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Mitosis is important for cell proliferation in eukaryotes, and chromosome replication and accurate separation are essential for cell division. Supervillin is a membrane and microfilament actin binding protein. Previous studies have found that it regulates the dynamic changes of the cortical distribution of F-actin and myosin II in cytokinesis, thus ensuring the correct distribution of the contraction ring and participating in the final completion of cytoplasm divisions. But it is not clear whether it functions during metaphase. Supervillin has several splicing isomers, among which supervillin isoform 4 (SV4) is the largest splicing isomer. In this study, the expression of SV4 in cells was reduced by the RNA interference method, and the dynamic process of mitosis and the morphology of astral spindles were detected and observed by real-time microscopy and immunofluorescence staining, and the potential molecular mechanism of SV4 in mitosis was analyzed. The results showed abnormal cell divisions after SV4 reduction: delayed transition from metaphase to anaphase (P<0.001), abnormal assembly of microtubules, a twofold-increase of the number of cells with multipolar spindles, and decreased γ-tubulin signaling in the centrosome (P<0.001). Through GST pull-down and mass spectrometry experiments, we found that SV4 and Aurora A bind to each other, and SV4 regulates the localization and activation of Aurora A in the centrosome during mitosis. In summary, supervillin plays an important role in mitosis. The isomer SV4 regulates spindle integrity and γ-tubulin recruitment by interacting with Aurora A and recruiting it for proper localization and activation in the centrosome during the metaphase, thus promoting the correct assembly of bipolar spindles and ensuring the accurate separation of chromosomes and the smooth progress of mitosis.
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Aurora kinase A (Aurora-A), a serine/threonine kinase, plays a pivotal role in various cellular processes, including mitotic entry, centrosome maturation and spindle formation. Overexpression or gene-amplification/mutation of Aurora-A kinase occurs in different types of cancer, including lung cancer, colorectal cancer, and breast cancer. Alteration of Aurora-A impacts multiple cancer hallmarks, especially, immortalization, energy metabolism, immune escape and cell death resistance which are involved in cancer progression and resistance. This review highlights the most recent advances in the oncogenic roles and related multiple cancer hallmarks of Aurora-A kinase-driving cancer therapy resistance, including chemoresistance (taxanes, cisplatin, cyclophosphamide), targeted therapy resistance (osimertinib, imatinib, sorafenib, etc.), endocrine therapy resistance (tamoxifen, fulvestrant) and radioresistance. Specifically, the mechanisms of Aurora-A kinase promote acquired resistance through modulating DNA damage repair, feedback activation bypass pathways, resistance to apoptosis, necroptosis and autophagy, metastasis, and stemness. Noticeably, our review also summarizes the promising synthetic lethality strategy for Aurora-A inhibitors in RB1, ARID1A and MYC gene mutation tumors, and potential synergistic strategy for mTOR, PAK1, MDM2, MEK inhibitors or PD-L1 antibodies combined with targeting Aurora-A kinase. In addition, we discuss the design and development of the novel class of Aurora-A inhibitors in precision medicine for cancer treatment.
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Chemotherapy is among the limited choices approved for the treatment of hepatocellular carcinoma (HCC) at intermediate and advanced stages. Preferential and prolonged drug exposure in diseased sites is required to maximize the therapeutic index of the drug. Here, we report an injectable supramolecular peptide hydrogel as an intraperitoneal depot for localized and sustained release of triptolide for the treatment of orthotopic HCC. We chose peptide amphiphile C-GNNQQNYKD-OH-based nanofibers as gelators and carriers for triptolide. Sustained triptolide release from the hydrogel was achieved over 14 days , with higher accumulation in and cytotoxicity against human HCC Bel-7402 in comparison with L-02 fetal hepatocytes. After intraperitoneal injection, the hydrogel showed prolonged retention over 13 days and preferential accumulation in the liver, realizing HCC growth inhibition by 99.7 ± 0.1% and animal median survival extension from 19 to 43 days, without causing noticeable pathological changes in the major organs. These results demonstrate that injectable peptide hydrogel can be a potential carrier for localized chemotherapy of HCC.
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Objective To investigate the sub-cellular distribution correlation between activated LIMK1 (pLIMK1Thr508) and Aurora-A in mouse oocyte meiosis,and changes in Aurora-A location and spindle structure in condition of LIMK1 inhibition.Methods Immunofluorescence staining was employed to detect the sub-cellular localization of pLIMK1Thr508and its spatial-temporal correlation with spindle organizing regulator Aurora-A in mouse oocyte meiosis; BMS-3, the specific inhibitor to LIMK1 activity, was applied to analyze the effects of LIMK1 inhibition on Aurora-A distribution and spindle formation. Results At meiotic prophase,pLIMK1Thr508was weakly detected and concentrated in the germinal vesicle(GV) in oocytes,with no signal of Aurora-A across the cytoplasm and nuclear area;as meiotic assumption approaching,pLIMK1Thr508left nuclear,aggregating as a single dense dote in the vicinity of nuclear, and being co-localized with the emerging Aurora-A; After germinal vesicle broke down (GVBD), pLIMK1Thr508and Aurora-A remained overlapped and concentrated as multi foci around the condensed chromosomes;at metaphase Ⅰ(MⅠ) and metaphase Ⅱ(MⅡ), pLIMK1Thr508was co-localized with Aurora-A on spindle poles;During anaphase Ⅰ(AⅠ) to telophase Ⅰ(Tel Ⅰ) progression, pLIMK1Thr508was detached from spindle poles and mainly concentrated on the cleavage furrow,while Aurora-A loosely congressed on spindle. In ad-dition, LIMK1 inhibition with BMS- 3 destroyed Aurora-A polar location and spindle formation. Conclusions pLIMK1Thr508is a microtubule organizing center (MTOC)-associated protein, may participate in spindle assembly and maintenance through regulating Aurora-A in mouse oocytes during meiotic progression.
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Aim To study the apoptosis effect of cucurbitacin Ⅱa on non-small cell lung cancer cell lines NCI-H460 and A549 and its underlying mechanism.Methods Cell viability was assessed by CCK-8 assay.The apoptosis effect and cell cycle arrest were detected by Flow cytometry.Western blot was employed to detect the related protein.Results The proliferation of lung cancer cell lines NCI-H460 and A549 was inhibited by CuⅡa, which showed cytotoxic activity with IC50 values of 224.9 nmol·L-1 and 108.3 nmol·L-1 against NCI-H460 and A549 respectively.CuⅡa induced the cells apoptosis and cell cycle arrest at G2/M phase.The results of Western blot showed CuⅡa inhibited the phosphorylation of STAT3 and Cofilin in a dose-dependent manner.Further, CuⅡa inhibited the phosphorylation of Aurora A, in line with the important characteristics of anti-tumor effect of Aurora A kinase inhibitor with blocking cells in the G2/M phase.Conclusion CuⅡa has obvious anti-tumor effect against non-small cell lung cancer, which suggests its value as a lead compound for lung cell carcinoma.
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Aurora A plays a key role in cellular mitosis. It is located in the centrosome and spindle, and is mainly involved in the processes of centrosome maturation and separation, bipolar spindle assembly, and the regulation of mitotic progression. Recent studies have suggested that Aurora A is involved in tumorigenesis and tumor development through multiple mechanisms. Overexpression of Aurora A could cause abnormal centrosome amplification, aneuploidy formation, and G2/M checkpoint defects, which result in chromosome instability and imbalance between cell division and apoptosis, and eventually leads to abnormal cell proliferation. Aurora A also participates in the regulation of the p53 and BRCA1 pathways, leading to suppressor gene dysfunction and changes in cell viability, and it induces telomerase activity by upregulating c-Myc, resulting in tumorigenesis. In addition, Aurora A also induces drug resistance in liver cancer cells. Thus, Aurora A has gradually become a new target for cancer therapy in recent years. This paper has summarized the recent studies on Aurora A, and reviewed its biological functions in cell mitosis and roles in liver tumorigenesis.
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Objective The objectives of this study were to investigate the expression of Aurora-A pro-tein in cervical cancer and precancerous lesions,and its relationship with human papilloma virus( HPV) infection, and to analyze the role of Aurora-A in the pathogenesis of cervical cancer. Methods One hundred cases of cer-vical biopsy or surgical resection specimens were collected from high-risk HPV( HR-HPV) test. There were 20 cases of normal cervical tissues,20 cases of CIN grade 1 ( CIN1 ) ,20 cases of CIN grade 2 ( CIN2 ) ,20 cases of CIN grade 3(CIN3),and 20 cases of cervical squamous cell carcinoma. The expression of Aurora-A protein was detected by immunohistochemistry and the correlation between Aurora-A expression and HR -HPV infection was analyzed. Results Aurora-A was highly expressed in cervical intraepithelial neoplasia and cervical cancer (P<0. 05),and its positive expression rate increased with the degree of cervical lesions. There was a positive correlation between Aurora-A expression and cervical cancer(r=0. 475,P<0. 001). There was a positive cor-relation between Aurora-A expression and HR-HPV infection in CIN2 and CIN3(V=0. 591,P<0. 05). Con-clusion Aurora-A may be associated with the development of cervical cancer. Aurora-A can be used as an important biomarker for the early diagnosis of cervical intraepithelial neoplasia or cervical cancer. It is also a po-tential therapeutic target for cervical cancer.
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Objective To investigate the effect of Aurora-A kinase inhibitors VX-680 on the proliferation of thyroid carcinoma cell and to study their molecular mechanism. Methods The undifferentiated thyroid carcinoma cells were divided into groups of VX-680 with different drug concentration. Drug effect was observed by MTT method to measure the changes of cell proliferation. The gene expression of Aurora-A , Bcl-2 and Bax were tested by immunofluorescence assay. Results MTT results showed cell proliferation had been suppressed obviously with the of concentration increase of drug and the duration of administration (P 0.05). Conclusions Aurora-A and Bcl-2 protein expression of thyroid cancer are inhibited when VX-680 being used , at the same time , Bcl-2/Bax scale value loses balance to accelerate forming Bax/Bax homodimer. Thus cancer cell proliferation is blocked and cell apoptosis is induced. So VX-680 is a good target inhibitor for anaplasia thyroid cancer.
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Purpose To explore the expression of Aurora kinase A (Aurora-A), minichromosome maintenance protein 7 (MCM7) and human papillomavirus type 16 E7 protein (HPV 16 E7) in uterine cervical squamous cell carcinoma (CSCC) and to investigate their relationship with clinicopathological factors. Methods Immunohistochemical method was employed on 20 cases of low-grade cervical intraepithelial neoplasia (CIN1) , 30 cases of high-grade cervical intraepithelial neoplasia (CIN2+3), 40 cases of CSCC, and 20 ca-ses of chronic cervicitis. Results (1) Aurora-A localized in the cytoplasm and nucleus. MCM7 protein positive staining localized in the nucleus. In the nucleus, and (or) the cytoplasm appeared brown particles positive for HPV 16 E7. (2) The expression of Aurora-A, MCM7 and HPV 16 E7 were higher in the group of CIN2+3 and CSCC than that in the group of chronic cervicitis or CIN1 ( P0. 05). Conclusion Aurora-A, MCM7 and HPV 16 E7 expression are gradually increased with disease progres-sion, and closely related to the occurrence and development of cervical cancer, they are expected to be early diagnosis, early treatment of biological indicators of cervical cancer.
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Objective Human lung adenocarcinoma SPC-A1/DTX cells have a higher radioresistance than SPC -A1cells. This study was to investigate the role of Aurora-An/uclear factor κB ( NF-κB) in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells and its possible molecular mechanisms . Metho ds We collected human lung adenocarcinoma SPC-A1 and SPC A1/DTX cells and divided them into four groups:sh-Aurora-A ( Aurora-A plasmid interference ) , sh-NC, NF-κB inhibition ( SPC-A1/DTX +NF-κB inhibitor ) , and DMSO control .We measured the in vitro radio-sensitivity of the cells by MTT assay , determined their proliferation ability by cloning assay , and detected the mRNA and protein expressions of the target genes by real -time quantitative RT-PCR and Western blot , respectively . Results The 50% effective doses ( ED50 ) of the SPC-A1 and SPC-A1/DTX cells on radiotherapy were (6.5 ±0.3) and (12.8 ±0.6) Gy, respectively, with statisti-cally significant difference between the two groups ( P <0.01 ) .In the radiation doses of 0, 2, 4, and 6 Gy, the numbers of the cloned SPC-A1 cells were 345 ±20 , 252 ±22 , 170 ±15 , and 81 ±10 , sig-nificantly lower than those of the cloned SPC -A1/DTX cells (402 ±21, 370 ±18, 301 ±16, and 252 ±15) (P<0.05).The protein and mRNA expressions of Aurora-A were remarkably higher in the SPC-A1/DTX than in the SPC-A1 cells (1.00 ±0.08 and 1.00 ±0. 06 vs 0.49 ±0.03 and 0.22 ±0.02, P<0.05).MTT assay showed a higher ED50 in the sh-NC than in the sh-Aurora-A cells ([11. 8 ±0.5] vs [7.1 ±0.3] Gy, P<0.01) as well as in the control than in the NF-κB inhibition group ([11.7 ±0.5] vs [6.1 ±0.3] Gy, P<0.01).Inhibition of Aurora-A increased the expression of IκBa by 2.18 ±0.32 times (P<0.01) and that of NF-κB by 0.24 ±0.03 times (P<0.01).The expressions of IκBa (1.00 ±0.05) and NF-κB (1.00 ±0.04) were significantly lower in the parent strains of SPC-A1 than 0.65 ±0.04 and 2.18 ±0.15 in the drug-resistant strains of SPC-A1/DTX (P<0.01). Conclusi on Auro-ra-A/NF-κB is involved in the radioresistance of human lung adenocarcinoma SPC-A1/DTX cells.
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Aurora-A is a mitotic serine/threonine kinase that is evolutionally conserved and localized at the centrosome. Aurora-A activation is required for mitotic entry, centrosome maturation, and centrosome separation. Aurora-A is frequently amplified and/or over-expressed in breast, ovarian, esophageal, colon, lung, and bladder cancers. Aurora-A has recently become a new target of malignant tumors. The Aurora-A inhibitor can be combined with other chemotherapeutic drugs to provide a new way for cancer treatment. In this study, we review the function and inhibitor of Aurora-A kinase.
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Objective:This study aimed to investigate the expression levels of centrosome-associated kinase Aurora-A, mutant type P53 (mt-P53), and c-myc in colorectal cancer. This study was also conducted to investigate the mutual relationship and functions of these factors in tumorigenesis and tumor progression. Methods:We examined the pathological specimens obtained from colorectal cancer, pericancerous tissues, and normal colorectal tissues by tissue microarray technique and immunohistochemistry (SP method) to determine the expression levels of Aurora-A, mt-P53, and c-myc proteins. The clinicopathological parameters were then analyzed. Re-sults: The positive rates of Aurora-A expression in normal colorectal tissues, pericancerous tissues, and colorectal cancer were 0%, 35%, and 69%respectively;by comparison, the positive rates of mt-P53 were 0%, 20%, and 57%, respectively. For c-myc, the positive rates were 0, 37%, and 76%, respectively. The expression levels of Aurora-A, mt-P53, and c-myc were significantly higher in tumor tis-sues than in normal colorectal tissues and pericancerous tissues (P<0.01). Aurora-A overexpression was related to the depth of invasion (P<0.05). Mt-P53 and c-myc overexpression was related to the depth of invasion, lymph node metastasis, and Dukes' classification (P<0.05). A strong positive correlation was observed between the expressions of Aurora-A, mt-P53, and c-myc in colorectal cancer (r=0.362, P<0.01; r=0.487, P<0.01). A strong positive correlation was also observed between the expressions of mt-P53 and c-myc in colorectal cancer (r=0.242, P<0.01). Conclusion:The overexpression of Aurora-A and c-myc and the mutation of P53 were important in tumorigenesis, tumor invasion, and metastasis of colorectal cancer. Thus, the co-detection of Aurora-A, mt-P53, and c-myc may be useful for the early diagnosis and prognosis of colorectal cancer.
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Background and purpose:Aurora-A is a member of serine/threonine kinase family. The abnormal expression of Aurora-A induces tumorigenesis and radioresistance. This study was aimed to investigate the association of Aurora-A with radioresistance. Methods: Capan-1 cells were treated with Aurora-A kinase inhibitor, and then used to test cell proliferation, anchorage independent assay, cell cycle, and cell cycle regulatory proteins. Treated cells were also used to detect cell apoptosis afterγ-irradiation. Results:Cell growth and colony number in soft agar were decreased after treatment with Aurora-A inhibitor. Treatment of cells with Aurora-A inhibitor also down-regulated the expression of Cyclin D1, CDK2 and CDK6 to induce cell cycle arrest at G1/S and G2/M phases, but promoted cell apoptosis afterγ-irradiation. Conclusion:Treatment of pancreatic cancer cells with Aurora-A kinase inhibitor blocks cell proliferation and cell cycle progression, and promotes sensitivity of cells to radiation. Thus, Aurora-A may be used as one of therapeutic targets to increase the sensitivity of pancreatic cancer radiotherapy.
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Objective To explore the expression of Aurora-A in breast carcinoma and the relationship with breast carcinoma treatment efficacy of paclitaxel. Methods The expression of Aurora-A were tested by immunohistochemical SP method in 67 cases of breast carcinoma. The relationship of Aurora-A with treatment efficacy of paclitaxel was analysed. Results The expression rate of Aurora-A was 73.13%(49/67)in breast carcinoma. The treatment effective rate of breast carcinoma was 77.78%(14/18) and the rate of nonresponder was 11.11%(2/18) in negative expression of Aurora-A;but the treatment effective rate was 34.69%(17/49) and the rate of nonresponder was 34.69%(17/49) in positive expression of Aurora-A. There were obvious correlations between expression of Aurora-A and treatment efficacy of paclitaxel. Statistical significance could be found between them (P = 0.013 ). Conclusion There is manifest correlation between the high expression of Aurora-A and resistance of paclitaxel in breast carcinoma.
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Objective To investigate the inhibitory effect of RNA interference on the expression of Aurora A in U251 cells, and the influence on proliferation and apoptosis of U251 cells. Methods The siRNA specific for Aurora A was synthesized and transfected into U251 cells in vitro. Aurora A mRNA expression and protein content were detected by RT-PCR and Western blotting respectively. The cell proliferation and apoptosis were observed by methyl thiazolyl tetrazolium(MTT) and flow cytometry(FCM). Transmission electron microscope was used to observe the ultrastructural changes of U251 cells. Results After transfection, the expression level of Aurora A mRNA was significantly decreased(P<0.01), and the protein content of Aurora A was also obviously reduced. The inhibitory rate of cell proliferation reached up to 67.57% 72 hours after transfection, which was significantly higer than that of normal control group(P<0.01). The apoptosis rate of U251 cells was significantly increased from (3.69±0.87)% to (15.34±2.16)% (P<0.01). Under the transmission electron microscope, it was observed that the U251 cells showed typical morphologic changes of apoptosis after transfection, such as karyopyknosis, chromatin condensation and margination, intracytoplasmic vacuoles formed, and apoptotic bodies formed. Conclusion The expression of Aurora A gene can be inhibited by siRNA successfully, and it results in the suppression of cell growth and induce apoptosis of human glioma cells in vitro. Aurora A may become a new target for the gene therapy of gliomas.
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Objective To detect the genetic amplification and protein expression characteristics of aurora-A in ovarian cancer and to interpret the role of aurora-A gene in course of onset,progression and regression phases of ovarian cancer.Methods The amplification of aurora-A gene was detected by quantitative PCR in 6 normal ovarian tissues and 8 ovarian cancer samples,and its protein expression was examined by immunohistochemistry in 6 normal ovarian tissues and 40 ovarian cancer samples,furthermore,the relationships between over-expression of aurora-A protein in ovarian cancer tissue and its pathologic classification,tissue differentiation,clinical phase,tumor proliferation trait and prognosis were analyzed.Results Quantitive PCR showed that aurora-A mRNA was significantly higher in 8 ovarian cancer samples than that in normal ovarian tissues(P0.05).Conclusion There are abnormal amplification and protein over-expression of aurora-A in ovarian cancer tissue,aurora-A probably play an important role in the onset and progression of ovarian cancer,and the novel biological treatment concerning aurora-A gene and its protein is probably a useful route for curing tumor.
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Objective To investigate the effects of Nordy on the proliferation,cell cycle,and the mRNA and protein expressions of Aurora-A in human ovarian cancer cell lines 3AO and SKOV3. Methods After being treated with Nordy at the doses of 25,50 or 100 ?mol/L,the proliferation of 3AO and SKOV3 cells were tested with MTT assay; The expression of Aurora-A was detected by RT-PCR and Western blotting; The effect on cell cycle were analyzed by flow cytometry ( FCM) . Results Treated with Nordy,the mRNA and protein level of Aurora-A gene were significantly reduced ( P
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Objective To compare and analyze the difference of ?-tubulin and aurora-A expression in human cervical cancer cells (CasKi ) and immortalized human cervical squamous H8 cells with positive HPV16 E6E7.Methods Difference of ?-tubulin and aurora-A expression in CasKi and H8 cells was analyzed by showing the fluorescence intensity of ?-tubulin with indirect immunofluorescence.Expression level of aurora-A mRNA was detected by RT-PCR.Expression level of ?-tubulin and aurora-A in CasKi and H8 cells was semi-quantitatively analyzed by Western blot.Results The immunofluorescence signal of ?-tubulin was stronger in Caski cells than in H8 cells (57.78?3.13 vs 37.37?2.37,P
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Objective:To investigate the inhibitory effect of RNA interference (RNAi) on the expression of Aurora-A in SKOV3 cells and on proliferation of SKOV3 cells.Methods:Two pairs of oligo small interference RNA (Oligo siRNA) specific for Aurora-A were designed for RNAi and were transferred into SKOV3 cells.The expression of Aurora-A were de- tected by RT-PCR and Western blot.Furthermore,the cell proliferation and apoptosis were observed by MTT and FCM af- ter transfection.Results:After transfection with Oligo siRNA,mRNA and protein level of Aurora-A gene in SKOV3 cells were obviously reduced,while the inhibitory rate of proliferation and apoptosis rate in SKOV3 cells were increased signifi- cantly.Conclusion:The Oligo siRNA specific for Aurora-A can reduce the expression of Aurora-A gene and induce apop- tosis of SKOV3 cells.