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Man Tong - One of the best experts on this subject based on the ideXlab platform.
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Abstract 901: Antibody-mediated blockade of Annexin A3 (ANXA3) synergistically enhances the efficacy of sorafenib and regorafenib for hepatocellular carcinoma
Experimental and Molecular Therapeutics, 2018Co-Authors: Man Tong, Lei Zhou, Noelia Che, Phillis Kau, Steve T. Luk, Xin Chen, Jin Ding, Terence K. LeeAbstract:Advanced hepatocellular carcinoma (HCC) is a lethal malignancy with limited treatment options. Sorafenib is the only FDA approved first-line targeted drug for advanced HCC, but its effect on patients9 survival gain is limited and patients ultimately present disease progression. Molecular mechanism underlying acquired resistance are still far from being well understood. A better understanding of causes of sorafenib resistance, enhancing the efficacy of sorafenib and finding a reliable predictive biomarker are crucial to achieve efficient control of HCC. In this study, we uncovered a role of Annexin A3 (ANXA3) in conferring ability of HCC cells to resist sorafenib. ANXA3 was found highly enriched in sorafenib-resistant HCC cells and patient-derived xenografts. Mechanistically, we found overexpression of ANXA3 in sorafenib-resistant HCC cells to suppress PKCδ/p38 associated apoptosis and activate autophagy for cell survival. ANXA3 expression correlated positively with LC3B, an indicator of autophagy induction, in HCC specimens and was associated with a worst overall survival in HCC patients who went on to receive sorafenib treatment. Anti-ANXA3 therapy, alone or combined with sorafenib and regorafenib, the standard of care for advanced HCC, impaired tumor growth in vivo and significantly increased survival. In sum, we found anti-ANXA3 therapy to exhibit abilities to reduce tumor growth and sensitize HCC cells to sorafenib and regorafenib treatment via suppressing autophagy and activating apoptosis. Inhibition of ANXA3 would benefit a subset of patients after sorafenib progression. ANXA3 can also serve as a useful predictive biomarker for patient response to sorafenib treatment. Citation Format: Man Tong, Noelia Che, Lei Zhou, Phillis W. Kau, Steve T. Luk, Xin Chen, Jin Ding, Terence K. Lee, Stephanie Ma. Antibody-mediated blockade of Annexin A3 (ANXA3) synergistically enhances the efficacy of sorafenib and regorafenib for hepatocellular carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 901.
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Efficacy of Annexin A3 blockade in sensitizing hepatocellular carcinoma to sorafenib and regorafenib.
Journal of hepatology, 2018Co-Authors: Man Tong, Lei Zhou, Noelia Che, Steve T. Luk, Phillis W.f. Kau, Stella Chai, Elly Sau-wai Ngan, Kwan Man, Jin DingAbstract:Background & Aims Advanced hepatocellular carcinoma (HCC) is a lethal malignancy with limited treatment options. Sorafenib is the only FDA-approved first-line targeted drug for advanced HCC, but its effect on patient survival is limited. Further, patients ultimately present with disease progression. A better understanding of the causes of sorafenib resistance, enhancing the efficacy of sorafenib and finding a reliable predictive biomarker are crucial to achieve efficient control of HCC. Methods The functional effects of ANXA3 in conferring sorafenib resistance to HCC cells were analyzed in apoptotic and tumorigenicity assays. The role of ANXA3/PKCδ-mediated p38 signaling, and subsequently altered autophagic and apoptotic events, was assessed by immunoprecipitation, immunoblotting, immunofluorescence and transmission electron microscopy assays. The prognostic value of ANXA3 in predicting response to sorafenib was evaluated by immunohistochemistry. The therapeutic value of targeting ANXA3 to combat HCC with anti-ANXA3 monoclonal antibody alone or in combination with sorafenib/regorafenib was investigated ex vivo and in vivo. Results ANXA3 conferred HCC cells with resistance to sorafenib. ANXA3 was found enriched in sorafenib-resistant HCC cells and patient-derived xenografts. Mechanistically, overexpression of ANXA3 in sorafenib-resistant HCC cells suppressed PKCδ/p38 associated apoptosis and activated autophagy for cell survival. Clinically, ANXA3 expression correlated positively with the autophagic marker LC3B in HCC and was associated with a worse overall survival in patients who went on to receive sorafenib treatment. Anti-ANXA3 monoclonal antibody therapy combined with sorafenib/regorafenib impaired tumor growth in vivo and significantly increased survival. Conclusion Anti-ANXA3 therapy in combination with sorafenib/regorafenib represents a novel therapeutic strategy for HCC treatment. ANXA3 represents a useful predictive biomarker to stratify patients with HCC for sorafenib treatment. Lay summary This study represents the most extensive pre-clinical characterization of anti-ANXA3 monoclonal antibodies for the treatment of hepatocellular carcinoma to date. These results support the clinical trial development of anti-ANXA3 antibodies in combination with sorafenib/regorafenib. Further studies will optimize patient target selection and identify the best treatment combinations.
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ANXA3/JNK Signaling Promotes Self-Renewal and Tumor Growth, and Its Blockade Provides a Therapeutic Target for Hepatocellular Carcinoma
Stem cell reports, 2015Co-Authors: Man Tong, Steve T. Luk, Terence K. Lee, Tsun Ming Fung, Chi Ho Lin, Judy Wai Ping Yam, Kwok Wah Chan, Bo-jian ZhengAbstract:Frequent tumor relapse in hepatocellular carcinoma (HCC) has been commonly attributed to the presence of residual cancer stem cells (CSCs) after conventional treatments. We have previously identified and characterized CD133 to mark a specific CSC subset in HCC. In the present study, we found endogenous and secretory Annexin A3 (ANXA3) to play pivotal roles in promoting cancer and stem cell-like features in CD133+ liver CSCs through a dysregulated JNK pathway. Blockade of ANXA3 with an anti-ANXA3 monoclonal antibody in vitro as well as in human HCC xenograft models resulted in a significant reduction in tumor growth and self-renewal. Clinically, ANXA3 expression in HCC patient sera closely associated with aggressive clinical features. Our results suggest that ANXA3 can serve as a novel diagnostic biomarker and that the inhibition of ANXA3 may be a viable therapeutic option for the treatment of CD133+ liver-CSC-driven HCC.
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Abstract 221: Annexin A3 is a therapeutic target for CD133+ liver cancer stem cells
Tumor Biology, 2014Co-Authors: Man Tong, Chun Ming Fung, Xin Yuan GuanAbstract:Proceedings: AACR Annual Meeting 2014; April 5-9, 2014; San Diego, CA Frequent tumor relapse in hepatocellular carcinoma (HCC) has been commonly attributed to the failure to completely eradicate cancer stem cells (CSCs) in the tumor residues by conventional treatments. We have previously reported that the tumor growth of HCC is fuelled, at least in part, by a small subset of CSCs marked by the CD133 surface phenotype. Our present study aims 1) to delineate the molecular mechanism by which CD133+ liver CSCs mediate HCC tumor formation and progression; and 2) to develop a novel diagnostic / prognostic biomarker and targeted therapy for HCC detection and treatment. RNA-Seq profiling was employed to compare the gene expression profiles between sorted CD133+ and CD133- subsets isolated from HCC cell lines Huh7 and PLC8024. Annexin A3 (ANXA3) was identified as the most significantly up-regulated secretory protein in the CD133+ subset. Validation in additional HCC cell lines and clinical samples likewise showed ANXA3 to be preferentially expressed in the CD133+ subset. Up-regulation of ANXA3 in both endogenous and secretory forms was tightly associated with advanced tumor stages. In addition, quantification of serum ANXA3 provides a novel biomarker with high specificity and sensitivity for HCC diagnosis. Functional studies involving lentiviral-based knockdown and overexpression approaches found ANXA3 to regulate cancer and stem cell-like properties including tumor initiation, migration, invasion, angiogenesis, self-renewal and resistance to chemotherapy and apoptosis. Subsequent gene expression profiling by cDNA microarray found ANXA3 to be functionally involved in driving CD133+ CSCs via a deregulated JNK/AP-1 pathway. In light of the functional significance and clinical relevance of ANXA3 in HCC, we subsequently developed a novel neutralizing monoclonal antibody specific against ANXA3 and tested for its application as a therapeutic treatment against HCC. In vitro functional studies showed that ANXA3 neutralizing antibody treatment dramatically reduced cell proliferation, sphere formation, migration, invasion, angiogenesis, and sensitized cells to apoptosis and cisplatin treatment in vitro. This observation was further confirmed in vivo where neutralizing antibody treatment attenuated tumor growth, concomitant with a reduced CD133+ subset in the residual xenografts. Mechanistically, treatment of HCC cells with ANXA3 neutralizing antibody in vitro and in vivo, similarly led to a suppressed JNK pathway. Taken together, ANXA3 confers both cancer and stem cell-like properties in CD133+ liver CSCs via JNK pathway. ANXA3 neutralizing antibody possesses therapeutic effect in HCC by eliminating CD133+ liver CSCs. We believe that ANXA3 could serve as a potential therapeutic target to attain complete eradication of HCC. Citation Format: Man Tong, Chun Ming Fung, Xin Yuan Guan, Stephanie Ma. Annexin A3 is a therapeutic target for CD133+ liver cancer stem cells. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 221. doi:10.1158/1538-7445.AM2014-221
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Clinical relevance, functional significance and therapeutic implication of Annexin A3 in CD133⁺ liver cancer stem cells driven hepatocellular carcinoma
1Co-Authors: Man TongAbstract:Hepatocellular carcinoma (HCC) is the third leading cause of mortality in Southeast Asia and in Hong Kong. Despite advances in treatment, the prognosis of advanced HCC patients remains dismal. Contemporary challenge in treating HCC has been the common chemotherapy resistance and recurrence after therapy, all of which have been reported to be associated with stem cell-like behaviors of cancer cells. We and others have previously reported that the tumor growth of HCC is fuelled in part by a subset of cells called cancer stem cells (CSCs) marked by the CD133 phenotype and bearing features that include the ability to self-renew, differentiate, initiate tumors in vivo and resist chemotherapy. Despite our growing knowledge of the importance of 〖CD133〗^+ liver CSCs in hepatocarcinogenesis, there is still a lack of diagnostic markers and therapeutic strategies targeted specifically at this subset of cells. Therefore, the identification of valid targets and the development of new therapies are urgently needed for the more effective clinical management of this deadly disease. By RNA Sequencing profiling of sorted 〖CD133〗^+ liver CSCs and 〖CD133〗^- HCC cells isolated from Huh7 and PLC8024, we identified Annexin A3 (ANXA3) to be the most commonly up-regulated gene which encodes for secretory proteins in the 〖CD133〗^+ subset. This observation was further validated in CD133 sorted cells isolated from additional HCC cell lines and clinical samples. Clinically, up-regulation of both endogenous and secretory ANXA3 was found to be tightly associated with advanced tumor stages. Importantly, the diagnostic value of secretory ANXA3 was found to be more superior to the commonly used biomarker alpha-fetoprotein (AFP). Subsequent functional studies involving lentiviral-based knockdown and overexpression found endogenous ANXA3 to regulate cancer and stem cell-like properties including tumor initiation, migration, invasion, metastasis, angiogenesis, self-renewal and resistance to chemotherapy and apoptosis. Likewise, co-culture of ANXA3-containing conditioned medium or recombinant ANXA3 potentiated cancer and stem cell-like properties in HCC cells. In view of the clinical significance and functional relevance of ANXA3 in HCC, we subsequently developed a novel monoclonal antibody specific against ANXA3 and tested its ability to inhibit oncogenic functions. Results from our in vitro functional studies showed that anti-ANXA3 monoclonal antibody could reduce the ability of HCC cells to confer cancer and stem cell-like features, while administration of the neutralizing antibody in combination with cisplatin in immunodeficient mice bearing HCC xenografts resulted in a significant reduction in tumor growth and self-renewal, concomitant with a decrease in CD133 expression. The mechanistic pathway by which ANXA3 drives HCC was subsequently elucidated. We found internalization of secretory ANXA3 in HCC cells to be dependent on caveolaemediated endocytosis. Gene expression profiling and molecular studies found internalized ANXA3 to drive HCC through a deregulated JNK pathway. This observation was further substantiated by rescue experiments involving a JNK inhibitor. Taken together, ANXA3 exerts oncogenic effects in 〖CD133〗^+ liver CSCs via the JNK pathway and anti-ANXA3 neutralizing antibody holds therapeutic potential for HCC patients. We believe that ANXA3 could serve as a novel diagnostic biomarker and therapeutic target in HCC.published_or_final_versionAnatomyDoctoralDoctor of Philosoph
Wangyuan Zou - One of the best experts on this subject based on the ideXlab platform.
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microglial Annexin A3 downregulation alleviates bone cancer induced pain through inhibiting the hif 1α vascular endothelial growth factor signaling pathway
Pain, 2020Co-Authors: Zengli Zhang, Qulian Guo, Meiling Deng, Jiangju Huang, Manyu Xing, Jian Wang, Wangyuan ZouAbstract:Bone cancer-induced pain (BCP) is a challenging clinical problem because traditional therapies are often only partially effective. Annexin A3 (ANXA3) is highly expressed in microglia in the spinal cord, and its expression is upregulated during BCP. However, the roles of microglial ANXA3 in the development and maintenance of BCP and the underlying molecular mechanisms remain unclear. This study was performed on male mice using a metastatic lung BCP model. Adeno-associated virus shANXA3 (AAV-shANXA3) was injected intrathecally 14 days before and 7 days after bone cancer induction, and relevant pain behaviors were assessed by measuring the paw withdrawal mechanical threshold, paw withdrawal thermal latency, and spontaneous hind limb lifting. ANXA3 protein expression was downregulated in microglial N9 cells by lentiviral transfection (LV-shANXA3). ANXA3, hypoxia-inducible factor-1α (Hif-1α), vascular endothelial growth factor (VEGF) expression levels, and Hif-1α transactivation activity regulated by ANXA3 were measured. As a result, ANXA3 was expressed in microglia, and its expression significantly increased during BCP. ANXA3 knockdown reversed pain behaviors but did not prevent pain development. Moreover, ANXA3 knockdown significantly reduced Hif-1α and VEGF expression levels in vitro and in vivo. And overexpression of Hif-1α or VEGF blocked the effects of AAV-shANXA3 on BCP. ANXA3 knockdown in N9 cells significantly decreased the p-PKC protein expression in the cocultured neurons. Finally, ANXA3 overexpression significantly increased Hif-1α transactivation activity in 293T cells. Therefore, microglial ANXA3 downregulation alleviates BCP by inhibiting the Hif-1α/VEGF signaling pathway, which indicates that ANXA3 may be a potential target for the treatment of BCP.
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Microglial Annexin A3 downregulation alleviates bone cancer-induced pain through inhibiting the Hif-1α/vascular endothelial growth factor signaling pathway.
Pain, 2020Co-Authors: Zengli Zhang, Qulian Guo, Meiling Deng, Jiangju Huang, Manyu Xing, Jian Wang, Wangyuan ZouAbstract:Bone cancer-induced pain (BCP) is a challenging clinical problem because traditional therapies are often only partially effective. Annexin A3 (ANXA3) is highly expressed in microglia in the spinal cord, and its expression is upregulated during BCP. However, the roles of microglial ANXA3 in the development and maintenance of BCP and the underlying molecular mechanisms remain unclear. This study was performed on male mice using a metastatic lung BCP model. Adeno-associated virus shANXA3 (AAV-shANXA3) was injected intrathecally 14 days before and 7 days after bone cancer induction, and relevant pain behaviors were assessed by measuring the paw withdrawal mechanical threshold (PWMT), paw withdrawal thermal latency (PWTL), and spontaneous hindlimb lifting. ANXA3 protein expression was downregulated in microglial N9 cells by lentiviral transfection (LV-shANXA3). ANXA3, hypoxia-inducible factor-1α (Hif-1α), vascular endothelial growth factor (VEGF) expression levels and Hif-1α transactivation activity regulated by ANXA3 were measured. As a result, ANXA3 was expressed in microglia, and its expression significantly increased during BCP. ANXA3 knockdown reversed pain behaviors but didn't prevent pain development. Moreover, ANXA3 knockdown significantly reduced Hif-1α and VEGF expression levels in vitro and in vivo. And overexpression of Hif-1α or VEGF blocked the effects of AAV-shANXA3 on BCP. ANXA3 knockdown in N9 cells significantly decreased the p-PKC protein expression in the co-cultured neurons. Finally, ANXA3 overexpression significantly increased Hif-1α transactivation activity in 293T cells. Therefore, microglial ANXA3 downregulation alleviates BCP by inhibiting the Hif-1α/VEGF signaling pathway, which indicates that ANXA3 may be a potential target for the treatment of BCP.
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Proteomic Identification of an Upregulated Isoform of Annexin A3 in the Spinal Cords of Rats in a Neuropathic Pain Model.
Frontiers in neuroscience, 2017Co-Authors: Wangyuan Zou, Zongbin Song, Tao Zhong, Yingqi Weng, Changsheng Huang, Chuanlei Zhang, Xianquan Zhan, Qulian GuoAbstract:Neuropathic pain (NP) is induced by nerve damage or a disturbance in the peripheral or central nervous systems. Nerve damage causes the activation of sensitizing mechanisms in the peripheral and central nervous systems, which induces transcriptional and post-transcriptional alterations in sensory nerves. However, the underlying mechanisms of NP remain elusive. In the study, Two-dimensional gel electrophoresis (2DGE)-based comparative proteomics identified 38 differential gel spots, and 15 differentially expressed protein (DEPs) between the sham and the chronic constriction injury (CCI) -induced neuropathic pain rats. Of them, Annexin A3 (ANXA3) was significantly increased after CCI with Western blot analysis and immunofluorescence imaging. A lentivirus delivering ANXA3 shRNA (LV-shANXA3) was administered intrathecally to determine the analgesic effects of ANXA3 on allodynia and hyperalgesia in a CCI-induced neuropathic pain model in rats. Further study showed that LV-shANXA3 reversed the upregulation of ANXA3, alleviated CCI-induced mechanical allodynia and thermal hyperalgesia. The study indicated that ANXA3 may play an important role in neuropathic pain.
Lingya Pan - One of the best experts on this subject based on the ideXlab platform.
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Annexin A3 Is a Potential Predictor of Platinum Resistance in Epithelial Ovarian Cancer Patients in a Prospective Cohort
Journal of Cancer, 2015Co-Authors: Ying Jin, Jie Yin, Li-ping Feng, Xiang Jiang, Yongxue Wang, Zi-ping Yang, Lingya PanAbstract:Epithelial ovarian cancer (EOC) is the leading cause of death among gynecological malignancies and is rarely cured in the recurrent setting, mainly because of progressive chemoresistance, especially platinum resistance. In our previous studies, the platinum-resistance-related protein, Annexin A3, was selected by comparative proteomics. In this study, we detected serum Annexin A3 levels using a self-developed chemiluminescence immunoassay kit in a prospective EOC patient cohort. We also evaluated the capacity of serum Annexin A3 levels to predict platinum resistance. Serum Annexin A3 levels in healthy women exhibited a similar normal distribution (Z=0.723, P=0.673), allowing determination of a normal cutoff level of 0.11-1.45 ng/mL. Of the 89 EOC patients, 21 were platinum resistant and 68 were platinum sensitive. Residual disease after primary surgery (p=0.004) and serum Annexin A3 levels (p=0.036) were both independent factors associated with platinum resistance. The AUC was 0.733 (95% confidence interval (CI), 0.627-0.823). The optimal cutoff value for serum Annexin A3 levels was 2.05 ng/mL. Multivariate logistic analysis showed that expression of Annexin A3 as assessed by immunohistochemistry (P=0.005) and residual tumor size (P=0.000) had a significant influence on platinum resistance. The AUC of ROC curve of Annexin A3 expression by immunohistochemistry was 0.664 (95% CI, 0.554-0.763) and the cut off value was “>=moderate scores”. In conclusion, we demonstrate that Annexin A3 is a secreted protein that may be measured in the peripheral blood using a self-developed, chemiluminescence immunoassay kit. Serum Annexin A3 levels may be a potential predictor of platinum resistance in epithelial ovarian cancer patients.
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Secretion of Annexin A3 from ovarian cancer cells and its association with platinum resistance in ovarian cancer patients
Journal of cellular and molecular medicine, 2012Co-Authors: Jie Yin, Xuedong Yan, Ning Mao, Yili Yang, Xin Yao, Yongli Zhang, Ying Shan, Lingya PanAbstract:Early detection of resistance to platinum-based therapy is critical for improving the treatment of ovarian cancers. We have previously found that increased expression of Annexin A3 is a mechanism for platinum resistance in ovarian cancer cells. Here we demonstrate that Annexin A3 can be detected in the culture medium of ovarian cancer cells, particularly these cells that express high levels of Annexin A3. Levels of Annexin A3 were then determined in sera from ovarian cancer patients using an enzyme-linked immunosorbent assay. Compared with those from normal donors, sera from ovarian cancer patients contain significantly higher levels of Annexin A3. Furthermore, serum levels of Annexin A3 were significantly higher in platinum-resistant patients than in platinum-sensitive patients. To gain insight into the mechanism of secretion, the ovarian cancer cell lines were examined using both transmission electron microscopy and immunoelectron microscopy. Compared with parent cells, there are significantly more vesicles in the cytoplasm of ovarian cancer cells that express high levels of Annexin A3, and at least some vesicles are Annexin A3-positive. Moreover, some vesicles appear to be fused with the cell membrane, suggesting that Annexin A3 secretion may be associated with exocytosis and the release of exosomes. This is supported by our observation that ovarian cancer cells expressing higher levels of Annexin A3 released increased numbers of exosomes. Furthermore, Annexin A3 can be detected in exosomes released from cisplatin-resistant cells (SKOV3/Cis) by immunoblotting and immunoelectron microscopy.
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Increased Expression of Annexin A3 Is a Mechanism of Platinum Resistance in Ovarian Cancer
Cancer research, 2010Co-Authors: Xuedong Yan, Jie Yin, Huiyu Yao, Ning Mao, Yili Yang, Lingya PanAbstract:Resistance to platinum drugs has emerged as a major obstacle in the treatment of ovarian cancers. Through proteomic analysis, we have found that the expression of Annexin A3, a member of the Ca2+ and phospholipid-binding Annexin family, is significantly increased in platinum-resistant ovarian cell lines. Anti–Annexin A3 immunostaining indicated that cancers from platinum-resistant patients also possess higher levels of Annexin A3 than those from platinum-sensitive patients. Although expression of Annexin A3 made susceptible ovarian cancer cells more resistant to platinum, expression of antisense Annexin A3 downregulated its expression and rendered the resistant cells more sensitive to platinum. In athymic mice, the growth of tumors from inoculated SKOV3 cells was inhibited by the administration of platinum, whereas tumors from Annexin A3–expressing SKOV3/Ann were resistant to platinum treatment. Interestingly, the intracellular platinum concentration and platinum-DNA binding are significantly lower in Annexin A3–overexpressing cells than those in parental cells. The lower cisplatin concentration was also accompanied by reduced induction of p53, which could be restored by downregulation of Annexin A3. These results indicate that increased expression of Annexin A3 is a mechanism of platinum resistance in ovarian cancer. It seems to act by preventing uptake or accumulation of platinum in cells. Therefore, it is conceivable that Annexin A3 could be a target for therapeutic intervention and may also serve as a biomarker for drug resistance in ovarian cancer patients. Cancer Res; 70(4); 1616–24
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Identification of platinum-resistance associated proteins through proteomic analysis of human ovarian cancer cells and their platinum-resistant sublines.
Journal of proteome research, 2007Co-Authors: Xuedong Yan, Lingya Pan, Ye Yuan, Jing-he Lang, Ning MaoAbstract:Chemoresistance is a major therapeutic obstacle in cancer patients, and the mechanisms of drug resistance are not fully understood. In the present study, we established platinum-resistant human ovarian cancer cell lines and identified differentially expressed proteins related to platinum resistance. The total proteins of two sensitive (SKOV3 and A2780) and four resistant (SKOV3/CDDP, SKOV3/CBP, A2780/CDDP, and A2780/CBP) human ovarian cancer cell lines were isolated by two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were identified using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). In total, 57 differential protein spots were identified, and five proteins, including Annexin A3, destrin, cofilin 1, Glutathione-S-transferase omega 1 (GSTO1-1), and cytosolic NADP+-dependent isocitrate dehydrogenase (IDHc), were found to be co-instantaneous significance compared with their parental cells. The expression of the five proteins was validated by quantitative PCR and western blot, and the western blot results showed complete consistency with proteomic techniques. The five proteins are hopeful to become candidates for platinum resistance. These may be useful for further study of resistance mechanisms and screening of resistant biomarkers.
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Proteomic analysis of human ovarian cancer cell lines and their platinum-resistant clones
Zhonghua fu chan ke za zhi, 2006Co-Authors: Xuedong Yan, Lingya PanAbstract:OBJECTIVE To perform comparative proteomic analysis of human ovarian cancer cell lines for detecting platinum-resistance associated proteins. METHODS The total proteins of two sensitive (SKOV3 and A2780) and four resistant (SKOV3/CDDP, SKOV3/CBP, A2780/CDDP and A2780/CBP) human ovarian cancer cell lines were separated by two-dimensional gel electrophoresis (2-DE). The differentially expressed proteins were analyzed using image analysis software, stained with Coomassie Brilliant Blue, then identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and database searching. The mRNA and protein levels of the differentially expressed protein which was most significant in all of the four resistant cell lines were validated by RT-PCR and western blotting, respectively. RESULTS Five proteins were found to be significant in four cell lines. Annexin A3 and destrin were up-regulated and nicotinamide-adenine dinucleotide phosphate (NADP)-dependent isocitrate dehydrogenase 1 was down-regulated in all the four resistant samples. Glutathione transferase omega 1 had an increased expression in the other three resistant cell lines except for SKOV3/CBP in which its expression was not changed. However, cofilin 1 represented a different trend. In the two resistant sublines of SKOV3, cofilin 1 had a down-regulation, but it had an up-regulation in the cell lines induced from SKOV3. The expression of Annexin A3 was up-regulated by 3 - 20 fold and the results of RT-PCR and western blotting showed complete consistency with that by 2-DE. CONCLUSIONS Proteomic techniques are useful to the identification of the resistance-associated proteins in ovarian cancer platinum-resistant cell lines and five candidates have been found. The five differential proteins might become hopeful candidate biomarkers for resistance.
Hiroshi Nishiura - One of the best experts on this subject based on the ideXlab platform.
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Participation of delta Annexin A3 in the ribosomal protein S19 C‐terminus‐dependent inhibitory mechanism of the neutrophil C5a receptor through delta lactoferrin
Pathology International, 2017Co-Authors: Koji Yamanegi, Naoko Yamada, Keiji Nakasho, Hiroshi NishiuraAbstract:Although C5a receptor (C5aR) interacting with its agonist C5a promotes acute inflammation during the initiation phase, the roles of the recycling C5aR during the resolution phase are still unclear. We found that C5aR interacted with its antagonist/agonist ribosomal protein S19 (RP S19) polymer or a RP S19 polymer functional analogue S-tagged C5a/RP S19, which connects an RP S19 C-terminus (IAGQVAAANKKH) to the S-tagged C5a C-terminus, promoted acute inflammation at the resolution phase via an activation of the apoptosis-inducing transcription factor delta lactoferrin (δLf) in neutrophils and the membrane mobilizing factor full-length Annexin A3 (ANXA3) in macrophages. To confirm the antagonistic system of the recycling C5aR, S-tagged δLf-coupled BrCN-activated Sepharose 4B beads were incubated with cytoplasmic proteins and identified a neutrophil-specific δANXA3 via pull-down experiments. The S-tagged C5a/RP S19-induced agonistic functions in macrophage-like cells that were differentiated from human promyelocytic leukemia HL-60 cells by phorbol-12-myristate-13-acetate were suppressed by δLf and δANXA3 co-overexpression. δANXA3 seems to participate in the antagonistic system of the neutrophil C5aR involving IAGQVAAANKKH and δLf. Most likely, δANXA3 works as antagonist for the recycling C5aR on neutrophils during the resolution phase of acute inflammation.
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Participation of delta Annexin A3 in the ribosomal protein S19 C-terminus-dependent inhibitory mechanism of the neutrophil C5a receptor through delta lactoferrin.
Pathology international, 2017Co-Authors: Koji Yamanegi, Naoko Yamada, Keiji Nakasho, Hiroshi NishiuraAbstract:Although C5a receptor (C5aR) interacting with its agonist C5a promotes acute inflammation during the initiation phase, the roles of the recycling C5aR during the resolution phase are still unclear. We found that C5aR interacted with its antagonist/agonist ribosomal protein S19 (RP S19) polymer or a RP S19 polymer functional analogue S-tagged C5a/RP S19, which connects an RP S19 C-terminus (IAGQVAAANKKH) to the S-tagged C5a C-terminus, promoted acute inflammation at the resolution phase via an activation of the apoptosis-inducing transcription factor delta lactoferrin (δLf) in neutrophils and the membrane mobilizing factor full-length Annexin A3 (ANXA3) in macrophages. To confirm the antagonistic system of the recycling C5aR, S-tagged δLf-coupled BrCN-activated Sepharose 4B beads were incubated with cytoplasmic proteins and identified a neutrophil-specific δANXA3 via pull-down experiments. The S-tagged C5a/RP S19-induced agonistic functions in macrophage-like cells that were differentiated from human promyelocytic leukemia HL-60 cells by phorbol-12-myristate-13-acetate were suppressed by δLf and δANXA3 co-overexpression. δANXA3 seems to participate in the antagonistic system of the neutrophil C5aR involving IAGQVAAANKKH and δLf. Most likely, δANXA3 works as antagonist for the recycling C5aR on neutrophils during the resolution phase of acute inflammation.
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Annexin A3 plays a role in cytoplasmic calcium oscillation by extracellular calcium in the human promyelocytic leukemia HL-60 cells differentiated by phorbol-12-myristate-13-acetate
Experimental and Molecular Pathology, 2014Co-Authors: Hiroshi Nishiura, Koji Yamanegi, Naoko Yamada, Mutsuki Kawabe, Nahoko Kato-kogoe, Keiji NakashoAbstract:Abstract The roles of Annexin A3 (ANXA3) in macrophages are not fully understood. In contrast to C5a, we have demonstrated that C-terminal ribosomal protein S19 (RP S19)-tagged S-tagged C5a (S-tagged C5a/RP S19) raises an alternative cytoplasmic calcium oscillation by extracellular calcium during macrophage migration into apoptotic cells. We here differentiated human promyelocytic leukemia HL-60 cells bearing with either control sense RNA and shRNA for ANXA3 mRNA or a vector cDNA with or without ANXA3 cDNA into macrophage-like cells by phorbol-12-myristate-13-acetate and found that a fluorescence ratio (340 nm/380 nm) upon the S-tagged C5a/RP S19-induced alternative cytoplasmic calcium oscillation by extracellular calcium was an equilateral association with a dose of ANXA3. Moreover, the ANXA3-dependent modification was partially reflected upon the S-tagged C5a-induced classical cytoplasmic calcium oscillation by both intracellular calcium and extracellular calcium. ANXA3 seems to extend the C5aR-mediated cytoplasmic calcium oscillation by extracellular calcium at least in the HL-60 macrophage-like cells.
Keiji Nakasho - One of the best experts on this subject based on the ideXlab platform.
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Participation of delta Annexin A3 in the ribosomal protein S19 C‐terminus‐dependent inhibitory mechanism of the neutrophil C5a receptor through delta lactoferrin
Pathology International, 2017Co-Authors: Koji Yamanegi, Naoko Yamada, Keiji Nakasho, Hiroshi NishiuraAbstract:Although C5a receptor (C5aR) interacting with its agonist C5a promotes acute inflammation during the initiation phase, the roles of the recycling C5aR during the resolution phase are still unclear. We found that C5aR interacted with its antagonist/agonist ribosomal protein S19 (RP S19) polymer or a RP S19 polymer functional analogue S-tagged C5a/RP S19, which connects an RP S19 C-terminus (IAGQVAAANKKH) to the S-tagged C5a C-terminus, promoted acute inflammation at the resolution phase via an activation of the apoptosis-inducing transcription factor delta lactoferrin (δLf) in neutrophils and the membrane mobilizing factor full-length Annexin A3 (ANXA3) in macrophages. To confirm the antagonistic system of the recycling C5aR, S-tagged δLf-coupled BrCN-activated Sepharose 4B beads were incubated with cytoplasmic proteins and identified a neutrophil-specific δANXA3 via pull-down experiments. The S-tagged C5a/RP S19-induced agonistic functions in macrophage-like cells that were differentiated from human promyelocytic leukemia HL-60 cells by phorbol-12-myristate-13-acetate were suppressed by δLf and δANXA3 co-overexpression. δANXA3 seems to participate in the antagonistic system of the neutrophil C5aR involving IAGQVAAANKKH and δLf. Most likely, δANXA3 works as antagonist for the recycling C5aR on neutrophils during the resolution phase of acute inflammation.
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Participation of delta Annexin A3 in the ribosomal protein S19 C-terminus-dependent inhibitory mechanism of the neutrophil C5a receptor through delta lactoferrin.
Pathology international, 2017Co-Authors: Koji Yamanegi, Naoko Yamada, Keiji Nakasho, Hiroshi NishiuraAbstract:Although C5a receptor (C5aR) interacting with its agonist C5a promotes acute inflammation during the initiation phase, the roles of the recycling C5aR during the resolution phase are still unclear. We found that C5aR interacted with its antagonist/agonist ribosomal protein S19 (RP S19) polymer or a RP S19 polymer functional analogue S-tagged C5a/RP S19, which connects an RP S19 C-terminus (IAGQVAAANKKH) to the S-tagged C5a C-terminus, promoted acute inflammation at the resolution phase via an activation of the apoptosis-inducing transcription factor delta lactoferrin (δLf) in neutrophils and the membrane mobilizing factor full-length Annexin A3 (ANXA3) in macrophages. To confirm the antagonistic system of the recycling C5aR, S-tagged δLf-coupled BrCN-activated Sepharose 4B beads were incubated with cytoplasmic proteins and identified a neutrophil-specific δANXA3 via pull-down experiments. The S-tagged C5a/RP S19-induced agonistic functions in macrophage-like cells that were differentiated from human promyelocytic leukemia HL-60 cells by phorbol-12-myristate-13-acetate were suppressed by δLf and δANXA3 co-overexpression. δANXA3 seems to participate in the antagonistic system of the neutrophil C5aR involving IAGQVAAANKKH and δLf. Most likely, δANXA3 works as antagonist for the recycling C5aR on neutrophils during the resolution phase of acute inflammation.
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Annexin A3 plays a role in cytoplasmic calcium oscillation by extracellular calcium in the human promyelocytic leukemia HL-60 cells differentiated by phorbol-12-myristate-13-acetate
Experimental and Molecular Pathology, 2014Co-Authors: Hiroshi Nishiura, Koji Yamanegi, Naoko Yamada, Mutsuki Kawabe, Nahoko Kato-kogoe, Keiji NakashoAbstract:Abstract The roles of Annexin A3 (ANXA3) in macrophages are not fully understood. In contrast to C5a, we have demonstrated that C-terminal ribosomal protein S19 (RP S19)-tagged S-tagged C5a (S-tagged C5a/RP S19) raises an alternative cytoplasmic calcium oscillation by extracellular calcium during macrophage migration into apoptotic cells. We here differentiated human promyelocytic leukemia HL-60 cells bearing with either control sense RNA and shRNA for ANXA3 mRNA or a vector cDNA with or without ANXA3 cDNA into macrophage-like cells by phorbol-12-myristate-13-acetate and found that a fluorescence ratio (340 nm/380 nm) upon the S-tagged C5a/RP S19-induced alternative cytoplasmic calcium oscillation by extracellular calcium was an equilateral association with a dose of ANXA3. Moreover, the ANXA3-dependent modification was partially reflected upon the S-tagged C5a-induced classical cytoplasmic calcium oscillation by both intracellular calcium and extracellular calcium. ANXA3 seems to extend the C5aR-mediated cytoplasmic calcium oscillation by extracellular calcium at least in the HL-60 macrophage-like cells.