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Inhibition of Proteasome LMP2 Activity Suppresses Chil3 Expression in Mouse Colon Adenocarcinoma Tissue and Restrains Tumor Growth
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作者 Tatiana M.Astakhova Nikita S.Karpov +7 位作者 Nataliya O.Dashenkova Elena V.Alpeeva Mikhail V.Nesterchuk Sergey B.Akopov Arsen S.Mikaelyan Anfisa S.Ryabchenko Pavel A.Erokhov Natalia P.Sharova 《Oncology Research》 2025年第9期2573-2595,共23页
Objectives:Proteasomes,multi-subunit proteases,are key actors of cellular protein catabolism and a number of regulatory processes.The detection of subtle proteasome functioning in tumors may contribute to our understa... Objectives:Proteasomes,multi-subunit proteases,are key actors of cellular protein catabolism and a number of regulatory processes.The detection of subtle proteasome functioning in tumors may contribute to our understanding of the mechanisms of cancer development.The current study aimed to identify the role of low molecular mass protein 2(LMP2),a proteasome immune subunit,in the development of mouse colon 26(C26)adenocarcinoma.Methods:The functions of the LMP2 subunit in tumor development in Balb/c mice were studied using its irreversible inhibitor KZR-504.LMP2 activity was detected by the hydrolysis of the fluorogenic substrate Ac-Pro-Ala-Leu-AMC.Western blotting and Quantitative Reverse Transcription Polymerase Chain Reaction(qRT-PCR)were used.We applied fluorescent tests for cell proliferation and apoptosis.M2 macrophages were obtained by polarization of mouse bone marrow-derived macrophages using the corresponding cytokines.Results:KZR-504 showed high specificity only for the LMP2 subunit and had no negative effect on C26 cells in culture.However,KZR-504 suppressed the formation of tumor conglomerates(by 74%,p<0.001)after C26 cell transplantation in vivo,inhibited the expression of chitinase-<3-like protein 3(Chil3)gene(by 90%,p<0.001),a key marker of immunosuppressive M2 macrophages,in the tumor<microenvironment,and reduced the tumor weight compared to the control(by 48%,p<0.01).KZR-504 also suppressed<the expression of Chil3(by 68%,p<0.05)and arginase-1(Arg1)(by 90%,p<0.001),another marker gene,in M2<<macrophages and violated M0-M2 macrophage polarization in culture.Conclusion:We discovered earlier unknown functions of the proteasome LMP2 subunit to facilitate the formation of tumor conglomerates and maintain Chil3 and Arg1 expression in immunosuppressive M2 macrophages.Our work demonstrates that the proteasome LMP2 subunit can be a target for antitumor treatment. 展开更多
关键词 mouse colon 26 adenocarcinoma M2 macrophages proteasome low molecular mass protein 2 subunit chitinase-3-like protein 3 KZR-504
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SOCS3 Expression Correlates with Severity of Inflammation in Mouse Hepatitis Virus Strain 3-induced Acute Liver Failure and HBV-ACLF 被引量:9
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作者 李咏 韩梅芳 +11 位作者 李维娜 师爱超 张元亚 王宏艳 王发席 李兰 吴婷 丁琳 陈韬 严伟明 罗小平 宁琴 《Journal of Huazhong University of Science and Technology(Medical Sciences)》 SCIE CAS 2014年第3期348-353,共6页
Summary: Recently, suppressor of cytokine signaling-3 (SOCS3) has been shown to be an inducible endogenous negative regulator of Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway ... Summary: Recently, suppressor of cytokine signaling-3 (SOCS3) has been shown to be an inducible endogenous negative regulator of Janus kinase/signal transducers and activators of transcription (JAK/STAT) pathway which is relevant in inflammatory response, while its functions in acute liver failure and HBV-induced acute-on-chronic liver failure (HBV-ACLF) have not been fully elucidated. In this study, we explored the role of SOCS3 in the development of mouse hepatitis virus strain 3 (MHV-3)-induced acute liver failure and its expression in liver and peripheral blood mononuclear cells (PBMCs) of patients with HBV-ACLF. Inflammation-related gene expression was detected by real-time PCR, immtmohistochemistry and Western blotting. The correlation between SOCS3 level and liver injury was studied. Our results showed that the SOCS3 expression was significantly elevated in both the liver tissue and PBMCs from patients with HBV-ACLF compared to mild chronic hepatitis B (CHB). Moreover, a time course study showed that SOCS3 level was increased remarkably in the liver of BALB/cJ mice at 72 h post-infection. Pro-inflammatory cytokines, interleukin (IL)-1 β, IL-6, and tumor necrosis factor (TNF)-α, were also increased significantly at 72 h post-infection. There was a close correlation between hepatic SOCS3 level and IL-6, and the severity of liver injury defined by alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, respectively. These data suggested that SOCS3 may play a pivotal role in the pathogenesis of MHV-3-induced acute liver failure and HBV-ACLF. 展开更多
关键词 suppressors of cytokine signaling-3 HBV-induced acute-on-chronic liver failure mouse hepatitis virus strain 3 fulminant liver failure BALB/cJ mice
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Automated Behavioral Phenotyping Reveals Presymptomatic Alterations in a SCA3 Genetrap Mouse Model 被引量:1
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作者 Jeannette Hübener Nicolas Casadei +5 位作者 Peter Teismann Mathias W. Seeliger Maria Bjrkqvist Stephan von Horsten Olaf Riess Huu Phuc Nguyen 《Journal of Genetics and Genomics》 SCIE CAS CSCD 2012年第6期287-299,共13页
Characterization of disease models of neurodegenerative disorders requires a systematic and comprehensive phenotyping in a highly standardized manner. Therefore, automated high-resolution behavior test systems such as... Characterization of disease models of neurodegenerative disorders requires a systematic and comprehensive phenotyping in a highly standardized manner. Therefore, automated high-resolution behavior test systems such as the homecage based LabMaster system are of particular interest. We demonstrate the power of the automated LabMaster system by discovering previously unrecognized features of a recently characterized atxn3 mutant mouse model. This model provided neurological symptoms including gait ataxia, tremor, weight loss and premature death at the age of 12 months usually detectable just 2 weeks before the mice died. Moreover, using the LabMaster system we were able to detect hypoactivity in presymptomatic mutant mice in the dark as well as light phase. Additionally, we analyzed inflammation, immunological and hematological parameters, which indicated a reduced immune defense in phenotypic mice. Here we demonstrate that a detailed characterization even of organ systems that are usually not affected in SCA3 is important for further studies of pathogenesis and required for the preclinical therapeutic studies. 展开更多
关键词 Automated homecage behavior Genetrap mouse model Spinocerebellar Ataxia Type 3
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Transglutaminase 3 expression in C57BL/6J mouse embryo epidermis and the correlation with its differentiation 被引量:3
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作者 JianZHANG HuiYingZHI +2 位作者 FangDING AiPingLUO ZhiHuaLIU 《Cell Research》 SCIE CAS CSCD 2005年第2期105-110,共6页
Epidermal-type transglutaminase 3 (TGM3) is involved in the cross-linking of structural proteins to form the cornifiedenvelope in the epidermis. In the present study, we detected the expression of TGM3 in the mouse em... Epidermal-type transglutaminase 3 (TGM3) is involved in the cross-linking of structural proteins to form the cornifiedenvelope in the epidermis. In the present study, we detected the expression of TGM3 in the mouse embryo using RT-PCR.TGM3 mRNA is weakly presented from E11.5 to E14.5 and increases significantly from E15.5 to birth. Then wedetermined the spatial and temporal expression pattern of TGM3 in the skin and other organs by in situ hybridization. Wefound a deprivation of TGM3 in skin at E11.5, while a rich supply in periderm cells and a weak expression in basal cellsfrom E12.5 to E14.5. From the period of E15.5 to E16.5, after keratinization in the epidermis, TGM3 was expressed inthe granular and cornified layers. The electron microscopic observation of the C57BL/6J mouse limb bud skin develop-ment provided several morphological evidences for the epidermal differentiation. The above findings suggest that theexpression of TGM3 plays a important role in the epidermis differentiation in embryogenesis. 展开更多
关键词 transglutaminase 3 EPIDERMIS DIFFERENTIATION C57BL/6J mouse embryo.
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Cytotoxicity of seven recent dentine bonding agents on mouse 3T3 fibroblast cells 被引量:3
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作者 Annette Olivier Sias R. Grobler Yusuf Osman 《Open Journal of Stomatology》 2012年第4期244-250,共7页
Today it is generally accepted that most bonding agents are cytotoxic. In this study the relative cytotoxicity of seven recent dentine bonding agents on mouse 3T3 fibroblast cells were investigated. Materials and Meth... Today it is generally accepted that most bonding agents are cytotoxic. In this study the relative cytotoxicity of seven recent dentine bonding agents on mouse 3T3 fibroblast cells were investigated. Materials and Methods. Near-confluent mouse 3T3 fibroblast cells were exposed to Dulbecco Modified Eagle’s Medium containing extractions from the seven different bonding agents. The cell survival rate was then determined using the standard MTT assay. Results. The cell survival rate ranking is: iBond (94%) < Gbond (78%) < Xeno V (71%) < Adper Easy Bond (63%) < Xeno V+ (61%) < Adper Scotchbond SE (33%) < XP Bond (32%). Part A of Adper Scotchbond SE had a survival rate of 35% and part B 38%. These two parts did not differ significantly. Adper Scotchbond SE and XP Bond do not differ significantly. While Xeno V+, Xeno V and Adper Easy Bond do not differ. (p < 5%;Tukey-Kramer Multiple-Comparison Test). Conclusion. All of the tested adhesive bonding agents were cytotoxic with survival rate of 3T3 cells between 94% to 31%. Of the 7 bonding agents tested iBond was found to be only slightly toxic and by far the least toxic. The two bonding agents (XP Bond and Adper Scotchbond SE) containing UDMA plus TEGDMA plus HEMA plus camphorquinone were found to be the most toxic. 展开更多
关键词 CYTOTOXICITY BONDING AGENTS mouse 3T3 FIBROBLAST
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A Three-Dimensional (3D) Environment to Maintain the Integrity of Mouse Testicular Can Cause the Occurrence of Meiosis 被引量:1
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作者 CHU Zhi-li LIU Chao +3 位作者 BAI Yao-fu ZHU Hai-jing HU Yue HUA Jin-lian 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第8期1481-1488,共8页
Adhesions between different cells and extracellular matrix have been studied extensively in vitro, but little is known about their functions in testicular tissue counterparts. Spermatogonia and their companion somatic... Adhesions between different cells and extracellular matrix have been studied extensively in vitro, but little is known about their functions in testicular tissue counterparts. Spermatogonia and their companion somatic cells maintain a close association throughout spermatogenesis and this association is necessary for normal spermatogenesis. In order to keep the relative integrity of the testicular tissues, and to detect the development in vitro, culture testicular tissues in a three- dimensional (3D) agarose matrix was examined. Testicular tissues isolated from 6.5 d postpartum (dpp) mouse were cultured on the top of the matrix for 26 d with a medium height up to 4/5 of the 3D agarose matrix. The results showed that in this 3D culture environment, each type of testicular cells kept the same structure, localization and function as in vivo and might be more biologically relevant to living organisms. After culture, germ cell marker VASA and meiosis markers DAZL and SCP3 showed typical positive analysed by immunofluorescence staining and RT-PCR. It demonstrated that this 3D culture system was able to maintain the number of germ cells and promote the meiosis initiation of male germ cells. 展开更多
关键词 three-dimensional culture 3D) MEIOSIS organ culture mouse
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Changes of the intestinal endocrine cells in the C57BL/6 mouse after implantation of murine lung carcinoma (3LL): An immunohistochemical quantitative study
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作者 Sae-Kwang Ku Seung-Kyoo Seong +5 位作者 Dae-Young Kim Hyeung-Sik Lee Jong-Dae Kim Hae-Yun Choi Bu-Il Seo Jae-Hyun Lee 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第35期5460-5467,共8页
AIM: To study the distributions and frequencies of intestinal endocrine cells in the C57BL/6 mouse with immunohistochemical method using seven types of specific antisera against chromogranin A (CGA), serotonin,somatos... AIM: To study the distributions and frequencies of intestinal endocrine cells in the C57BL/6 mouse with immunohistochemical method using seven types of specific antisera against chromogranin A (CGA), serotonin,somatostatin, glucagons, gastrin, cholecystokinin (CCK)-8 and human pancreatic polypeptide (hPP) after abdominal subcutaneous implantation of murine lung carcinoma (3LL).METHODS: The experimental animals were divided into two groups, one is non-implanted Sham and the other is 3LL-implanted group. Samples were collected from six regions of intestinal tract at 28th d after implantation of 3LL cells (1×105 cell/mouse).RESULTS: In this study, five types of immunoreactive (IR) cells were identified except for gastrin and hPP. The regional distributions of the intestinal endocrine cells in the 3LL-implanted group were similar to those of the non-implanted Sham. However, significant decreases of IR cells were detected in 3LL-implanted group compared to those of non-implanted Sham. CGA- and serotonin-IR cells significantly decreased in 3LL-implanted groups compared to that of non-implanted Sham. Somatostatin-IR cells in the jejunum and ileum and CCK-8-IR cells in the jejunum of 3LL-implanted groups significantly decreased compared to that of non-implanted Sham. In addition,glucagon-IR cells were restricted to the ileum and colon of non-implanted Sham.CONCLUSION: Implantation of tumor cell mass (3LL)induced severe quantifiable changes of intestinal endocrine cell density and the abnormality in density of intestinal endocrine cells may contribute to the development of gastrointestinal symptoms such as anorexia and indigestion, frequently encountered in patients with cancer. 展开更多
关键词 Intestinal endocrine cell IMMUNOHISTOCHEMISTRY C57BL/6 mouse 3LL Tumor
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N-methyl-D-aspartate receptor subtype 3A promotes apoptosis in developing mouse brain exposed to hyperoxia
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作者 Jimei Li Shanping Yu +2 位作者 Zhongyang Lu Osama Mohamad Ling Wei 《Neural Regeneration Research》 SCIE CAS CSCD 2012年第4期273-277,共5页
In the present study, 7 day postnatal C57/BL6 wild-type mice (hyperoxia group) and 7 day postnatal N-methyI-D-aspartate receptor subtype 3A knockout mice (NR3A KO group) were exposed to 75% oxygen and 15% nitrogen... In the present study, 7 day postnatal C57/BL6 wild-type mice (hyperoxia group) and 7 day postnatal N-methyI-D-aspartate receptor subtype 3A knockout mice (NR3A KO group) were exposed to 75% oxygen and 15% nitrogen in a closed container for 5 days. Wild-type mice raised in normoxia served as controls. TdT-mediated dUTP nick end labeling (TUNEL)/neuron-specific nuclear protein (NeuN) and 5-bromo-2'-deoxyuridine (BrdU)/NeuN immunofluorescence staining showed that the number of apoptotic cells and the number of proliferative cells in the dentate subgranular zone significantly increased in the hyperoxia group compared with the control group. However, in the same hyperoxia environment, the number of apoptotic cells and the number of proliferative cells significantly decreased in the NR3A KO group compared with hyperoxia group. TUNEL+/NeuN+ and BrdU+/NeuN~ cells were observed in the NR3A KO and the hyperoxia groups. These results demonstrated that the NR3A gene can promote cell apoptosis and mediate the potential damage in the developing brain induced by exposure to non-physiologically high concentrations of oxygen. 展开更多
关键词 N-methyl-D-aspartate receptor subtype 3A apoptosis cell proliferation HYPEROXIA developing brain nerve cells mouse NEUROBIOLOGY neural regeneration
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Effects of DHRS3 in C2C12 Myoblast Differentiation and Mouse Skeletal Muscle Injury
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作者 Zhang Wen-yu Xu Jia-hui +3 位作者 Zhang Chun-yu Tong Hui-li Li Shu-feng Yan Yun-qin 《Journal of Northeast Agricultural University(English Edition)》 CAS 2021年第3期38-47,共10页
Myoblast differentiation is an essential process during skeletal muscle development.C2C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro.Dehydrogenase/reductase(SDR family... Myoblast differentiation is an essential process during skeletal muscle development.C2C12 myoblast is a commonly used experimental model to study muscle cell differentiation in vitro.Dehydrogenase/reductase(SDR family)member 3(DHRS3)is a highly conserved member in short-chain alcohol dehydrogenase/reductase superfamily and has been shown to be involved in the metabolism of retinol.Previous experimental results showed that the expression of DHRS3 increased significantly during the differentiation of myoblasts differentiation.However,the effect of DHRS3 on mouse muscle cell differentiation was unclear.The objective of current study was to determine if DHRS3 affected muscle cell differentiation,and if DHRS3 was involved in muscle regeneration.Protein expression was determined by western blot and immunofluorescence analysis.The activation and inhibition of DHRS3 increased and decreased C2C12 myoblast differentiation respectively,which indicated that DHRS3 could affect C2C12 myoblast differentiation.DHRS3 expression was significantly changed during muscle regeneration,with the regeneration of muscle injury,the expression of DHRS3 tended to increase first and then decrease.It suggested that DHRS3 might be involved in muscle regeneration.In summary,this study confirmed the involvement of DHRS3 in C2C12 myoblast differentiation and mouse skeletal muscle regeneration and provided a theoretical basis for further elucidating the molecular mechanism of muscle development. 展开更多
关键词 DHRS3 C2C12 cell differentiation mouse skeletal muscle injury
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Kanglaite combined Gemcitabine inhibits growth of nude mouse subcutaneous transplantation tumor of human PC-3 pancreatic cancer cell
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作者 王伟 金建光 秦兆寅 《Journal of Medical Colleges of PLA(China)》 CAS 2005年第4期219-222,235,共5页
Objective:To study the mechanisms of pancreatic cancer treatment with Kanglaite combined Gemcitabine by investigating the relationship between the apoptosis and the expression of bcl-2, Bax and VEGF in pancreatic canc... Objective:To study the mechanisms of pancreatic cancer treatment with Kanglaite combined Gemcitabine by investigating the relationship between the apoptosis and the expression of bcl-2, Bax and VEGF in pancreatic cancer cells.Methods:Nude mouse subcutaneous transplantation tumor model of Human PC-3 pancreatic cancer was established; the expressions of bcl-2, Bax and VEGF of transplantation tumor cell were determined; the earlier apoptosis rate of pancreatic cancer cell and the gross tumor volume were determined. Results:Kanglaite combined Gemcitabine remarkably decreased the protein expression of bcl-2,raised the expression of Bax,increased the apoptosis rate of the pancreatic cancer and contract the gross tumor volume. Kanglaite greatly decreased the protein expression of VEGF of the tumor cell. Conclusion:Therapeutic efficacy of Kanglaite combined Gemcitabine is far better than separate use of the two medicines in the pancreatic cancer transplantation tumor treatment. 展开更多
关键词 human PC-3 pancreatic cancer nude mouse subcutaneous transplantation tumor apoptosis immunohistochemisry bcl-2 Bax VEGF
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Application of Novel Rotation Angular Model for 3D Mouse System Based on MEMS Accelerometers
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作者 钱莉 陈文元 徐国平 《Journal of Shanghai Jiaotong university(Science)》 EI 2009年第2期137-142,共6页
A new scheme is proposed to model 3D angular motion of a revolving regular object with miniature, low-cost micro electro mechanical systems(MEMS) accelerometers(instead of gyroscope),which is employed in 3D mouse syst... A new scheme is proposed to model 3D angular motion of a revolving regular object with miniature, low-cost micro electro mechanical systems(MEMS) accelerometers(instead of gyroscope),which is employed in 3D mouse system.To sense 3D angular motion,the static property of MEMS accelerometer,sensitive to gravity acceleration,is exploited.With the three outputs of configured accelerometers,the proposed model is implemented to get the rotary motion of the rigid object.In order to validate the effectiveness of the proposed model,an input device is developed with the configuration of the scheme.Experimental results show that a simulated 3D cube can accurately track the rotation of the input device.The result indicates the feasibility and effectiveness of the proposed model in the 3D mouse system. 展开更多
关键词 micro electro mechanical systems (MEMS) accelerometer 3D mouse rotation angular model homogeneous transformation matrix 3D graphical user interface
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In vitro responses of human pulp cells and 3T3 mouse fibroblasts to six contemporary dental restoratives
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作者 Jun Sun Yiming Weng +1 位作者 Fengyu Song Dong Xie 《Journal of Biomedical Science and Engineering》 2011年第1期18-28,共11页
In vitro responses of human primary pulp cells (HPCs) and 3T3 mouse fibroblasts to six contempo-rary commercial dental restoratives were evaluated using the WST-1 assay. The results show that Fuji II is not cytotoxic ... In vitro responses of human primary pulp cells (HPCs) and 3T3 mouse fibroblasts to six contempo-rary commercial dental restoratives were evaluated using the WST-1 assay. The results show that Fuji II is not cytotoxic to both cells. Fuji II LC is not cyto-toxic to HPCs but cytotoxic to 3T3 cells, indicating that 3T3 cells are more vulnerable to 2-hydroxyethyl methacrylate (HEMA) than HPCs. Vitremer is very cytotoxic probably due to having diphenyliodonium chloride and HEMA in it. Z100 is very cytotoxic probably due to having triethylene glycol dimethacry-late (TEGDMA) in it. P60 is cytotoxic but less cyto-toxic than Z100 probably due to no TEGDMA in it. Durelon is the most cytotoxic among the six materials studied probably due to the high cytotoxicity of zinc ions. Additionally, the cytotoxcity of the tested mate-rials was found to be dose-dependent. 展开更多
关键词 In Vitro Cytotoxicity HUMAN Pulp CELLS 3T3 mouse Fibroblast CELLS DENTAL CEMENT GLASS-IONOMER CEMENT Resin Composite
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Lack of an Additive Effect between the Deletions of Klf5 and Nkx3-1 in Mouse Prostatic Tumorigenesis
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作者 Changsheng Xing Xiaoying Fu +1 位作者 Xiaodong Sun Jin-Tang Dong 《Journal of Genetics and Genomics》 SCIE CAS CSCD 2013年第6期315-318,共4页
Prostate cancer is one of the most common malignancies.The development and progression of prostate cancer are driven by a series of genetic and epigenetic events including gene amplification that activates oncogenes a... Prostate cancer is one of the most common malignancies.The development and progression of prostate cancer are driven by a series of genetic and epigenetic events including gene amplification that activates oncogenes and chromosomal deletion that inactivates tumor suppressor genes.Whereas gene amplification occurs in human prostate cancer,gene deletion is more common, 展开更多
关键词 Lack of an Additive Effect between the Deletions of Klf5 and Nkx3-1 in mouse Prostatic Tumorigenesis
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非小细胞肺癌中PRL-3与RhoC的表达及相关性意义 被引量:10
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作者 张平 张志培 +4 位作者 李香敏 雷杰 苏凯 李小飞 王小平 《中国肺癌杂志》 CAS 2010年第6期598-601,共4页
背景与目的PRL-3是新近发现的酪氨酸蛋白磷酸酶,尚属于肝再生磷酸酶家族,具有促进肿瘤转移作用;RhoC属于小分子G蛋白超家族中的Rho亚家族,二者作用机制不清楚。本研究通过检测非小细胞肺癌(non-small celllung cancer,NSCLC)中PRL-3和R... 背景与目的PRL-3是新近发现的酪氨酸蛋白磷酸酶,尚属于肝再生磷酸酶家族,具有促进肿瘤转移作用;RhoC属于小分子G蛋白超家族中的Rho亚家族,二者作用机制不清楚。本研究通过检测非小细胞肺癌(non-small celllung cancer,NSCLC)中PRL-3和RhoC的表达,分析二者表达的相关性及在不同分组之间的差异,为进一步研究PRL-3在肿瘤发生发展中的作用机理提供实验依据。方法采用免疫组化SP法检测PRL-3和RhoC在92例NSCLC中的表达,用统计学方法检验分析二者在不同分组间的表达差异性及二者的相关性。结果在NSCLC中PRL-3和RhoC的表达阳性率分别为69.6%(64/92)、73.9%(68/92),二者的表达在不同的TNM分期、淋巴结及胸膜是否转移的分组之间差异有统计学意义(P<0.01),同时二者的表达具有相关性(r=0.754,P<0.001)。结论在NSCLC中PRL-3和RhoC在TNM分期较高以及合并淋巴结、胸膜转移的分组中表达较高,同时二者的表达具有相关性,提示PRL-3可能通过RhoC及其下游因子促进癌细胞的远处转移。 展开更多
关键词 肺肿瘤 prl-3 RHOC 免疫组织化学
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PRL-3基因RNA干扰慢病毒载体的构建及在SW480细胞中的表达 被引量:6
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作者 柳玉红 李建明 +1 位作者 周军 丁彦青 《南方医科大学学报》 CAS CSCD 北大核心 2008年第4期509-512,共4页
目的构建PRL-3基因慢病毒干扰载体及建立PRL-3基因稳定干扰的结直肠癌细胞亚系,为PRL-3基因功能研究奠定基础。方法设计PRL-3基因特异性RNAi靶序列,与pENTRTM/U6线性载体定向连接,构建pENTRTM/U6真核入门表达载体。通过与pENTRTM/U6 ent... 目的构建PRL-3基因慢病毒干扰载体及建立PRL-3基因稳定干扰的结直肠癌细胞亚系,为PRL-3基因功能研究奠定基础。方法设计PRL-3基因特异性RNAi靶序列,与pENTRTM/U6线性载体定向连接,构建pENTRTM/U6真核入门表达载体。通过与pENTRTM/U6 entry clone与pLenti6/BLOCK-iTTM-DEST载体进行重组,获得pLenti6/BLOCK-iTTM-DEST真核表达慢病毒干扰载体。利用包装细胞293FT获得重组的慢病毒,感染大肠癌细胞株SW480细胞,杀稻瘟菌素筛选获得稳定干扰PRL-3基因的细胞亚系。结果成功构建PRL-3pLenti6/BLOCK-iTTM-DEST真核表达慢病毒干扰载体并获得相应的慢病毒,病毒悬液的滴度为6×105U/L。RT-PCR、Western blotting结果表明,克隆1PRL-3 mRNA及蛋白显著降低。结论成功构建人PRL-3基因特异性的慢病毒干扰载体,获得PRL-3基因稳定干扰的大肠癌细胞亚系。 展开更多
关键词 RNA干扰 慢病毒 prl-3 结直肠癌
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石斛酚通过NF-κB/PRL-3通路抑制骨肉瘤细胞增殖、迁移和侵袭 被引量:11
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作者 范兆阳 鲜文峰 +1 位作者 刘永喜 张超 《中国肿瘤生物治疗杂志》 CAS CSCD 北大核心 2019年第10期1095-1100,共6页
目的:探讨石斛酚对人成骨肉瘤U20S细胞增殖、迁移及侵袭的抑制作用及其分子机制。方法:用不同质量浓度(10、25、50、75、100、150μmol/L)的石斛酚处理U20S细胞24、48h后,用CCK-8法检测石斛酚对U20S细胞增殖能力的影响。Transwell实验检... 目的:探讨石斛酚对人成骨肉瘤U20S细胞增殖、迁移及侵袭的抑制作用及其分子机制。方法:用不同质量浓度(10、25、50、75、100、150μmol/L)的石斛酚处理U20S细胞24、48h后,用CCK-8法检测石斛酚对U20S细胞增殖能力的影响。Transwell实验检测25、50μmol/L的石斛酚对U20S细胞迁移、侵袭能力的影响,在脂多糖(LPS)诱导U20S细胞炎症反应的基础上,再以石斛酚处理,qPCR、WB法分别检测U20S细胞中NF-κB(p65)、TNF-α、IL-6、PRL-3mRNA和蛋白的表达水平。结果:不同质量浓度的石斛酚作用不同时间对肉瘤细胞U20S增殖均具有抑制作用(P<0.05或P<0.01);25、50μmol/L的石斛酚能够明显抑制骨肉瘤U20S细胞的迁移及侵袭能力(均P<0.01),同时能够抑制细胞中NF-κB、TNF-α、IL-6、PRL-3等蛋白的表达(P<0.05或P<0.01);LPS诱导后,U20S细胞中NF-κB、TNF-α、IL-6、PRL-3mRNA和蛋白表达水平均显著升高(均P<0.01),25、50μmol/L的石斛酚处理后均能够降低U20S细胞中NF-κB、TNF-α、IL-6、PRL-3mRNA和蛋白的表达水平(P<0.05或P<0.01)。结论:石斛酚对骨肉瘤U20S细胞增殖、迁移和侵袭具有抑制作用,其作用机制可能与调控NF-κB/PRL-3信号通路有关。 展开更多
关键词 石斛酚 骨肉瘤 U20S细胞 增殖 侵袭 迁移 NF-ΚB prl-3
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PRL-3在前列腺癌组织中的表达及其意义 被引量:8
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作者 赵易 武国军 +5 位作者 张瑞 沈岚 于垂恭 王映梅 药立波 袁建林 《实用医学杂志》 CAS 北大核心 2010年第1期39-41,共3页
目的:研究蛋白酪氨酸磷酸酶PRL-3在前列腺癌及前列腺癌旁增生组织中的表达及其与临床病理特征和生物学行为的关系。方法:采用免疫组织化学S-P法检测60例前列腺石蜡包埋组织中PRL-3的表达情况,其中前列腺增生组织12例,前列腺癌组织48例,... 目的:研究蛋白酪氨酸磷酸酶PRL-3在前列腺癌及前列腺癌旁增生组织中的表达及其与临床病理特征和生物学行为的关系。方法:采用免疫组织化学S-P法检测60例前列腺石蜡包埋组织中PRL-3的表达情况,其中前列腺增生组织12例,前列腺癌组织48例,均为存档石蜡切片。并应用χ2检验方法分析PRL-3在前列腺癌组织中表达的意义与临床病理特征之间的关系。结果:免疫组化结果显示PRL-3表达定位于前列腺癌组织的细胞浆,癌组织与癌旁增生组织的阳性率为分别为79.2%和25.0%(P<0.05),癌组织中PRL-3的表达明显高于癌旁增生组织。PRL-3表达水平在不同年龄段、有无家族史之间差异无显著性(P>0.05),在不同Gleason评分之间差异有显著性(P<0.05),在不同临床TNM分期之间差异无显著性(P>0.05),在血清PSA浓度≤20ng/mL与PSA浓度>20ng/mL组间差异无显著性(P>0.05)。结论:PRL-3的表达与前列腺癌发生发展机制关系较密切,PRL-3蛋白可作为前列腺癌诊断、治疗及预后判断过程中的一种较有价值的病理学指标。 展开更多
关键词 前列腺肿瘤 prl-3基因 良性前列腺增生 免疫组织化学
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miR495、miR551a靶向干扰SGC7901细胞PRL-3基因的表达 被引量:4
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作者 蒋蒙蒙 曹毅 +1 位作者 李正荣 揭志刚 《世界华人消化杂志》 CAS 北大核心 2012年第34期3361-3365,共5页
目的:构建靶向PRL-3基因的miRNA495和miRNA551a真核表达载体,观察其转染胃癌SGC7901细胞后对胃癌细胞PRL-3基因表达的影响.方法:设计并合成两条针对PRL-3基因的特异性miRNA495和miRNA551a干扰序列,分别与pcDNA6.2-GW/Em GFP-miR载体连接... 目的:构建靶向PRL-3基因的miRNA495和miRNA551a真核表达载体,观察其转染胃癌SGC7901细胞后对胃癌细胞PRL-3基因表达的影响.方法:设计并合成两条针对PRL-3基因的特异性miRNA495和miRNA551a干扰序列,分别与pcDNA6.2-GW/Em GFP-miR载体连接,转化大肠杆菌,纯化并鉴定后利用LipofectamineTM2000转染胃癌SGC7901细胞,实时定量PCR及Western blot技术鉴定重组体对PRL-3基因表达的干扰效果.结果:针对PRL-3基因的miRNA495和miRNA551a干扰质粒构建成功,胃癌SGC7901细胞分别转染两种质粒后miRNA495及miRNA551a的表达明显升高,并且明显抑制了PRL-3基因mRNA及蛋白的表达.结论:靶向PRL-3基因的miRNA495和miRNA551a真核表达载体构建成功,其可有效提高胃癌SGC7901细胞miRNA495和miRNA551a的表达,并抑制PRL-3mRNA及PRL-3蛋白的表达. 展开更多
关键词 prl-3 miRNA495 miRNA551a RNA干扰 胃癌
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PRL-3和RhoC在A549细胞中的表达及意义 被引量:3
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作者 张平 张志培 +4 位作者 李香敏 雷杰 苏凯 李小飞 王小平 《中国肺癌杂志》 CAS 2010年第12期1160-1164,共5页
背景与目的在前期研究中,我们发现在非小细胞肺癌中PRL-3和RhoC的表达具有相关性,提示两者可能存在相互作用。本研究通过检测两者在A549细胞迁移中的作用以及是否存在相互作用,为进一步研究它们在肿瘤发生发展中的作用机理提供实验依据... 背景与目的在前期研究中,我们发现在非小细胞肺癌中PRL-3和RhoC的表达具有相关性,提示两者可能存在相互作用。本研究通过检测两者在A549细胞迁移中的作用以及是否存在相互作用,为进一步研究它们在肿瘤发生发展中的作用机理提供实验依据。方法应用PRL-3抗体、RhoC抗体分别阻断A549细胞中PRL-3和RhoC的功能,采用细胞划痕实验检测两者对其迁移能力的影响;应用RT-PCR检测PRL-3和RhoC表达的变化。结果 PRL-3和RhoC功能被阻断后,A549细胞迁移能力明显降低;阻断PRL-3的A549细胞中RhoC表达降低,然而阻断RhoC的A549细胞中PRL-3表达无明显改变。结论 PRL-3和RhoC可能具有促进A549细胞远处转移的功能;PRL-3可能具有上调RhoC表达的功能;PRL-3促进癌细胞远处转移的功能可能是通过RhoC及其下游因子实现的,而RhoC对PRL-3的表达可能无反馈性调节作用。 展开更多
关键词 prl-3 细胞划痕实验 细胞培养
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蛋白酪氨酸磷酸酶PRL-3在乳腺癌中表达及意义 被引量:3
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作者 班振英 曾宪旭 +4 位作者 焦艳 党秋红 楚天骄 雷冬梅 王玉萍 《中华实用诊断与治疗杂志》 2012年第4期349-351,共3页
目的探讨乳腺浸润性导管癌及乳腺不典型增生组织中蛋白酪氨酸磷酸酶PRL-3表达情况。方法采用SP法及RT-PCR方法检测60份乳腺癌组织及50份乳腺不典型增生组织标本中PRL-3表达情况,分析PRL-3与乳腺浸润性导管癌临床病理特征的关系。结果在... 目的探讨乳腺浸润性导管癌及乳腺不典型增生组织中蛋白酪氨酸磷酸酶PRL-3表达情况。方法采用SP法及RT-PCR方法检测60份乳腺癌组织及50份乳腺不典型增生组织标本中PRL-3表达情况,分析PRL-3与乳腺浸润性导管癌临床病理特征的关系。结果在乳腺癌癌变过程中PRL-3表达阳性率呈递增趋势;PRL-3表达与有无淋巴结转移有相关性(P=0.011);PRL-3在乳腺癌中表达明显高于乳腺不典型增生组织(P<0.05)。结论乳腺癌组织中PRL-3在蛋白及基因水平呈高表达,一定程度上提示PRL-3与乳腺癌发生密切相关。 展开更多
关键词 乳腺癌 prl-3 RT-PCR
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