RGD对人纤维肉瘤细胞HT1080增殖、黏附及转移影响的研究

利用合成的RGD(Arg-Gly-Asp)三肽研究了其抑制人纤维肉瘤细胞HT1080的增殖、黏附及转移作用.采用MTT法检测了不同浓度的RGD对HT1080细胞增殖及黏附纤维粘连蛋白(fibronectin)能力的影响;采用细胞划痕法研究了RGD对HT1080细胞在纤维粘连蛋白中迁移能力的影响.结果表明,合成的RGD能抑制HT1080细胞的增殖,并具有抗肿瘤细胞黏附、抗转移的活性.     

穗花杉双黄酮抑制人脐静脉内皮细胞血管形成的实验研究

为探讨穗花杉双黄酮(Amentoflame,AF)对人脐静脉内皮细胞ECV304血管形成及机制的影响,从而为穗花杉双黄酮治疗肿瘤及其他血管增生性疾病提供理论基础,采用MTS检测AF对ECV304血管内皮细胞增殖的作用,细胞划痕实验观察AF对ECV304细胞迁移的影响,体外血管形成实验观察AF对ECV304血管形成的影响,Western blot检测AF对ECV304细胞血管形成相关信号通路及蛋白如磷酸化AKT(p-AKT)、金属基质蛋白酶9(Matrix metalloproteinase-9,MMP-9)、促血管生成素2(angiopoietin 2,Ang 2)表达影响。结果表明AF能够抑制ECV304细胞的增殖作用,并呈梯度依赖性;AF具有抑制ECV304细胞迁移的作用,并且能够抑制ECV304体外血管样结构的形成;Western blot结果显示100μmol/L的AF能够降低ECV304细胞p-AKT、MMP-9、Ang-2的蛋白表达。     

血管生成抑制素对体外培养的血管内皮细胞增殖与凋亡的影响

为研究血管生成抑制素对体外培养的血管内皮细胞生长的影响,采用MTT法观察细胞的增殖情况,利用Hoest染色和流式细胞仪检测细胞凋亡。结果发现血管生成抑制素能够抑制血管内皮细胞的增殖,其IC50为1.19 mg/L,并干扰内皮细胞的周期,出现G0/G1期阻滞。     

福建大头蛙抗癌多肽FJ-2945抗肿瘤活性研究

探索本课题组自主发现的天然抗肿瘤多肽FJ-2945对肝癌细胞Hep3b增殖、迁移的影响并初步明确其作用机制.通过CCK8与RTCA实验检测FJ-2945对细胞增殖能力的影响;采用Transwell与划痕实验分析FJ-2945对于Hep3b迁移能力的影响;利用流式细胞分析法检测FJ-2945对Hep3b细胞周期的影响;最后通过qRT-PCR与免疫印迹实验检测FJ-2945对Hep3b细胞增殖、迁移相关因子在转录水平与蛋白水平的影响.与对照组相比,FJ-2945显著抑制Hep3b细胞的增殖与迁移能力,并促进Hep3b细胞凋亡.实验组中的Skp2蛋白含量显著降低,最终抑制Hep3b细胞增殖与迁移能力.由此说明多肽FJ-2945可以作为潜在的药物,通过靶向Skp2抑制肝癌细胞Hep3b增殖、迁移等过程,发挥抗肿瘤作用.     

Napabucasin对结直肠癌细胞增殖及迁移的影响

为了探究新型小分子抑制剂Napabucasin对结直肠癌细胞增殖以及迁移的影响. 首先通过分子模拟对接分析了Napabucasin与STAT3蛋白的互作机制. 然后利用克隆形成实验、细胞划痕实验等方法在多种结直肠癌细胞系中证明了Napabucasin能够显著抑制结直肠癌细胞的集落形成能力以及迁移能力. 进而使用Napabucasin与Wnt信号通路激活剂Wnt agonist 1共处理结直肠癌细胞HCT116,结合蛋白质印迹实验发现,Wnt信号通路介导了Napabucasin对结直肠癌细胞的迁移以及增殖的抑制过程. 研究结果显示,Napabucasin能够在体外抑制结直肠癌细胞的增殖能力以及迁移能力,并且Wnt信号通路参与介导了这一抑制过程.     

高迁移率族蛋白1对脐静脉内皮细胞迁移及相关蛋白表达的影响

【目的】组织工程中微循环的建立在生物材料植入中发挥重要作用，其中血管内皮细胞迁移是快速血管生成的主要影响因素。组织和器官损伤后常引发炎症反应，高迁移率族蛋白1(HMGB1)是炎症反应的起始因子，但对内皮细胞迁移的影响尚不清楚。【方法】体外提取和培养人脐静脉内皮细胞(HUVECs),并进行细胞表型鉴定；构建培养液中含有HMGB1的浓度梯度，通过CCK-8、划痕实验和Transwell小室实验检测HUVECs的增殖能力和迁移能力，采用RT-qPCR和Western Blot检测细胞内细胞增殖因子和迁移因子基因和蛋白的表达情况。【结果】不同浓度HMGB1对HUVECs的增殖能力无显著影响，但对细胞迁移能力有浓度依赖性，随着HMGB1浓度增加，细胞的迁移能力增强。FGF-2表达随HMGB1浓度增加表达上调；高浓度HMGB1处理HUVECs后，VEGFA表达明显下降。【结论】HMGB1可通过FGF-2促进HUVECs迁移，为今后HMGB1对内皮细胞迁移能力的影响研究提供依据。     

桑叶对乳腺癌肿瘤血管内皮细胞抑制作用的研究

本文通过流式细胞仪(FCM)和酶联免疫吸附法(ELISA)来研究桑叶对乳腺癌肿瘤血管内皮细胞(ECs)细胞凋亡、细胞周期及内皮细胞生长因子受体-2(VEGFR-2)的影响.FCM分析表明,桑叶中有效成分能够诱导异常增殖的内皮细胞凋亡,使细胞周期停滞在DNA合成期,有效阻止内皮细胞的有丝分裂(P0.05);ELISA法结果表明,VEGFR-2的表达受到明显的抑制(P0.05).以上结果证实桑叶能够有效促进乳腺癌肿瘤血管内皮细胞的凋亡、影响其细胞周期的分布、抑制乳腺癌肿瘤血管内皮细胞生长因子受体-2的表达,对其增殖具有抑制作用.     

茶溴香酰胺脂质体对黑色素瘤细胞生长和迁移的抑制作用

考察茶溴香酰胺脂质体(TBrC-L)对黑色素瘤B16细胞生长和迁移的抑制作用及其分子机制,为研发抗癌新药提供科学依据.采用MTT法和细胞划痕术探究TBrC-L对黑色素瘤B16细胞生长和迁移的影响; Hoechst 33258染色法观察药物作用下细胞凋亡形态的变化; Western Blotting检测TBrC-L对B16细胞生长、迁移和凋亡相关蛋白表达的影响.结果表明:TBrC-L能对B16细胞的生长和迁移产生显著性抑制作用,并且诱导其凋亡; TBrC-L可下调c-Met、HGF、Bcl-2、NF-κB和VEGFR2的表达,上调Caspase-3、E-cadherin、Bax、p53和Cytochrome c的表达,抑制HGF/Met/VEGFR2-NF-κB通路可能是TBrC-L抑制黑色素瘤B16细胞生长和迁移作用的重要分子机制.本研究结果提示,TBrC-L具有进一步开发为抗黑色素瘤药物的潜力.     

三维自组装石墨烯的细胞相容性研究

针对石墨烯的二维平面结构不利于细胞在材料上延伸生长和黏附的问题,通过水热法合成自组装石墨烯,搭建利于细胞生长和黏附的三维结构.利用扫描电子显微镜、透射电子显微镜、傅里叶红外光谱、拉曼光谱、X线光电子能谱和接触角测试仪对样品的表面形貌、微观结构、化学键、原子组态和亲水性能进行测试,并通过形体外细胞实验研究了L929小鼠肺部成纤维细胞在材料上的黏附和增殖生长等行为.实验结果表明:自组装石墨烯具有孔径为1~3μm的孔洞,材料整体呈网状结构,为多层石墨烯片层的堆叠;样品中部分C元素分别与H和O元素结合生成多种官能团;三维自组装石墨烯的亲水性和表面细胞增殖生长情况均优于二维石墨烯,更适于作为组织支架促进细胞生长.     

电刺激对内皮细胞及血管组织的作用

电学环境是生物体所处的重要微环境之一,外加电刺激对内皮细胞和血管有重要作用.电刺激对细胞活性的影响已越来越引起人们的兴趣和重视,同时在生物科学中已得到了广泛的应用.文中就电刺激对内皮细胞粘附、增殖、迁移和分泌物产生的影响以及对血管收缩和舒张的影响及其机理作些介绍和分析.     

Effects of Genistein and Daidzein on the Proliferation, Invasion, Migration and Adhesion of Melanoma Cells

IntroductionGenistein and daidzein (Fig.1 ) ,two majorisoflavonoids in soybeans,were reported to playimportant roles in cancer prevention[1 ,2 ] .Previousstudies demonstrated that genistein and daidzeininhibited the growth of leukemia,breast,colonand prostate cancers[38] .Our earlier studies alsodemonstrated that daidzein enhanced the immunefunction in mice[9,1 0 ] . These functions madegenistein and daidzein promising candidates forcancer prevention. However,it is usually themetastatic disse…     

Genistein inhibits human TNF-α-induced porcine endothelial cell adhesiveness for human monocytes and natural killer cells

Cellular immune response is a major barrier to xenotransplantation. Human tumor necrosis factor-α (hTNF-α) possesses cross-species activity and directly amplifies the immune rejection via the upregulation of adhesion molecules on porcine endothelium. We investigated the role of protein tyrosine phosphorylation in the induction of expression of E-sclectin and vascular cell adhesion molecule-1 (VCAM-1), and the augmentation of adhesion of human peripheral blood monocytes (PBMo) and natural killer cells (PBNK), after rhTNF-α-stimulation of porcine aortic endothelial cells (PAEC) in vitro, rhTNF-α-increased adhesiveness of PAEC for both PBMo and PBNK was dose-dependently reduced by pretreatment of PAEC with the selective protein tyrosine kinase (PTK) inhibitor genistein. The inhibitory effect occurred at the early time of PAEC activation triggered by rhTNF-α, and was completely reversible. PTK activity assay indicated that genistein also suppressed rhTNF-α stimulated activation of protein tyrosine kinases (PTKs) in PAEC in a dose-dependent manner. Flow cytometric analysis showed that genistein inhibited the upregulation of E-selectin and VCAM-1 by rhTNF-α. These results suggest that PTKs may regulate the expression of E-selectin and VCAM-1 on PAEC and the adherence of PBMo and PBNK induced by rhTNF-α. Moreover, dietary genistein, used as an adhesion antagonist, may contribute to managing the cell-mediated rejection in the clinical application.     

RhoGTPases in stem cells

RhoGTPases are small molecules that control a wide variety of signal transduction pathways. Their profound function in regulating the actin cytoskeleton is well recognized. Stem cells are unique in their ability to self-renew and produce progenitor cells that can differentiate into specialized cells. RhoGTPases influence stem cell morphology and cell migration as well as stem cell self-renewal, proliferation, transplantation, homing and differentiation. In this review, the multiple roles of the RhoGTPases in stem cells are discussed.     

木棉皮抗消化道肿瘤活性部位筛选及抑制肿瘤转移机制

为筛选木棉皮醇提物抗消化道肿瘤活性部位,探究其抑制敏感肿瘤细胞增殖、转移作用机制,通过采用CCK-8(cell counting kit-8)法考查木棉皮醇提物不同极性萃取部位对人肝癌HepG2细胞、人胃癌SGC7901细胞、人结肠癌SW480细胞和人胰腺癌PANC-1细胞这4种肿瘤细胞的抑制作用,筛选出木棉皮醇提物抗肿瘤的活性部位和敏感细胞。采用细胞划痕实验、Transwell实验和细胞黏附实验,研究木棉皮醇提物抗肿瘤活性部位对敏感肿瘤细胞迁移、侵袭和黏附能力的影响。定量聚合酶链反应法(quantitative polymerase chain reaction, qPCR)检测基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)基因的mRNA转录水平。结果表明,木棉皮醇提物不同极性萃取部位中石油醚部位对人胃癌SGC7901细胞和人肝癌HepG2细胞的抑制率最大,且对人胃癌SGC7901细胞最为敏感。随着木棉皮醇提物石油醚部位浓度增加,在24、48、72 h时间段人胃癌SGC7901细胞的迁移面积相对于空白组有所减小(P<0.05)。与空白组相比,木棉皮醇提物石油...     

Effects of Genistein on Cell Cycle and Apoptosis of Two Murine Melanoma Cell Lines

The effects of genistein on several tumor cell lines were investigated to study the effects of gen- istein on cell growth, cell cycle, and apoptosis of two murine melanoma cell lines, B16 and K1735M2. These two closely related murine melanoma cell lines, however, have different responses to the genistein treat- ment. Genistein inhibits the growth of both the B16 and K1735M2 cell lines and arrests the growth at the G2/M phase. After treatment with 60 μmol/L genistein for 72 h, apoptosis and caspase activities were de- tected in B16 cells, while such effects were not found in K1735M2. Further tests showed that after genistein treatment the protein content and mRNA levels of p53 increased in B16, but remained the same in K1735M2. The protein content and mRNA levels of p21WAF1/CIP1 increased in both cell lines after treatment. The results show that genistein might induce apoptosis in B16 cells by damaging the DNA, inhibiting topoi- somerase II, increasing p53 expression, releasing cytochrome c from the mitochondria, and activating the caspases which will lead to apoptosis.     

Multiple Suppressive Effects of a Protein from Caesalpinia minax on Murine Melanoma Cells

IntroductionIn recent years,a great deal of attention hasfocused on finding potential anti- tumor agentsfrom natural sources[14 ] .A vast number ofpromising candidate molecules,especiallycomponents extracted from plants,have beenevaluated[59] .　 Herbs have a long history of use as Chinesetraditional medicine.Caesalpinia minax (C.minax) is a wild plantin Yunnan Province,China.The high level of ultraviolet radiation in thistropical province causes widespread skin diseases inthis area.Extract…     

Roles for microtubule and microfilament cytoskeletons in animal cell cytokinesis

Microtubule and microfilament cytoskeletons play key roles in the whole process of cytokinesis. Although a number of hypotheses have been proposed to elucidate the mechanism of cytokinesis by microtubule and actin flament cytoskeletons, many reports are conflicting. In our study,combining the cytoskeletons drug treatments with the time-lapse video technology, we retested the key roles of microtubule and actin filament in cytokinesis. The results showed that depolymerization of microtubules by Nocodazole after the initiation of furrowing would not inhibit the furrow ingression, but obviously decrease the stiffness of daughter cells. Depolymerizing actin filaments by Cytochalasin B before metaphase would inhibit the initiation of furrowing but not chromosome segregation, resulting in the formation of binucleate cells; however, depolymerizing actin fillaments during anaphase would prevent furrowing and lead to the regress of established furrow, also resulting in the formation of binucleate cells. Further, depolymerizing microtubules and actin filaments simultaneously after metaphase would cause the quick regress of the furrow and the formation of binudeate cells. From these results we propose that a successful cytokinesis requires functions and coordination of both the microtubule and actin filament cytoskeletons.Microtubule cytoskeleton may function in the positioning and initiation of cleavage furrow, and the actin filament cytoskeleton may play key roles in the initiation and ingression of the furrow.     

Interaction of plasma membrane fibronectin receptor with talin--a transmembrane linkage

Many observations suggest the presence of transmembrane linkages between the cytoskeleton and the extracellular matrix. In fibroblasts both light and electron microscopic observations reveal a co-alignment between actin filaments at the cell surface and extracellular fibronectin. These associations are seen at sites of cell matrix interaction, frequently along stress fibres and sometimes where these bundles of microfilaments terminate at adhesion plaques (focal contacts). Non-morphological evidence also indicates a functional linkage between the cytoskeleton and extracellular matrix. Addition of fibronectin to transformed cells induces flattening of the cells and a reorganization of the actin cytoskeleton, with the concomitant appearance of arrays of stress fibres. Conversely, disruption of the actin cytoskeleton by treatment with cytochalasin B leads to release of fibronectin from the cell surface. As yet, there is no detailed knowledge of the molecules involved in this transmembrane linkage, although several proteins have been suggested as candidates in the chain of attachment between bundles of actin filaments and the cytoplasmic face of the plasma membrane: these include vinculin, alpha-actinin and talin, each one having been identified at regions where bundles of actin filaments interact with the plasma membrane and underlying cell-surface fibronectin. Recently, the cell-substrate attachment (CSAT) antigen has been identified as a plasma membrane receptor for fibronectin, raising the possibility that this glycoprotein complex may serve as a bridge between fibronectin and one or more of the underlying cytoskeletal components mentioned. Here we have investigated the interaction of the purified CSAT antigen with these cytoskeletal components, and we demonstrate an interaction specifically between the CSAT antigen and talin.     

Cytoskeleton reorganization and FAK phosphorylation are involved in lanthanum(Ⅲ)-promoted proliferation and differentiation in rat osteoblasts

The effect of lanthanum ion(La3+)on osteoblast function and cytoskeleton is assessed in vitro. Osteoblasts were isolated from Snraeuo-tjawiey tetai neonatal rats.Cell proliferation and gene expression levels of cbfa-1,alkaline phosphatase(ALP),osteocalcin(OC),bone sialo-protein(BSP)and osteopontin(OPN)were examined by cell counting and RT-PCR.Cytoskeleton F-actin was stained with rhodamine-con-jugated phalloidin and was visualized by a confocal microscope.As the results,10-8-10-4M La3+-induced osteoblast proliferation on day2.Data from the RT-PCR assay revealed that 10-6 M La3+ up-regulated the expression levels of ALP,BSP,and cbfa-lon day 4,while it enhanced the expressions of OC and OPN on day 21.The F-actin cytoskeleton was strengthened and reorganized under the exposure of La3+.In addition,the phosphorylation of focal adhesion kinase(FAK)was significantly promoted in 24 h evaluated by Western blot analysis.These findings indi-sate that La 3+ promotes osteoblast activity through the phosphorylation of F AK and reorganization of the cytoskeleton.     

IRS1在藻蓝蛋白介导的H460细胞增殖和迁移调控中的作用

藻蓝蛋白来源于海洋藻类,是我国认可的食品着色剂和功能型食品．初期研究表明,藻蓝蛋白处理能抑制人类非小细胞肺癌系H460的体外增殖能力和迁移能力,使得其体外集落形成能力减弱．通过转录组学测序分析进一步探究具体作用机制,从藻蓝蛋白处理前后发生显著变化的基因中,筛选出了一个藻蓝蛋白处理后发生显著下调的基因,即胰岛素受体底物1（irs1）,并通过体外转染siRNA的方法抑制IRS1的表达,来研究其对非小细胞肺癌系H460的增殖和迁移能力的影响．采用MTT法检测细胞增殖,细胞划痕实验检测细胞迁移,克隆形成实验检测细胞集落形成能力,流式细胞术检测细胞周期分布．结果表明,下调IRS1的表达后,与对照组相比,细胞的生长速率降低,迁移能力、集落形成能力受到抑制,同时使细胞周期被阻滞到G1期．PI3K-AKT信号通路研究表明,藻蓝蛋白处理使得PI3K-AKT信号通路活性受到抑制,下调IRS1的表达使得PI3K-AKT信号通路部分蛋白表达也下调,通路活性受到一定程度抑制．本研究结果表明,藻蓝蛋白抑制非小细胞肺癌系H460体外活性功能的机制,可能与IRS1的表达和PI3K-AKT信号通路的活性有关,这为藻蓝蛋白的调控机制提供了有力的理论基础．