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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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Objective To investigate the anticancer activity of dendritic cells(DC)transfected by recombinant adeno-associated virus(rAAV)-mediated carcinoembryonic antigen(CEA)gene in colorectal cancer treatment.Methods The recombinant plasmid rAAV/CEA was constructed and the rAAV/CEA virus stock was prepared.DC precursors were isolated from peripheral blood mononuclear cells(PBMC)by density gradient centrifugation and transfected with rAAV/CEA virus stock.The CEA expression,chromosome integration and efficiency of rAAV/CEA virus were determined by PCR,Southern blot and flow cytometery.DC precursor cells were induced by granulocyte macrophage colony-stimulating factor(GM-CSF),interleukin-4(IL-4)and tumor necrosis factor-?(TNF-?)to mature DC.The mature DC and T cells were mixed,then cultured with interleukin-2(IL-2)and interleukin-7(IL-7)resulting in generation of cytotoxic T lymphocyte(CTL),whose lethal effects on cancer cells were determined by 51chromium(51Cr)release assay.3H incorporation method was employed to detect the influence of rAAV/CEA transfected DC in the proliferation of activated T cells.Results rAAV medicated CEA gene was transferred into DC precursor successfully and the expression was high.Over 90% DC precursors expressed CEA protein.The viral gene was integrated into the DC chromosome.rAAV/CEA transfected DC could induce specific CTL which could specifically recognize and kill CEA-positive colorectal cancer LoVo cell line,and showed restrictive activity of anti-MHC class I antigen,while it was not able to kill the CEA-negative cancer cells.Conclusion The present study confirms that rAAV/CEA transfected DC vaccine can induce anti-tumor activity in vitro.It paves the way for further study on CEA as a target antigen for gene therapy on colorectal cancer.
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Objective To investigate the best fermentation and induction models of the engineered Escherichia coli, in order to obtain the highest expression level of humanized anti HBsAg Fab. Methods The optimal condition governing the growth of the Escherichia coli, with the flask shaking method and the best fermentation and induction conditions to yield the highest production level were explored, and then E coli were grown in fermentor using the fed batch method following the same principle under flask shaking condition to ensure the best production. Results The data obtained from flask shaking conditions showed that when the induction procedure started the amount of anti HBsAg Fab would reach the highest level at mid log growth phase under the induction condition of 25℃ and 0 2% arabinose. Using the DO stat fed batch method, the OD 600 value of the culture would reach 55 2, which corresponded to 110g/L bacterial wet weight. The biological activity of Fab was proved to have well preserved. Conclusion We established the optimal production technic of HBsAg Fab, and lay a foundation to produce HBsAg Fab on a large scale
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Objective To assess functional affinity of humanized anti-HBsAg Fab in order to provide theoretical basis for the radioimmunotherapy of liver cancer bearing HBV. Methods With cognizance of the advantages of solid phase method in time consumption and convenience, we assessed the functional affinity of engineered anti-HBsAg Fab with non-competitive ELISA method. After a series of trials affirming the best concentration and the best time of HBsAg to cover the plate and the proper binding time of Fab to HBsAg to reach an equilibrium, we plotted the OD standard curve of the binding reaction of Fab and HBsAg using four grades of concentration of HBsAg and a series of consistency of Fab. We got the consistency of Fab at OD50% through the standard curve, and then calculated the affinity by Law of Mass Action. Results The affinity of anti-HBsAg Fab is between 10 7 -10 8 M -1 , which was only smaller by 10 times than that of anti-HBsAg IgG. Conclusion The Fab which we constructed in our lab, can strongly bind to the target antigen, thus providing a theoretical basis for its clinical use.
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Objective Epidermal growth factor receptor (EGFR) plays an important role in the pathogenesis of colorectal cancer (CRC). There are several potential therapies to target EGFR. The detection of EGFR expression is usually performed by immunohistochemistry (IHC) only in the primary tumor. There are a few data regarding the EGFR status in the corresponding distant metastases. The aim of the present study was to correlate EGFR expression in primary tumors and related metastases in order to find out whether EGFR status is different between primary tumors and related metastases. Methods 37 cases (25 male, 12 female) of metastatic colorectal cancer were collected. The site of primary tumor was on colon in 16 patients (43.2%) and on rectum in 21 patients (56.8%). Metastatic sites analyzed were liver in 32 patients (86.5%), inguinal lymph nodes in 5 patients (13.5%). The EGFR status of primary tumors and related metastatic sites were evaluated retrospectively. The expression of EGFR was analyzed by immunohistochemical staining (IHC), EGFR status was defined as positive if the percentage of stained malignant cells was ≥5%, and the expression feature was analyzed. Results EGFR status was positive in 25 primary tumors (67.6%), in which the corresponding metastatic site was negative in 7 cases (28.0%), whereas it was positive in 3 metastases (25.0%) from 12 cases of EGFR-negative primary cancers. The difference between the primary tumors and metastatic lesions had no statistic significance (P=0.23). Conclusion When analyzed by IHC, The difference of EGFR status has no statistic significance between the paired primary tumors and distant metastatic lesions, and 73% of samples show concordant EGFR status.
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Objective To explore the expression of Ezrin in hepatocellular carcinoma(HCC),and the relation of Ezrin expression to E-cadherin,proliferating cell nuclear antigen(PCNA) and clinical features of HCC.Methods Forty-nine cases of HCC were divided into high and low invasive groups according to tumor size,disseminated lesions,distant metastasis,tumor capsule and portal vein tumor thrombus.The expression of Ezrin,E-cadherin and PCNA in normal liver tissues,HCC and adjacent tissues were determined by immunohistochemistry,and the relation of the expression to clinical features was analyzed.Results The Ezrin expression was found in normal liver tissues,HCC and adjacent tissues,and was significantly higher in HCC tissues than in adjacent and normal liver tissues(P