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Expression of thymidine kinase mediated by a novel non-viral delivery system under the control of vascular endothelial growth factor receptor 2 promoter selectively kills human umbilical vein endothelial cells

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World Journal of Gastroenterology 2008年第2期14卷 224-230页

作者： Ying Wang Hui-Xiong Xu Ming-De Lu Qing Tang Department of Medical Ultrasonics The First Affiliated Hospital Sun Yat-Sen UniversityGuangzhou 510080 Guangdong Province China Department of Hepatobiliary Surgery The First Affiliated Hospital Sun Yat-Sen University Guangzhou 510080Guangdong Province China

AIM: To investigate the killing efficiency of a recombinant plasmid containing a thymidine kinase (TK) domain insert driven by the vascular endothelial growth factor receptor 2 (VEGFR2) promoter (KDR) on vascular endothelial ***: The KDR-TK fragment was extracted from pBluescript Ⅱ KDR-TK plasmid by enzymatic digestion with Xho I and Sal I. The enhanced green fluorescence protein (EGFP) carrier was extracted from pEGFP by the same procedure. The KDR-TK was inserted into the pEGFP carrier to construct pEGFP-KDR-TK. Using ultrasound irradiation and microbubble, pEGFP-KDR-TK was transferred into human umbilical vein endothelial cells (HUVECs). The transient infection rate was estimated by green fluorescent protein (GFP) expression. Transfected HUVECs, non-transfected HUVECs, and HepG2 cells were cultured in the presence of different concentrations of ganciclovir (GCV), and the killing efficacy of HSV-TK/GCV was analyzed by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The recombinant pEGFP-KDR-TK was successfully constructed by inserting the KDR-TK fragment into the pEGFP carrier. Transfected HUVECs showed cytoplasmic green fluorescence, and the transient transfection rate was about 20.3%. Pools of G418-resistant cells exhibited a higher sensitivity to theprodrug/GCV compared to non-transfected HUVECs or non-transfected HepG2 cells, respectively. CONCLUSION: KDR promoter and the suicide gene/prodrug system mediated by diagnostic ultrasound combined with microbubble can significantly kill HUVECs. Such therapy may present a novel and attractive approach to target gene therapy on tumor vessels.

关键词： Microbubble Ultrasound Gene therapy vascular endothelial growth factor receptor 2 Humanumbilical vein endothelial cells

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Discovery of antitumor diterpenoids from Casearia graveolens targeting VEGFR-2 to inhibit angiogenesis

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Chinese Journal of Natural Medicines 2024年第9期22卷 842-853页

作者： WANG Sibei LIU Yuhui LIANG Yue XI Yaru ZHAI Yupeng LEE Dongho XU Jing GUO Yuanqiang State Key Laboratory of Medicinal Chemical Biology College of Pharmacyand Tianjin Key Laboratory of Molecular Drug ResearchNankai UniversityTianjin 300350China State Key Laboratory of Functions and Applications of Medicinal Plants Guizhou Medical UniversityGuiyang 550014China Department of Plant Biotechnology College of Life Sciences and BiotechnologyKorea UniversitySeoul 02841South Korea

Eight novel clerodane diterpenoids(1-8)were isolated from the twigs of Casearia *** structures were elucidated through comprehensive nuclear magnetic resonance(NMR),high-resolution electrospray ionization mass spectrometry(HRESI-MS),and electronic circular dichroism(ECD)*** addition to structural determination,surface plasmon resonance(SPR)assays were conducted to investigate molecular interactions,revealing that compound 8 exhibited high affinity for vascular endothelial growth factor receptor 2(VEGFR2),a key regulator of tumor *** in vivo experiments demonstrated that compound 8 effectively inhibited angiogenesis and displayed significant antitumor activity by suppressing tumor proliferation and metastasis in zebrafish xenograft *** findings suggest that compound 8 holds promise as an anticancer lead compound targeting VEGFR-2 to obstruct tumor angiogenesis.

关键词： Clerodane diterpenoids Casearia graveolens Surface plasmon resonance vascular endothelial growth factor receptor 2 Antitumor activity Zebrafish

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Targeted agents for second-line treatment of advanced hepatocellular carcinoma

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World Journal of Gastrointestinal Oncology 2019年第10期11卷 788-803页

作者： Nicola Personeni Tiziana Pressiani Silvia Bozzarelli Lorenza Rimassa Medical Oncology and Hematology Unit Humanitas Cancer CenterHumanitas Clinical and Research CenterIRCCSRozzano 20089MilanItaly Department of Biomedical Sciences Humanitas UniversityPieve Emanuele 20090MilanItaly

Over the past ten years,sorafenib,a multikinase inhibitor,has been the standard of care for patients with unresectable hepatocellular carcinoma(HCC)and wellpreserved liver ***,lenvatinib,a different multikinase inhibitor,was shown to be non-inferior to sorafenib,in terms of survival,while all other agents previously tested failed to prove non-inferiority(or superiority)when compared to ***,in the second-line setting,most investigational drugs failed to provide better survival outcomes than ***,in the last 2 years three positive phase III trials have been published in this *** RESORCE trial,a phase III study evaluating regorafenib in HCC patients who experienced disease progression after first-line treatment with sorafenib,showed better outcomes with regorafenib compared to *** recently,the phase III CELESTIAL trial demonstrated the superiority of cabozantinib,a multikinase inhibitor targeting vascular endothelial growth factor receptor,MET,and AXL,vs placebo in the second-and third-line setting in patients progressing on or intolerant to *** survival benefits of a sustained anti-angiogenic inhibition were demonstrated also with ramucirumab in the phase III REACH-2 trial in patients previously treated with sorafenib and who had high baseline alpha-fetoprotein ***,the adverse events reported in these trials were in line with the known safety profiles of the tested *** nearly a decade of a certain degree of stagnation,we are now witnessing a period of novel therapeutic advances with multikinase inhibitors and monoclonal antibodies that will likely change the treatment scenario of HCC.

关键词： Hepatocellular carcinoma Advanced-metastatic Second-line Third-line Regorafenib Cabozantinib Ramucirumab Angiogenesis Multikinase inhibitor MET AXL vascular endothelial growth factor receptor 2

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Intravitreal injection of resveratrol inhibits laser-induced murine choroidal neovascularization

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International Journal of Ophthalmology(English edition) 2020年第6期13卷 886-892页

作者： Hui-Ming Zhang Xiao-Hua Li Mingjiazi Chen Jing Luo Department of Dermatology the Second Xiangya HospitalCentral South UniversityChangsha 410011Hunan ProvinceChina Hunan Key Laboratory of Medical Epigenomics Changsha 410011Hunan ProvinceChina Department of Pathology and Ophthalmology Keck School of Medicine at University of Southern CaliforniaLos AngelesCalifornia 90033USA Henan Provincial People's Hospital/Henan Eye Hospital Henan Eye InstituteHenan Key Laboratory of Ophthalmology and Vision Science/Department of OphthalmologyPeople’s Hospital ofZhcngzliou University/Department of OphthalmologyPeople's Hospital of Henan UniversityZhengzhou 450003Henan ProvinceChina 5Department of Ophthalmologythe Second Xiangya I IospitalCentral South UniversityChangsha 410011Hunan ProvinceChina

AIM:To determine the effects of intravitreal resveratrol(RSV)on murine laser-induced choroidal neovascularization(CNV).METHODS:The toxicity of RSV to choroidal endothelial cell(CEC)was measured using thiazolyl blue tetrazolium bromide(M一)*** of RSV on choroidal endothelial cell(CEC)migration were evaluated with a modified Boyden chamber assay,while tube formation was evaluated in a 2-D gel *** was induced by laser photocoagulation in *** effects of intravitreal injection of RSV on CNV development were evaluated by fluorescein angiography(FA),confocal analysis of isolectin B4 labeled choroidal flat mounts,and histologic examination of CNV *** was used to analyze the expression and phosphorylation of vascular endothelial growth factor receptor 2(VEGFR2).RESULTS:No significant cell toxicity was observed in CEC if the concentration of RSV was less than 200 pmol/L(P>0.05).RSV inhibited vascular endothelial growth factor(VEGF)-induced CEC migration(P2,178163±78623μm^3 and 6508±619.0μm^2,respectively)compared with those in control(2.724±0.08447,379676±98382μm3and16576±2646μm^2,respectively;P2expression was much weaker in the sections of CNV lesions in RSV injected mice compared with that in control(Pendothelial cell migration,tube formation and VEGFR2 phosphorylation.

关键词： resveratrol intravitreal injection choroidal neovascularization vascular endothelial growth factor receptor 2 tube formation murine

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Apatinib as an alternative therapy for advanced hepatocellular carcinoma

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World Journal of Hepatology 2020年第10期12卷 766-774页

作者： Xi-Hao Zhang Man-Qing Cao Xiu-Xiu Li Ti Zhang Tianjin Medical University Cancer Institute and Hospital National Clinical Research Center for CancerTianjin’s Clinical Research Center for CancerTianjin 300060China Key Laboratory of Breast Cancer Prevention and Therapy Tianjin Medical UniversityTianjin 300060China Department of Breast Surgery Tianjin Medical University Cancer Institute and HospitalNational Clinical Research Center for CancerTianjin’s Clinical Research Center for CancerTianjin 300060China Digestive Department Shanxi Province Tumor HospitalTaiyuan 030013Shanxi ProvinceChina Department of Hepatobiliary Surgery Tianjin Medical University Cancer Institute and HospitalTianjin 300060China

Angiogenesis plays an important role in the occurrence and development of *** tyrosine kinase inhibitors targeting vascular endothelial growth factor reduce ***,a tyrosine kinase inhibitor,can specifically inhibit vascular endothelial growth factor receptor 2,showing encouraging anti-tumor effects in a variety of tumors including advanced hepatocellular carcinoma(HCC).This article intends to review the clinical research and application prospects of apatinib in the field of HCC.

关键词： Apatinib Hepatocellular carcinoma Angiogenesis vascular endothelial growth factor receptor 2

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Effect of VEGF on Neural Differentiation of Human Embryonic Stem Cells in vitro

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Journal of Huazhong University of Science and Technology(Medical Sciences) 2009年第5期29卷 563-566页

作者：焦淑洁许慧芳许杰湛彦强张苏明 Department of Neurology Tongji HospitalTongji Medical CollegeHuazhong University of Science and Technology

The effects of vascular endothelial growth factor （VEGF） on neural differentiation of human embryonic stem cells （hESCs） in vitro and the possible mechanism were observed. The hESCs lines, TJMU1 and TJMU2, were established and stored by our laboratory, hESCs differentiated into neuronal cells through embryonic body formation. In this induction process, hESCs were divided into three groups： group A, routine induction; group B, routine induction＋10 ng/mL VEGF; group C, routine in- duction＋10 ng/mL VEGF＋10 ng/mL VEGFR2/Fc. OCT4, Nestin and GFAP in each group were de- tected by RT-PCR, and the cells expressing Nestin and GFAP were counted by immunofluorescence. The percentage of Nestin positive cells in group B was significantly higher than in groups A and C, while the percentage of GFAP positive cells in group B was significantly lower than in groups A and C （P〈0.01）. There was no significant difference between groups A and C （P〉0.05）. It was concluded that VEGF, via VEGFR2, stimulated the neural differentiation of hESCs in vitro.

关键词： human embryonic stern cell differentiation vascular endothelial growth factor vascular endothelial growth factor receptor 2

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Combination therapy with VEGFR2 and EGFR siRNA enhanced the antitumor effect of cisplatin in non-small cell lung cancer xenografts

Combination therapy with VEGFR2 and EGFR siRNA enhanced the ...

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2014年广东省药师周大会

作者： Shan Chen Xiaoxia Liu Wenfei Gong Huan Yang Daofei Luo Xialin Zuo Wenshi Li Ping Wu Li Liu Qian Xu Aimin Ji Department of Pharmaceutical Zhujiang HospitalSouthern Medical University

siRNA targeting the murine VEGFR2 gene was designed and validated for efficient and robust silencing in vitro,and was then delivered by PEI in vivo to investigate the anti-tumor effect on non-small-cell lung cancer *** following dosage regimens were tested for their tumor inhibitory effect in vivo:VEGFR2 siRNA,EGFR siRNA,VEGFR2 siRNA + EGFR siRNA,cisplatin alone,and VEGFR2 siRNA + EGFR siRNA + *** silencing of both VEGFR2 and EGFR expression by siRNA,combined with low dose cisplatin was found to effectively inhibit tumor growth,and extend the survival time of mice bearing the NSCLC *** results suggest that combination therapy using siRNAs and chemotherapy agents might offer a novel strategy for cancer treatment in the future.

关键词： siRNA vascular endothelial growth factor receptor 2 epidermal growth factor receptor chemotherapy agents

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Sprouty4 regulated vascular permeability by regulating the tyrosine phosphorylation and degradation of VE-cadherin

Sprouty4 regulated vascular permeability by regulating the t...

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中国生物化学与分子生物学会2019年全国学术会议暨学会成立四十周年

作者： Jiangbo Ren Nannan Zhang Wenjie Sun Yuan Luo Yuke Gao Yan Gong Department of Biological Repositories Zhongnan Hospital of Wuhan University Department of Radiation and Medical Oncology Zhongnan Hospital of Wuhan University

Regulation of endothelial cell adherent junctions is important to pathophysiological processes, in which vascular integrity is disrupted such as during tumor angiogenesis and chronic inflammatory conditions. The effects of Sprouty4(Spry4) on endothelial barrier function was investigated using vascular endothelial cell growth factor(VEGF)-induced vascular permeability model. In the present study we showed that overexpression of Spry4 inhibited vascular permeability in primary human endothelial cells in vitro and in Spry4-expressing transgenic mice in vivo. Conversely, knockdown of Spry4 in vitro and conditional endothelial cell-specific knockout of Spry4 in endothelial cells in vivo enhanced VEGF-induced vascular permeability. In addition, overexpression of Spry4 inhibited VEGF-induced vascular endothelial cadherin(VE-cadherin) tyrosine phosphorylation, in part by inhibiting c-Src activation, and resistance to VEGF-induced vascular permeability. Inhibition of VEGF-induced VE-cadherin tyrosine phosphorylation and decreased vascular permeability by Spry4 overexpression could be rescued by expression of constitutively active c-Src. Conversely, knockdown of Spry4 in endothelial cells increased tyrosine phosphorylation of VE-cadherin and vascular permeability, which was repressed by dominant negative Src(DNSrc). Consistent with these results, DNSrc repressed the increase in vascular permeability that resulted due to knockdown of Spry4. Taken together, these results suggested that Spry4 regulated vascular permeability via regulating the activation of c-Src by VEGF signaling and consequently the tyrosine phosphorylation of VE-cadherin.

关键词： vascular integrity endothelial permeability vascular endothelial growth factor receptor 2 vascular endothelial cadherin Sprouty

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