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Short hairpin RNA-mediated knockdown of nuclear factor erythroid 2-like 3 exhibits tumor-suppressing effects in hepatocellular carcinoma cells

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World Journal of Gastroenterology 2019年第10期25卷 1210-1222页

作者： Miao-Mei Yu Yue-Hua Feng Lu Zheng Jun Zhang Guang-Hua Luo Comprehensive Laboratory the Third Affiliated Hospital of Soochow University

BACKGROUND Hepatocellular carcinoma(HCC) is one of the most common malignant tumors with high mortality-to-incidence ratios. Nuclear factor erythroid 2-like 3(NFE2 L3), also known as NRF3, is a member of the cap ‘n' collar basic-region leucine zipper family of transcription factors. NFE2 L3 is involved in the regulation of various biological processes, whereas its role in HCC has not been elucidated.AIM To explore the expression and biological function of NFE2 L3 in HCC.METHODS We analyzed the expression of NFE2 L3 in HCC tissues and its correlation with clinicopathological parameters based on The Cancer Genome Atlas(TCGA) data portal. Short hairpin RNA(shRNA) interference technology was utilized to knock down NFE2 L3 in vitro. Cell apoptosis, clone formation, proliferation, migration,and invasion assays were used to identify the biological effects of NFE2 L3 in BEL-7404 and SMMC-7721 cells. The expression of epithelial-mesenchymal transition(EMT) markers was examined by Western blot analysis.RESULTS TCGA analysis showed that NFE2 L3 expression was significantly positively correlated with tumor grade, T stage, and pathologic stage. The qPCR and Western blot results showed that both the mRNA and protein levels of NFE2 L3 were significantly decreased after shRNA-mediated knockdown in BEL-7404 and SMMC-7721 cells. The shRNA-mediated knockdown of NFE2 L3 could induce apoptosis and inhibit the clone formation and cell proliferation of SMMC-7721 and BEL-7404 cells. NFE2 L3 knockdown also significantly suppressed the migration, invasion, and EMT of the two cell lines.CONCLUSION Our study showed that shRNA-mediated knockdown of NFE2 L3 exhibited tumor-suppressing effects in HCC cells.

关键词： Nuclear factor erythroid 2-like 3 Hepatocellular carcinoma The Cancer Genome Atlas Short hairpin RNA Epithelial-mesenchymal transition

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Bioinformatics and Expression Pattern Analysis of Tomato ns LTP 2-like cDNA full-length Gene Clone

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Journal of Northeast Agricultural University(English Edition) 2019年第1期26卷 28-36页

作者： Zhang Jia He Shan-shan Zhao Ting-ting Jiang Jing-bin Li Jing-fu Xu Xiang-yang College of Horticulture and Landscape Architecture Northeast Agricultural University

TDF1(transcription-drived fragment) was homologous to the predicted S. lycopersicum nonspecific lipid-transfer protein,nsLTP 2-like(91%), and it was significantly upregulated in response to C. fulvum(cladosporium fulvum) infection in tomato plants.In this experiment, the full-length cDNA of nsLTP 2-like was cloned using RACE technology based on the sequence of TDF1(GenBank: JZ717725). A full-length, 625 bp(GenBank: KU366289), cDNA sequence, which with 98% similarity to nsLTP 2-like gene(GenBank: XM015233692) was obtained. This cDNA contains an ORF(open reading frame) with full-length of 345 bp, coding of 114 amino acids, including 12.3% Ala and Gly. Protein molecular weight was 11.51 ku, the isoelectric point(pI) was 8.99, and average overall hydrophilicity was 0.412, with one phosphorylation sites, belonging to volatile acidic nuclear protein. Secondary structure prediction showed that α-Helix accounts for 30.7%, extension chain for 12.28%, β-corner for 9.65%, and random coil for 47.37%. Through comparative analysis of the homology among species, it was found that the amino acid sequence of tomato nsLTP 2-like protein had a high similarity with other plants, and with a specific conserved sequence which might related features in nsLTP 2-like protein. It also be analyzed the gene expression pattern of tomato in different parts and under different stress conditions.The results showed that nsLTP 2-like gene was up-regulated in varying degrees, under the condition of cold stress, exogenous hormone spraying and cladosporium fulvum infection. Therefore, it was speculated that the gene played a role in response to abiotic and biotic stress in tomato.

关键词： tomato nsLTP 2-like cDNA clone bioinformatics analysis expression pattern

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Upregulation of UBAP2L in Bone Marrow Mesenchymal Stem Cells Promotes Functional Recovery in Rats with Spinal Cord Injury

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Current Medical Science 2018年第6期38卷 1081-1089页

作者： Guan-lin LIN Huan WANG Jun DAI Xiao LI Ming GUAN Qing DING Huai-xi WANG Huang FANG Department of Orthopaedics Tongji HospitalTongji Medical CollegeHuazhong University of Science and TechnologyWuhan 430030China

Post-translational modifications of cellular proteins with ubiquitin or ubiquitin-like proteins regulate many cellular processes,such as cell proliferation,differentiation,apoptosis, signal transduction,intercellular immune recognition,inflammatory response,stress response,and DNA repair.Nice4/UBAP2L is an important member in the family of ubiquitin-like proteins,and its biological function remains unknown.This study aimed to investigate the effect of UBAP2L on spinal cord injury (SCI).At first,rat bone marrow mesenchymal stem cells (BMSCs)were infected with adeno-associated virus to induce over-expression of Nice4.Subsequently,the infected BMSCs were transplanted into rats suffering from semi-sectioned SCI.The results showed that the over-expression of Nice4 significantly promoted the proliferation and differentiation of BMSCs. In addition,the transplantation of infected BMSCs into the injured area of SCI rats improved the function repair of SCI.Importantly,the immunohistochemical and hematoxylin-eosin staining and RT-PCR results showed that the number of neuronal cells,oligodendrocytes,and astrocytes was significantly increased in the injured area,along with significantly upregulated expression ofcyclin D1 and p38 mitogen-activated protein kinase (MAPK).Meanwhile,the expression of caspase 3 protein was significantly down-regulated.In conclusion,the over-expression of Nice4 gene can promote the functional recovery in SCI rats by promoting cell proliferation and inhibiting apoptosis. The results of this study indicate an alternative option for the clinical treatment of SCI.

关键词： bone marrow mesenchymal stem cells spinal cord injury ubiquitin-associated protein 2-like apoptosis proliferation

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触脑脊液神经元的研究进展

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神经解剖学杂志 2018年第5期34卷 633-637页

作者：李瑞刘海鹰王家瑶郭保霖高方武胜昔王文挺空军军医大学基础医学院神经生物学教研室西安710032 空军军医大学基础医学院学员四大队西安710032 空军军医大学基础医学院学员三大队西安710032

触脑脊液神经元（cerebrospinal fluid-contacting neurons,CSF-cNs）是一种分布于脑室、中央管、脑室周器及脑实质等处与脑脊液接触的特殊神经元。根据分布位置不同可将CSF-cNs分为室管膜上、室管膜下和远位CSF-cNs三类,不同部位的CSF-... 详细信息

触脑脊液神经元（cerebrospinal fluid-contacting neurons,CSF-cNs）是一种分布于脑室、中央管、脑室周器及脑实质等处与脑脊液接触的特殊神经元。根据分布位置不同可将CSF-cNs分为室管膜上、室管膜下和远位CSF-cNs三类,不同部位的CSF-cNs分泌不同的神经递质。以往研究CSF-cNs多采用脑室注射辣根过氧化物酶标记的霍乱毒素B亚单位（cholera toxin subunit B labeled with horseradish peroxidase,CB-HRP）进行逆行追踪.

关键词：触脑脊液神经元 Polycystin kidney disease 2-like 1(PKD2L1) Acid-sensing ion channels(ASIC)

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