of Aurora-A, resulting in attenuated cellular proliferation and transformation rates as well as fewer centrosomal aberrations. Furthermore, downregulation of c-Myc effectively overcame Aurora-A-induced resistance to cisplatin in esophageal cancer cells. Taken together, our results suggest an important role for c-Myc in mediating the oncogenic activity of Aurora-A, which may in turn allow for future targeting of c-Myc as a potential therapeutic strategy for tumors with Aurora-A overexpression.