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Nano-Micro Letters 2020年第6期12卷 113-125页

作者： Yuxin Zhang Xueping Xie Wenjuan Ma Yuxi Zhan Chenchen Mao Xiaoru Shao Yunfeng Lin State Key Laboratory of Oral Diseases National Clinical Research Center for Oral DiseasesWest China Hospital of StomatologySichuan UniversityChengdu 610041People’s Republic of China

Biofilm formation is responsible for numerous chronic infections and represents a serious health challenge.Bacteria and the extracellular polysaccharides(EPS)cause biofilms to become adherent,toxic,resistant to antibiotics,and ultimately difficult to remove.Inhibition of EPS synthesis can prevent the formation of bacterial biofilms,reduce their robustness,and promote removal.Here,we have developed a framework nucleic acid delivery system with a tetrahedral configuration.It can easily access bacterial cells and functions by delivering Antisense oligonucleotides that target specific genes.We designed Antisense oligonucleotide sequences with multiple targets based on conserved regions of the VicK protein-binding site.Once delivered to bacterial cells,they significantly decreased EPS synthesis and biofilm thickness.Compared to existing approaches,this system is highly efficacious because it simultaneously reduces the expression of all targeted genes(gtfBCD,gbpB,ftf).We demonstrate a novel nucleic acid-based nanomaterial with multi-targeted inhibition that has great potential for the treatment of chronic infections caused by biofilms.

关键词： Biofilm Framework nucleic acid Multi-targeting Antisense oligonucleotide Delivery system

Inhibition of allergic airway inflammation by Antisense-induced blockade of STAT6 expression

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Chinese Medical Journal 2011年第1期124卷 26-31页

作者： TIAN Xin-rui TIAN Xin-li BO Jian-ping LI Shao-gang LIU Zhuo-la NIU Bo Department of Respiratory Diseases Second Hospital of Shanxi Medical University Taiyuan Shanxi 030001 China Capital Institute of Pediatrics Beijing 100020 China

Background The signal transducer and activator of transcription 6 （STAT6） expression in lung epithelial cells plays a pivotal role in asthma pathogenesis. Activation of STAT6 expression results in T helper cell type 2 （Th2） cell differentiation leading to Th2-mediated IgE production, development of allergic airway inflammation and hyperreactivity. Therefore, antagonizing the expression and/or the function of STAT6 could be used as a mode of therapy for allergic airway inflammation. Methods In this study, we synthesized a 20-mer phosphorothioate Antisense oligonucleotide （ASODN） overlapping the translation starting site of STAT6 and constructed STAT6 Antisense RNA （pANTI-STAT6）, then transfected them into murine spleen lymphocytes and analyzed the effects of antagonizing STAT6 function in vitro and in a murine model of asthma. Results In vitro, we showed suppression of STAT6 expression and interleukin （IL）-4 production of lymphocytes by STAT6 ASODN. This effect was more prominent when cells were cultured with pANTI-STAT6. In a murine model of asthma associated with allergic pulmonary inflammation in ovalbumin （OVA）-sensitized mice, local intranasal administration of fluorescein isothiocyanate （FITC）-Iabeled STAT6 ASODN to DNA uptake in lung cells was accompanied by a reduction of intracellular STAT6 expression. Such intrapulmonary blockade of STAT6 expression abrogated signs of lung inflammation, infiltration of eosinophils and Th2 cytokine production. Conclusion These data suggest a critical role of STAT6 in the pathogenesis of asthma and the use of local delivery of STAT6 ASODN as a novel approach for the treatment of allergic airway inflammation such as in asthma.

关键词： signal transducer and activator of transcription 6 Antisense asthma lymphocyte Th2 cytokine

汤新景团队在环状反义寡聚核苷酸作为microRNA抑制剂的研究方面取得进展

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Journal of Chinese Pharmaceutical Sciences 2021年第10期30卷 855-856页

作者：北京大学药学院天然药物及仿生药物国家重点实验室北京大学药学院天然药物及仿生药物国家重点实验室

2021年10月21日,北京大学药学院天然药物及仿生药物国家重点实验室汤新景研究团队在学术期刊《Journalof Medicinal Chemistry》发表了最新研究成果"Circular Antisense Oligonucleotides for Specific RNase-H-Mediated micro RNA Inhi... 详细信息

关键词：北京大学药学院反义寡聚核苷酸学术期刊 Antisense 最新研究成果

Effects of long non-coding RNA Opa-interacting protein 5 Antisense RNA 1 on colon cancer cell resistance to oxaliplatin and its regulation of micro RNA-137

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World Journal of Gastroenterology 2020年第13期26卷 1474-1489页

作者： Jing Liang Xiao-Feng Tian Wei Yang Department of Gastrointestinal Surgery China-Japan Union Hospital of Jilin UniversityChangchun 130000Jilin ProvinceChina Department of Hepatopancreatobiliary Surgery China-Japan Union Hospital of Jilin UniversityChangchun 130000Jilin ProvinceChina

BACKGROUND The incidence of colon cancer(CC)is currently high,and is mainly treated with chemotherapy.Oxaliplatin(L-OHP)is a commonly used drug in chemotherapy;however,long-term use can induce drug resistance and seriously affect the prognosis of patients.Therefore,this study investigated the mechanism of Opainteracting protein 5 Antisense RNA 1(OIP5-AS1)on L-OHP resistance by determining the expression of OIP5-AS1 and micro RNA-137(miR-137)in CC cells and the effects on L-OHP resistance,with the goal of identifying new targets for the treatment of CC.AIM To study the effects of long non-coding RNA OIP5-AS1 on L-OHP resistance in CC cell lines and its regulation of miR-137.METHODS A total of 114 CC patients admitted to China-Japan Union Hospital of Jilin University were enrolled,and the expression of miR-137 and OIP5-AS1 in tumor tissues and corresponding normal tumor-adjacent tissues was determined.The influence of OIP5-AS1 and miR-137 on the biological behavior of CC cells was evaluated.Resistance to L-OHP was induced in CC cells,and their activity was determined and evaluated using cell counting kit-8.Flow cytometry was used to analyze the apoptosis rate,Western blot to determine the levels of apoptosisrelated proteins,and dual luciferase reporter assay combined with RNA-binding protein immunoprecipitation to analyze the relationship between OIP5-AS1 and miR-137.RESULTS OIP5-AS1 was up-regulated in CC tissues and cells,while miR-137 was downregulated in CC tissues and cells.OIP5-AS1 was inversely correlated with miR-137(P<0.001).Silencing OIP5-AS1 expression significantly hindered the proliferation,invasion and migration abilities of CC cells and markedly increased the apoptosis rate.Up-regulation of miR-137 expression also suppressed these abilities in CC cells and increased the apoptosis rate.Moreover,silencing OIP5-AS1 and up-regulating miR-137 expression significantly intensified growth inhibition of drug-resistant CC cells and improved the sensitivity of CC cells to LOHP.OIP5-AS1 targetedly inhibited miR-

关键词： Long non-coding RNA Opa-interacting protein 5 Antisense RNA 1 Micro RNA-137 Colon cancer Drug resistance Oxaliplatin Biological behavior

Recombinant adeno-associated virus-mediated delivery of Antisense angiotensin Ⅱ receptor 1 gene attenuates hypertension development

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Acta Pharmacologica Sinica 2007年第11期28卷 1737-1745页

作者： Xu-guang LI~2,Jiang-tao YAN~2,Xi-zheng XU,Jia-ning WANG,Li-ming CHENG,Tao WANG,Ping ZUO,Dao-wen WANG~3Institute of Hypertension and Department of Internal Medicine,Tongji Hospital,Tongji Medical College,Huazhong University of Science andTechnology,Wuhan 430030,China Institute of Hypertension and Department of Internal Medicine Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan China

Aim:The renin-angiotensin system plays a crucial role in the development andestablishment of hypertension,and the pharmacological blockade of the systemresults in a reduction in blood pressure.In the present study,we investigatedwhether the effects of a novel,double-stranded,recombinant adeno-associatedvirus vector (rAAV)-mediated Antisense angiotensin Ⅱ receptor 1 (AT1R) geneefficiently prevents the development of hypertension induced by a high-salt dietin adult,male Sprague-Dawley (SD) rats.Methods:A rAAV was prepared with acassette containing a cytomegalovirus promoter and partial cDNA (660 base pairs)for the AT1R inserted in the Antisense direction (rAAV-AT1-AS).A single tail veininjection of the rAAV-AT1-AS or rAAV-GFP (green fluorescent protein,a reportergene) was performed in adult,male SD rats.Two weeks after injection,the animalswere fed a diet containing 8% NaCl,and the systolic blood pressure was measuredweekly using the tail-cuff method for 12 weeks.Results:The high-salt dietinduced a significant rise in systolic blood pressure in the rAAV-GFP-treatedanimals;however,the rAAV-AT1-AS treatment attenuated the rise in blood pres-sure (142.7±4.5 mmHg vs 117±3.8 mmHg,P<0.01),and the hypotensive effect wasmaintained until the experiments ended at 12 weeks.In the rAAV-GFP-treatedanimals AT1was overexpressed in various tissues,especially in the aorta andkidney at mRNA levels;in contrast,rAAV-AT1-AS treatment markedly attenuatedAT1expression.Furthermore,rAAV-AT1-AS treatment prevented target organdamages from hypertension,including cardiac dysfunction and renal injury com-pared to the rAAV-GFP group.Conclusion:These results suggest that rAAV-mediated anti-AT1delivery attenuates the development of hypertension and pro-tects against renal injury and cardiac remodeling.

关键词： hypertension gene therapy angiotensin Ⅱ receptor 1 Antisense double-stranded recombinant adeno-associated virus vector

Antisense oligonucleotides targeting midkine inhibit tumor growth in an in situ human hepatocellular carcinoma model

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Acta Pharmacologica Sinica 2007年第3期28卷 453-458页

作者： Li-cheng DAI~(2,5) Xiang WANG~2 Xing YAO~3 Li-shan MIN~3 Jin-liang PING~4 Jian-fang HE~2 ~2Huzhou Key Laboratory of Molecular Medicine,~3Department of General Surgery and ~4Department of Pathology Huzhou Central Hospital,Huzhou 313000,China Huzhou Key Laboratory of Molecular Medicine Huzhou Central Hospital Huzhou 313000 China Department of General Surgery Huzhou Central Hospital Huzhou 313000 China Department of Pathology Huzhou Central Hospital Huzhou 313000 China

Aim:To evaluate the in vivo antitumor effects of Antisense oligonucleotides tar-geting midkine(MK-AS).Methods:An in situ human hepatocellular carcinoma(HCC)model was established in mice livers orthotopically.The MK-AS and 5-fluorouracil(5-Fu)were administered intravenously.The tumor sizes and plasmaalpha-fetoprotein(AFP)were measured by calipers and radiation immunoassayrespectively.The morphology of tumors was evaluated by hematoxylin-eosinstaining of histological sections.Human MK,p53,Bax,Bcl-2,and caspase-3protein content were detected by Western blotting.Results:MK-AS signifi-cantly inhibited in situ human HCC growth in mice compared with the saline groupin a dose-dependent manner.After the treatment with MK-AS or with 5-Fu,theplasma AFP concentration decreased in a dose-dependent manner.Interestingly,MK-AS also clearly downregulated the protein level of Bcl-2,and upregulatedp53,Bax,and caspase-3 in the hepatocellular carcinoma tissue.Conclusion:Theseresults demonstrated that MK-AS was an effective antitumor Antisense oligo-nucleotide in vivo in mice;its antitumor effect is associated with the increase ofpro-apoptotic proteins,such as p53,Bax,and caspase-3,and the decrease of theanti-apoptotic protein,Bcl-2.

关键词： oligonucleotides Antisense midkine carcinoma hepatocellular

Inhibition of Inositol-1-phosphate Synthetase in Mycobacterium Tuberculosis by Chitosan-Antisense Nanoparticles

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Journal of Wuhan University of Technology(Materials Science) 2009年第1期24卷 87-90页

作者：李媛媛徐顺清 MOE Key Laboratory School of Public Health Tongji Medical College Huazhong University of Science and Technology

Oligodeoxynucleotides （ODNs） were combined with the biodegradable polymer chitosan to form chitosan-ODN nanoparticles by complex coacervation, in order to improve the sta-bility and intracellular penetration. The diameter of the nanoparticles was light strength size and ranged between 60 and 219 nm with a mean value of 132 nm, while zeta potential was between ＋12 and ＋20 mV at pH 5.5. The chitosan-ODN nanoparticles could partially protect the encapsulated ODN from nuclease degradation. Moreover, chitosan-ODN nanoparticles were much more effective in inhibiting the proliferation of M.tuberculosis than free ODN.

关键词： chitosan Antisense oligodeoxynucleotides nanoparticle tuberculosis

Effects of a plasmid expressing Antisense tissue inhibitor of metalloproteinase-1 on liver fibrosis in rats

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Chinese Medical Journal 2005年第3期118卷 192-197页

作者： JIANGWei WANGJi-yao YANGChang-qing LIUWen-bin WANGYi-qing HEBo-ming DepartmentofGastroenterology ZhongshanHospitalFudanUniversityShanghai200032China

No efficient therapy for liver fibrosis has been available. This study wasaimed to provide evidence that the introduction of a plasmid expressing Antisense tissue inhibitorof metalloproteinase-1 (TIMP-1) into a rat model of immunologically induced liver fibrosis canresult in the increased activity of interstitial collagenase, thus enhancing the degradation ofcollagen. Real-time nested polymerase chain reaction ( RT-Nested-PCR) and gene recombinationtechniques were used to construct a rat Antisense TIMP-1 recombinant plasmid that can be expressedin eukaryotic cells. Both the recombinant plasmid and an empty vector ( _(pc)DNA3 ) wereencapsulated with glycosyl-poly-L-lysine and injected into rats suffering from pig serum-inducedliver fibrosis. The expression of ex'ogenous transfected plasmid was assessed by Northern blot,RT-PCR, and Western blot. Hepatic interstitial collagenase activity was detected usingfluorescinisothiocyanate ( FITC)-labeledtype Ⅰ collagen. In addition to hepatic hydroxyprolinecontent, hepatic collagen types Ⅰ and Ⅲ were detected by immunohistochemical staining, and thestages of liver fibrosis by Van Gieson staining. Exogenous Antisense TIMP-1 was successfullyexpressed in vivo and could block the gene and protein expression of TIMP-1. Active and latenthepatic interstitial collagenase activities were elevated (PAntisense TIMP-1 group (P Antisense TIMP-1 recombinant plasmids have some inhibitory effect on liver fibrosis.

关键词： liver fibrosis Antisense tissue-inhibitor of metalloproteinase-1

Antisense expression of a rice cellular apoptosis susceptibility gene (OsCAS} alters the height of transgenic rice

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Progress in Natural Science:Materials International 2007年第1期17卷 39-43页

作者： XU Chunxiao and HE Chaozu ( National Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China Graduate University of Chinese Academy of Sciences, Beijing 100039, China Shenzhen Laboratory of Superhybrid Rice Research, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China) National Key Laboratory of Plant Genomics Institute of Microbiology Chinese Academy of Sciences Beijing 100080 China Graduate University Chinese Academy of Sciences Beijing 100039 China Shenzhen Laboratory of Superhybrid Rice Research Graduate School at Shenzhen Tsinghua University Shenzhen 518055 China

Cellular apoptosis susceptibility (CAS) gene plays important roles in mitosis, development and export of importin a from the nucleus, but its function in plant is unknown. In this study, a rice CAS ortholog (OsCAS), which encodes a predicted protein of 983 amino acids with 62% similarity to human CAS, was identified. DNA gel blot analysis revealed a single copy of OsCAS in the rice genome. A 973 bp fragment at the 3’ end of OsCAS cDNA was cloned from rice cDNA library and transferred into rice in the Antisense direction under the control of CaMV 35S promoter via Agrobacterium-mediated transformation method, 105 transgenic lines were obtained. Expression of OsCAS was suppressed in the Antisense transgenic lines as revealed by semi-quantitative RT-PCR. The Antisense transgenic lines showed dwarf phenotypes. The results indicated that OsCAS was involved in culm development of rice.

关键词： Antisense dwarf ortholog OsCAS rice.

Evidence for the Antisense transcription in the proviral R29-127 strain of bovine immunodeficiency virus

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Virologica Sinica 2015年第3期30卷 224-227页

作者： Bin Liu Xuechao Zhao Wenyuan Shen Xiaohong Kong Laboratory of Medical Molecular Virology School of Medicine Nankai University Tianjin 300071 China

Dear Editor,In the late stages of the retroviral life cycle,the transcription of viral pre-m RNA is initiated at the 5′long terminal repeat(5′LTR)and terminated at the 3′LTR;the full-length and spliced viral RNAs are transported out of the nucleus and serve as templates for the translation of viral proteins or alternatively as the full-length viral genome to be packaged into virus particles.

关键词： Antisense terminated transcripts nucleus initiated length primer plasmid bovine transfection