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Polyphyllin I inhibits proliferation and metastasis of ovarian cancer cell line HO-8910PM invitro

Journal of Traditional Chinese Medicine 2013年第3期33卷 325-333页

作者： Linhui Gu Jianguo Feng Haiyan Xu Min Luo Dan Su Cancer Research Institute Zhejiang Cancer Hospital

OBJECTIVE： To study the anticancer mechanism of polyphyllin I （PPI）, a Traditional Chinese Medicine, on the ovarian cancer cell line HO-8910PM in vitro. METHODS： Transwell chamber invasive assays were used to investigate the inhibitory capacity of PPI on HO-8910PM metastasis. Gene expression profiling chips was used to screen differentially ex- pressed genes between experiment group and con- trol group. Reverse transcription PCR and Western blotting were used to determine mRNA and pro- tein levels. RESULTS： With increasing PPI concentration, the metastatic capacity of cells decreased, with signifi- cance differences between the experimental and control groups （P〈0.01） as well as between two concentration groups. Gene expression profiling identified 123 differentially expressed genes, of which 70 were downregulated and 53 were upregu- lated. The genes were involved in multiple signal transduction pathways, including apoptosis, prolif- eration and metastasis. Real-time PCR （RT-PCR） showed that differential genes PIK3C2B, Caspase 9and WntSA were downregulated with increasing PPI, showing an evident dose-effect relationship. The c-Jun was an exception. As the PPI dosage in- creased and the exposure time was extended, c-Jun relative expression showed an upward trend. There were significant differences between the ex- periment and control （P〈0.05）. Western blot analy- ses showed that PPI treatment decreased levels of Caspase 9, WntSA and PIK3C2B and increased acti- vated Caspase 9,c-Jun and p-c-Jun expression levels. CONCLUSION＂ PPI has strong antitumor and anti transfer activity. It can activate c-Jun expression and the JNK signaling pathway, elicit cell apoptosis via the mitochondrial-mediated Caspase activation pathway, and finally inhibit tumor growth and mi- gration in vitro. The downregulation of PIK3C2B and Wnt5A jointly inhibit the proliferation and me- tastasis of HO-8910PM. PPI may be a novel treat- ment for ovarian cancer.

关键词： Chonglou Saponin I, Ovarian Neo-plasms Apoptosis cell movement

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Role of RhoA in platelet-derived growth factor-BB-induced migration of rat hepatic stellate cells

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Chinese Medical Journal 2010年第18期123卷 2502-2509页

作者： LI Lei LI Jing WANG Ji-yao YANG Chang-qing JIA Ming-lei JIANG Wei Department of Gastroenterology Zhongshan Hospital Fudan University Shanghai 200032 China Department of Gastroenterology Tongji Hospital Tongji University Shanghai 200065 China Department of Gastroenterology Huadong Hospital Fudan University Shanghai 200040 China

Background Although the migration of hepatic stellate cells （HSCs） is essential for hepatic fibrotic response, the detailed mechanisms involved are poorly understood. The aim of this study was to examine the role of Rho GTPases （especially RhoA） in platelet-derived growth factor （PDGF）-BB-induced migration of HSCs. Methods The migration of primary rat HSCs was evaluated using transwell Boyden chamber, while cytoskeletal changes were visualized by immunofluorescence staining of intracellular actins and vinculin. Quantitative real-time PCR and Western blotting analysis were used to detect the expression of Rho GTPases （RhoA, Racl and Cdc42） within HSCs and their activation was determined by glutathione S-transferase pull-down assay. Finally, the effects of RhoA on PDGF-BB-induced cell migration and cytoskeletal remodeling were analyzed using HSC-T6 cells stably transfected with constitutively active （CA, Q63L） or dominant negative （DN, T19N） RhoA mutants. Data were analyzed using SPSS 16.0 software. Student＇s t test was used to analyze differences between two groups and one-way analysis of variance （ANOVA） was used among multiple groups. Results Rapid cytoskeletal remodeling led to a significant increase in the motility of primary rat HSCs after haptotactic （direct） and chemotactic （indirect） stimulation by PDGF-BB： PDGF-BB caused a dramatic elevation in the levels of both total and active RhoA protein. However, the levels of mRNA for Rho GTPases, including RhoA, Racl and Cdc42, were unaffected. Furthermore, PDGF-BB induced increased formation of stress fibers and focal adhesions in HSC-T6 cells transfected with CA-RhoA, but not in HSC-T6 transfected with DN-RhoA. Surprisingly, both CA- and DN-RhoA-transfected HSC-T6 cells showed decreased migratory potential in the absence or presence of PDGF-BB compared with controls. Conclusions PDGF-BB induced cytoskeletal remodeling in rat HSCs and promoted their migration via regulation of intracellular RhoA. RhoA may be one of

关键词： platelet-derived growth factor BB hepatic stellate cells cell movement RhoA protein liver fibrosis

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Effects of aspirin on number,activity and inducible nitric oxide synthase of endothelial progenitor cells from peripheral blood

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Acta Pharmacologica Sinica 2006年第4期27卷 430-436页

作者： Tu-gang CHEN Jun-zhu CHEN~2 Xu-dong XIE Department of Cardiovascular Disease,First Affiliated Hospital,Medical School of Zhejiang University.Hangzhou 310003,China DepartmentofCardiovascularDisease FirstAffiliatedHospitalMedicalSchoolofZhejiangUniversityHangzhou310003China

Aim:To investigate whether aspirin has an influence on endothelial progenitorcells(EPC).Methods:Total mononuclear cells(MNC)were isolated from periph-eral blood by Ficoll density gradient centrifugation,then cells were plated onfibronectin-coated culture *** 7 d of culture,attached cells were stimu-lated with aspirin(to achieve final concentrations of 1,2,5,and 10 retool/L)for 3,6,12,and 24 *** were characterized as adherent cells that were double positivefor 1,1-dioctadecyl-3,3,3,3-tetramethylindocarbocyanine low density lipoprotein(DiLDL)uptake and lectin binding by direct fluorescent *** prolifera-tion and migration were assayed using a 3-(4,5-dimethyl-2 thiazoyl)-2,5-diphenyl-2H-tetrazolium bromide(MTT)assay and a modified Boyden chamber assay,*** EPC adhesion assay was performed by replating the EPC onfibronectin-coated dishes,and then adherent cells were *** vitrovasculogenesis activity was assayed by using an in vitro vasculogenesis *** nitric oxide synthase(iNOS)was assayed by Western ***:Incubation of isolated human MNC with aspirin decreased the number of *** also decreased the proliferative,migratory,adhesive,and in vitrovasculogenesis capacity of EPC,and also their iNOS levels in a concentration-and time-dependent ***:Aspirin decreases(1)the number ofEPC;(2)the proliferative,migratory,adhesive and in vitro vasculogenesis capaci-ties of EPC;and(3)iNOS levels in EPC.

关键词： aspirin endothelial progenitor cells cell adhesion cell count cell movement cell division physiologic neovascularization

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Effect of a nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester on invasion of human colorectal cancer cell line SL-174T

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World Journal of Gastroenterology 2005年第40期11卷 6385-6388页

作者： Li-Bo YU Xin-Shu Dong Wen-Zhou sun Dong-Lu Zhao Yue Yang Department of GeneralSurgery the Affiliated Tumor Hospital of Harbin MedicalUniversity Harbin 150040 Heilongiiang Province China Cancer Research Institute of HarbinMedical University Harbin 150040 Heilongjiang Province China

AIML To investigate the effect and mechanism of action of the nitric oxide synthase （NOS） inhibitor NG-nitro-L-arginine methyl ester （L-NAME） on invasion and metastasis of human colorectal cancer cell line SL-174T. METHODS： Human colorectal cancer cel4 line SL-174T was cultured and treated separately with four different dosages of L-NAME for 72 h, Nitric oxide （NO） production was measured with Griess reagent, The effect of L-NAME on invasion and migration of SL-174T cells were evaluated by using Transwell chambers attached with polycarbonate filters and reconstituted basement membrane （Matrigel）, RT-PCR was performed to determine the mRNA levels of matrix metalloproteinase-2 （MMP-2） and tissue inhibitor metalloproteinase-2 （TIMP-2）,RESULTS： L-NAME could significantly inhibit NO production of SL-174T in a dose-dependent manner. After being treated for 72 h with 0.2, 0.4, 0.8, and 1.0 mmol/L L- NAME, respectively, the ability of the L-NAME treated SL- 174T cells to invade the reconstituted basement membrane decreased significantly （t = 8.056, P〈0.05; t= 14.467, P〈0.01; t= 27.785, P〈0.01; and t= 29.405, P〈0.01, respectively） and the inhibition rates were 10.29%, 19.62%, 34.08%, and 42.23%, respectively. Moreover, L-NAME could inhibit migration of SL-174T cells, and the inhibition rates were 20.76%, 24.95%, 39.43%, and 46. 85% for L-NAME at 0.2, 0.4, 0.8, and 1.0 mmol/L, respectively （t = 15.116, P〈0.01）. In addition, after treatment with L-NAME, expression of MMP-2 mRNA was significantly decreased （t = 71.238, P〈0.01） and that of TIMP-2 mRNA was markedly increased （t = -13.020, P〈0.01）. CONCLUSION： L-NAME exerts anti-invasive and anti- metastatic effects on SL-174T cell line via downregulating MNP-2 mRNA expression and upregulating TIMP-2 mRNA expression.

关键词： Colorectal Neoplasms Neoplasm Invasiveness cell Line, Tumor cell movement Dose-Response Relationship, Drug Female Gelatinase A Humans Male NG-Nitroarginine Methyl Ester Nitric Oxide Nitric Oxide Synthase inhibitors Tissue Inhibitor of Metalloproteinase-2

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In vitro Effects of Nerve Growth Factor on Cardiac Fibroblasts Proliferation, cell Cycle, Migration, and Myofibroblast Transformation

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Chinese Medical Journal 2018年第7期131卷 813-817页

作者： Yong Zhao Chun-Hua Ding Department of Cardiology Zhongshan Hospital Affiliated with Guangzhou University of Chinese Medicine Zhongshan Guangdong 528400 China Department of Cardiology Aerospace Center Hospital Peking University Aerospace Clinical College of Medicine Beijing 100049 China

Background： Recent research indicates that nerve growth factor （NGF） promotes cardiac repair following myocardial infarction by promoting angiogenesis and cardiomyocyte survival. The purpose of this study was to investigate the effects of NGF on cardiac fibroblasts （CFs） proliferation, cell cycle, migration, and myofibroblast transformation in vitro. Methods： CFs were obtained from ventricles of neonatal Sprague-Dawley rats and incubated with various concentrations of NGF （0, 0.01,0.1, 1, 10, and 100 ng/ml; 0 ng/ml was designated as the control group）. cell proliferation and cell cycle of the CFs were measured by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide assay and flow cytometry （FCM）, respectively. A cell scratch wound model and transwell were carried out to observe effects of NGF on migration of CFs after 24 h of culture. Real-time polymerase chain reaction （RT-PCR） and Western blotting were used to measure a-smooth muscle actin （α-SMA） at mRNA and protein levels after CFs were incubated with various concentrations of NGF. Results： Expression of α-SMA measured by RT-PCR and Western blotting significantly increased in the 1 and 10 ng/ml NGF groups （P 〈 0.05）. Absorbance values of CFs showed that NGF did not influence the proliferation of CFs （The A490values were 0.178 ± 0.038, 0.182 ± 0.011,0.189 ± 0.005, 0. 178 ± 0.01 0, 0.185± 0.025, and 0.177 ± 0.033, respectively, in the 0, 0.01,0.1, 1, 10, and 100 ng/ml NGF groups [P = 0.800, 0.428, 0.981, 0.596, and 0.913, respectively, compared with control group]）, and FCM analysis showed that the percentage of CFs in G0/G1, S, and G2/M phases was not changed （P 〉 0.05）. The cell scratch wound model and transwell showed that CFs migration was not significantly different （P 〉 0.05）. Conclusion： NGF induces myofibroblast transformation but does not influence proliferation, cell cycle, or migration of CFs in vitro

关键词： cell Cycle cell movement cell Proliferation Fibroblasts Myofibroblasts Nerve Growth Factor

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Alterations in metastatic properties of hepatocellular carcinoma cell following H-ras oncogene transfection

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World Journal of Gastroenterology 2001年第3期7卷 335-339页

作者： Qing Wang~1 Zhi Ying Lin~2 Xiao Li Feng~3 ~1Department of Microbiology,Medical Center of Fudan University.the former Shanghai Medical University,Shanghai 200032,China ~2Liver Cancer Institute,Zhongshan Hospital,Shanghai 200032,China ~3Shanghai Institute of Biochemistry,Academy Sinica,Shanghai 200031,ChinaQing Wang earned master degree from Shanghai Medical University in 1996,now a senior lecturer of microbiology,specialized in the role of oncogcncs on tumor metastasis,having 8 papers published. Department of Microbiology Medical Center of Fudan University Shanghai 200032 China. zmswq@***

AIM: To demonstrate the relationship between H-ras oncogene and hepatocellular carcinoma (HCC) metastasis. METHODS: Activated H-ras oncogene was transfected into SMMC 7721, a cell line derived from human HCC, by calcium phosphate transfection method. Some metastasis-related parameters were detected in vitro, including adhesion assay, migration assay, expression of collagenase IV(c IV ase) and epidermal growth factor receptor (EGFR). RESULTS: The abilities of H-ras-transfected cell clones in adhesion to laminin (LN) or fibronectin (FN), migration, c IV ase secretion increased markedly, and the expression of EGFR elevated moderately. More importantly, these alterations were consistent positively with the expression of p21, the protein product of H-ras oncogene. CONCLUSION: H-ras oncogene could induce the metastatic phenotype of HCC cell in vitro to raise its metastatic potential.

关键词： Carcinoma, Hepatocellular cell Adhesion cell movement Gelatinase A Gelatinase B Gene Expression Regulation, Neoplastic Genes, ras Humans In Vitro Liver Neoplasms Phenotype Predictive Value of Tests Receptor, Epidermal Growth Factor Transfection

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Activated focal adhesion kinase involved in adhesion and migration of vascular smooth muscle cells stimulated by fibronectin

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Chinese Medical Journal 2002年第4期115卷 494-497页

作者：尹航汪丽蕙霍勇彭旭夏春芳唐朝枢北京大学第一医院心脏科北京100034

OBJECTIVE: To study the effects of focal adhesion kinase (FAK) phosphorylation on smooth muscle cells (SMCs) adhesion and migration stimulated by fibronectin. METHODS: Adhesion and migration of cultured SMCs were stimulated by different concentrations of fibronectin (FN), FAK and its phosphorylation were detected by immunoprecipitation and Western blot. FAK antisense oligodeoxynucleotides (ODNs) were transfected into SMCs by cationic lipid to investigate its modulatory effects on tyrosine phosphorylation. SMCs adhesion and migration were also measured by morphological enumeration and modified Boyden Chambers, respectively. RESULTS: FAK were expressed when SMCs adhesion and migration were successfully simulated by different concentrations of FN. FAK phosphorylation were detected only at 20 microg/ml FN or more. FAK antisense ODNs were transfected efficiently by cationic lipid and FAK phosphorylation was inhibited substantially. The SMCs migration rate in the 5 - 60 microg/ml FN groups was reduced by 17.89% - 27.67%. cell migration stimulated by FN at 10, 20, 40 and 60 microg/ml were reduced by 23.26%, 21.63%, 19.31% and 17.88%, respectively (P

关键词： Animals cell Adhesion cell movement cells, Cultured DNA, Antisense Dose-Response Relationship, Drug Enzyme Activation Fibronectins Focal Adhesion Kinase 1 Focal Adhesion Protein-Tyrosine Kinases Muscle, Smooth, Vascular Phosphorylation Protein-Tyrosine Kinase Rats Transfection

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Crk-associated substrate,vascular smooth muscle and hypertension

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Frontiers of Medicine 2008年第4期2卷 323-331页

作者： Dale D.TANG The Center for Cardiovascular Sciences Albany Medical CollegeNY 12208USA

Hypertension is characterized by vascular smooth muscle constriction and vascular remodeling involving cell migration,hypertrophy and ***-associated substrate(CAS),the first discovered member of the adapter protein CAS family,has been shown to be a critical cellular component that regulates various smooth muscle *** this review,the molecular structure and protein interactions of the CAS family members are *** for the role of CAS in the regu-lation of vascular smooth muscle contractility is *** stimulation induces CAS phosphor-ylation on Tyr-410 in arterial smooth muscle,creating the binding site for the Src homology(SH)2/SH3 protein CrkII,which activates neuronal Wiskott-Aldrich syn-drome protein(N-WASP)-mediated actin assembly and force *** functions of CAS in cell migra-tion,hypertrophy and growth are also *** tyrosine kinase(Abl),c-Src,focal adhesion kinase(FAK),proline-rich tyrosine kinase 2(PYK2),pro-tein tyrosine phosphatase-proline,glutamate,serine and threonine sequence protein(PTP-PEST)and SHP-2 have been documented to coordinate the phosphorylation and dephosphorylation of *** downstream signaling partners of CAS in the context of cell motility,hyper-trophy,survival and growth are also *** new findings establish the important role of CAS in the modulation of vascular smooth muscle ***,the upstream regulators of CAS may be new biologic targets for the development of more effective and specific treatment of cardiovascular diseases such as hypertension.

关键词： Crk-associated substrate protein cytoskeleton muscle contraction cell movement cardiovascular diseases

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Involvement of p38 mitogen-activated protein kinase in the regulation of platelet-derived growth factor -induced cell migration

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Frontiers of Medicine 2007年第3期1卷 248-252页

作者： GONG Xiaowei WEI Jie LI Yusheng CHENG Weiwei DENG Peng JIANG Yong Department of Pathophysiology Key Laboratory of Functional Proteomics of Guangdong ProvinceSouthern Medical UniversityGuangzhou 510515China

The aim of this study was to investigate the role of p38 mitogen-activated protein kinase(MAPK)in cell migration induced by platelet-derived growth factor(PDGF).Western blot was performed to detect the phosphorylation of p38 in NIH3T3 cells treated with PDGF.A Transwell cell migration system was used to determine the effects of PDGF treatment on the migration of NIH3T3 cells and the influence of p38 deficiency on this process in a p38 gene knockout(p38^(−/−))mouse embryonic fibroblast cell *** the stimulation of PDGF,the migration of NIH3T3 cells was significantly increased(Pcell migration was significantly blocked in p38 gene knockout(p38^(−/−))mouse embryonic fibroblasts(MEFs)(Pcells(p38+/+).p38 MAPK plays an important role in the regulation of cell migration induced by PDGF.

关键词： p38 mitogen-activated protein kinase plateletderived growth factor cell movement cytoskeleton signal transduction

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Experimental study on the adhesion, migration and three-dimensional growth of Schwann cells on absorbable biological materials

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Chinese Journal of Traumatology 2003年第4期6卷 209-212页

作者：王光林林卫杨志明裴福兴刘雷 Department of Orthopaedics the West China Hospital Sichuan UniversityChengdu 610041China Department of Obstetrics and Gynecology the West China HospitalSichuan UniversityChengdu 610041China Department of Orthopaedics the West China Hospital Sichuan UniversityChengdu 610041Chinabjective: To study the adhesion migration and three-dimentional growth of Schwann cells on PLA (polylactic acid) nonspinning fibre cloth and polyglycolic/ polylactic acid (PLGA) fibres.Methods: Schwann cells/ECM gel solution and PLA nonspinning fibre cloth and PLGA fibres pretreated by collagen polylysine and ECM were co-cultured. Then the migration and three-dimensional growth of Schwann cells on the fibres were observed under phase contrast microscope and laser scanning confocal microscope.Results: Schwann cell/ECM solution wascompounded with PLA nonspinning fibre cloth. With formation of gel most Schwann cells resided in the fibre net holes and adhered to the fibres to form a multiplayer-arranged Schwann cell column like Bungner band. Schwann cells could adhere to PLGA fibres and grew and migrated along the fibres. ECM gel could significantly increase the adhering and migrating cell number.Conclusions: ECM gel can facilitate the adhesion growth and migration of Schwann cells on the seteroframe. It is a good integrating material for tissue engineering bioartificial nerve.

OBJECTIVE: To study the adhesion, migration and three-dimentional growth of Schwann cells on PLA (polylactic acid) nonspinning fibre cloth and polyglycolic/polylactic acid (PLGA) fibres. METHODS: Schwann cells/ECM gel solution and PLA nonspinning fibre cloth and PLGA fibres pretreated by collagen, polylysine and ECM were co-cultured. Then the migration and three-dimensional growth of Schwann cells on the fibres were observed under phase contrast microscope and laser scanning confocal microscope. RESULTS: Schwann cell/ECM solution was compounded with PLA nonspinning fibre cloth. With formation of gel, most Schwann cells resided in the fibre net holes, and adhered to the fibres to form a multiplayer-arranged Schwann cell column like Bungner band. Schwann cells could adhere to PLGA fibres and grew and migrated along the fibres. ECM gel could significantly increase the adhering and migrating cell number. CONCLUSIONS: ECM gel can facilitate the adhesion, growth and migration of Schwann cells on the seteroframe. It is a good integrating material for tissue engineering bioartificial nerve.

关键词： Tissue Engineering cell Adhesion cell Division cell movement Humans Lactic Acid Polyglycolic Acid Polymers Research Support, Non-U.S. Gov't Schwann cells

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