• Title/Summary/Keyword: XAF1

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XIAP Associated Factor 1 (XAF1) Represses Expression of X-linked Inhibitor of Apoptosis Protein (XIAP) and Regulates Invasion, Cell Cycle, Apoptosis, and Cisplatin Sensitivity of Ovarian Carcinoma Cells

  • Zhao, Wen-Jing;Deng, Bo-Ya;Wang, Xue-Mei;Miao, Yuan;Wang, Jian-Nan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.6
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    • pp.2453-2458
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    • 2015
  • Background: X-linked inhibitor of apoptosis protein (XIAP) associated factor 1 (XAF1) exhibits aberrantly low or absent expression in various human malignancies, closely associated with anti-apoptosis and overgrowth of cancer cells. However, limited attention has been directed towards the contribution of XAF1 to invasion, apoptosis, and cisplatin (DDP)-resistance of epithelial ovarian cancer (EOC) cells. This study aimed to evaluate the potential effects of XAF1 on invasion, cell cycle, apoptosis, and cisplatin-resistance by overexpressing XAF1 in SKOV-3 and SKOV-3/DDP cells. Methods and Results: The pEGFP-C1-XAF1 plasmid was transfected into SKOV-3 and SKOV-3/DDP cells, and the expression of XAF1 at both mRNA and protein levels was analyzed by reverse transcription-PCR and Western blotting. Overexpression of XAF1 suppressed XIAP expression in both SKOV-3 and SKOV-3/DDP cells. Transwell invasion assays demonstrated that XAF1 exerted a strong anti-invasive effect in XAF1-overexpressing cells. Moreover, flow cytometry analysis revealed that XAF1 overexpression arrested the cell cycle at G0/G1 phase, and cell apoptosis analysis showed that overexpression of XAF1 enhanced apoptosis of SKOV-3 and SKOV-3/DDP cells apparently by activating caspase-9 and caspase-3. Furthermore, MTT assay confirmed a dose-dependent inhibitory effect of cisplatin in the tested tumor cells, and overexpression of XAF1 increased the sensitivity of SKOV-3 and SKOV-3/DDP cells to cisplatin-mediated antiproliferative effects. Conclusions: In summary, our data indicated that overexpression of XAF1 could suppress XIAP expression, inhibit invasion, arrest cell cycle, promote apoptosis, and confer cisplatin-sensitivity in SKOV-3 and SKOV-3/DDP cells. Therefore, XAF1 may be further assessed as a potential target for the treatment of both cisplatin-resistant and non-resistant EOCs.

Experimental Study on Inhibition Effects of the XAF1 Gene against Lung Cancer Cell Proliferation

  • Yang, Wen-Tao;Chen, Dong-Lai;Zhang, Fu-Quan;Xia, Ying-Chen;Zhu, Rong-Ying;Zhou, Duan-Shan;Chen, Yong-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.18
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    • pp.7825-7829
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    • 2014
  • Objective: To investigate the effect of high expression of XAF1 in vivo or in vitro on lung cancer cell growth and apoptosis. Methods: 1. The A549 human lung cancer cell line was transfected with Ad5/F35 - XAF1, or Ad5/F35 - Null at the same multiplicity of infection (MOI); (hereinafter referred to as transient transfected cell strain); XAF1 gene mRNA and protein expression was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting respectively. 2. Methyl thiazolyl tetrazolium (MTT) and annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis before and after infection of Ad5/F35 - XAF1 with Western blotting for apoptosis related proteins, caspase 3, caspase - 8 and PARP. 3. After the XAF1 gene was transfected into lung cancer A549 cells by lentiviral vectors, and selected by screening with Blasticidin, reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were applied to detect mRNA and protein expression, to establish a line with a stable high expression of XAF1 (hereinafter referred to as stable expression cell strain). Twenty nude mice were randomly divided into groups A and B, 10 in each group: A549/XAF1 stable expression cell strain was subcutaneously injected in group A, and A549/Ctrl stable cell line stable expression cell strain in group B (control group), to observe transplanted tumor growth in nude mice. Results: The mRNA and protein expression of XAF1 in A549 cells transfected by Ad5/F35 - XAF1 was significantly higher than in the control group. XAF1 mediated by adenovirus vector demonstrated a dose dependent inhibition of lung cancer cell proliferation and induction of apoptosis. This was accompanied by cleavage of caspase -3, -8, -9 and PARP, suggesting activation of intrinsic or extrinsic apoptotic pathways. A cell strain of lung cancer highly expressing XAF1 was established, and this demonstrated delayed tumor growth after transplantation in vivo. Conclusion: Adenovirus mediated XAF1 gene expression could inhibit proliferation and induce apoptosis in lung cancer cells in vitro; highly stable expression of XAF1 could also significantly inhibit the growth of transplanted tumors in nude mouse, with no obvious adverse reactions observed. Therefore, the XAF1 gene could become a new target for lung cancer treatment.

Characterization of immune gene expression in rock bream (Oplegnathus fasciatus) kidney infected with rock bream iridovirus (RBIV) using microarray

  • Myung-Hwa Jung;Sung-Ju Jung
    • Journal of fish pathology
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    • v.36 no.2
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    • pp.191-211
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    • 2023
  • Rock bream iridovirus (RBIV) causes high mortality and economic losses in rock bream (Oplegnathus fasciatus) aquaculture industry in Korea. Although, the immune responses of rock bream under RBIV infection have been studied, there is not much information at the different stages of infection (initial, middle and recovery). Gene expression profiling of rock bream under different RBIV infection stages was investigated using a microarray approaches. In total, 5699 and 6557 genes were significantly up- or down-regulated over 2-fold, respectively, upon RBIV infection. These genes were grouped into categories such as innate immune responses, adaptive immune responses, complements, lectin, antibacterial molecule, stress responses, DNA/RNA binding, energy metabolism, transport and cell cycle. Interestingly, hemoglobins (α and β) appears to be important during pathogenesis; it is highly up-regulated at the initial stage and is gradually decreased when the pathogen most likely multiplying and fish begin to die at the middle or later stage. Expression levels were re-elevated at the recovery stage of infection. Among up-regulated genes, interferon-related genes were found to be responsive in most stages of RBIV infection. Moreover, X-linked inhibitor of apoptosis (XIAP)-associated factor 1 (XAF1) expression was high, whereas expression of apoptosis-relate genes were low. In addition, stress responses were highly induced in the virus infection. The cDNA microarray data were validated using quantative real-time PCR. Our results provide novel inslights into the broad immune responses triggered by RBIV at different infection stages.

Application of X-ray Absorption Spectroscopy (XAS) in the Field of Stabilization of As and Heavy Metal Contaminated Soil (비소 및 중금속 오염토양 안정화 분야에서의 X선 흡수분광법(XAS) 활용)

  • Lim, Jung Eun;Moon, Deok Hyun;Kim, Kwon-Rae;Ok, Yong Sik
    • Journal of Applied Biological Chemistry
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    • v.58 no.1
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    • pp.65-74
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    • 2015
  • X-ray absorption fine structure (XAFS) analysis using X-ray absorption spectroscopy is being applied as a state-of-the-art method in a wide range of disciplines. This review article summarizes the overall procedure of XAFS analysis from the preparation of soil samples to the analysis of data in X-ray absorption near edge structure (XANES) region and extended Xray absorption fine structure (EXAFS) region. The previous studies on application of XANES and EXAFS techniques in environmental soil science field are discussed and classified them according to metal(loid)s (As, Cd, Cu, Ni, Pb, and Zn). A significant number of previous studies of XAFS application in the environmental soil science field have focused on the identification of Pb chemical species in soil. Moreover, XANES and EXAFS techniques have been widely used to investigate the contamination source via identification of metal species. Similarly, these techniques were applied to identify the mechanisms of metal stabilization in soil after application of various amendments, phytoremediation, etc.