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1.
目的 体外探讨凝血因子Ⅶa促进结肠癌细胞株SW620增殖与迁移的作用机制.方法 采用蛋白酶激活受体2激动剂(PAR2-AP)、凝血因子Ⅶa等处理SW620细胞,以实时定量PCR检测SW620细胞中白细胞介素8(IL-8)、组织因子(TF)及半胱氨酸蛋白酶7(caspase-7)mRNA的表达水平;以酶联免疫吸附试验(ELISA)检测细胞上清IL-8蛋白的含量;以Xa生成法检测细胞TF活性;以Western blot法检测细胞磷酸化p38丝裂原活化蛋白激酶(p-p38 MAPK)水平.结果 PAR2-AP、凝血因子Ⅶa能够增加SW620细胞中IL-8基因和蛋白的表达,上调TF mRNA水平及活性,下调caspase-7基因表达和p-p38 MAPK水平,单克隆抗TF及抗PAR2抗体均可抑制凝血因子Ⅶa的作用.结论 凝血因子Ⅶa与细胞表面TF形成复合物,通过活化PAR2,上调结肠癌细胞株SW620中IL-8、TF的表达,下调细胞caspase-7表达,从而促进细胞增殖与迁移能力,p38 IVIAPK在此过程中起负性调节作用. 相似文献
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目的 探讨核因子κB(NF-κB)在促进结肠癌SW620细胞增殖迁移过程中的作用及其可能的机制。方法 以凝血因子Ⅶa、NF-κB抑制剂二硫代氨基甲酸吡咯烷(PDTC)等处理结肠癌细胞株SW620,采用Western blot法检测细胞核NF-κB(p65)、细胞浆NF-κB抑制蛋白(IκB-o)和凋亡蛋白半胱氨酸蛋白酶7(caspase-7)的蛋白表达变化;采用流式细胞术检测SW620细胞的细胞周期变化;采用Transwell法测定SW620细胞的迁移能力;采用实时定量聚合酶链反应(PCR)检测白细胞介素8( IL-8)和组织因子(TF) mRNA的表达水平。结果凝血因子Ⅶa能够显著下调细胞浆中IκB-o的表达水平,并使细胞核内NF-κB的表达水平升高,单克隆抗TF和抗蛋白酶激活受体2(PAR2)抗体能够抑制凝血因子Ⅶa的这一作用。PDTC能够明显干预凝血因子Ⅶa对SW620细胞增殖、迁移的促进作用。PDTC能够明显干预凝血因子Ⅶa对SW620细胞中TF、IL-8 mRNA表达的促进作用和对caspase-7蛋白表达的下调作用。结论 凝血因子Ⅶa与细胞表面TF结合形成TF/Ⅶa复合物,活化受体PAR2,经NF-κB通路上调IL-8、下调caspase-7的表达,促进SW620细胞的增殖与迁移。TF/Ⅶa/PAR2/NF-κB通路还可进一步上调TF的表达,从而形成TF/Ⅶa/PAR2/NF-κB/TF正反馈通路。 相似文献
3.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
4.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
5.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
6.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
7.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
8.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
9.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
10.
Objective To investigate the mechanisms that coagulation factor Ⅶa promotes proliferation and migration of a colon cancer cell line (SW620 cells) in vitro. Methods The expression of interleukin 8 (IL-8), tissue factor (TF), caspase-7 and p-p38 MAPK in SW620 cells treated with factor Ⅶa or protease activated receptor 2 aganist (PAR2-AP) was measured by ELISA, Western-blotting and QT-PCR. Results Factor Ⅶa and PAR2-AP induced IL-8 expression at both mRNA and protein levels, up-regulated TF mRNA expression and TF activity, but down-regulated caspese-7 mRNA and p-p38 MAPK levels in SW620 cells. The effects of factor Ⅶa were not only blocked by anti-TF but also by anti-PAR2 antibodies. Conclusion Factor Ⅶa binds to TF on cell surface, forming a complex which activates PAR2, then provoking IL-8 and TF expression, and suppresses caepase-7 expression, thus promotes the tumor cell proliferation and migration, p38 MAPK may negatively regulate this process. 相似文献
11.
背景与目的探讨绿茶提取物表没食子儿茶素没食子酸酯(epigallocatechin-3-gallate,EGCG)诱导人胃癌细胞裸鼠移植瘤凋亡及分子机制。方法建立人胃腺癌(SGC7901)细胞裸鼠异种移植瘤模型,用不同剂量的EGCG进行治疗,并设对照,用流式细胞分析术检测肿瘤组织中细胞凋亡情况,免疫组织化学方法检测肿瘤组织的凋亡相关基因Bcl-2、Bax、Caspase-3的表达情况。结果裸鼠异种移植瘤治疗实验结果显示,EGCG对移植瘤生长有明显抑制作用(其中20mg/kg、EGCG抑制率54.64%与对照组比较P<0.05);PI染色FCM分析发现,EGCG20mg/kg能诱导移植瘤细胞凋亡率17.2%与对照组比P<0.05;SP免疫组织化学结果表明EGCG可上调移植瘤细胞中Bax、Caspase-3蛋白的表达,下调Bcl-2蛋白的表达。结论表没食子儿茶素没食子酸酯(EGCG)具有体内诱导人胃腺癌细胞凋亡的作用,其机制可能与Bcl-2/Bax比值降低导致Caspase-3的活化从而启动细胞凋亡有关。 相似文献
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目的: 了解玻连蛋白(VTN)对肝肿瘤细胞株SMMC7721增殖和迁移能力的影响。方法: 采用不同浓度VTN包被培养皿后接种、培养细胞,采用激光共聚焦显微镜观察VTN包被后细胞骨架成分微管蛋白形态的变化,用细胞增殖试剂分析VTN对细胞增殖率的影响及分析VTN对凋亡诱导剂抑制增殖作用的影响,采用Transwell侵袭实验检测VTN对细胞迁移能力的影响。结果:激光共聚焦观察结果显示VTN包被有助于促进细胞贴壁及细胞骨架蛋白形态结构的维持;VTN可促进肿瘤细胞的生长且呈剂量效应,并可减弱凋亡诱导剂对细胞增殖的抑制;Transwell侵袭实验结果显示VTN包被可促进细胞迁移。结论:体外实验条件下,VTN有助于肝癌细胞株SMMC7721的增殖及迁移,提示VTN在肝癌发生过程中可能具有促进肿瘤细胞增殖和迁移的生物学作用。 相似文献
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目的:研究瘦素对乳腺癌细胞株MCF-7细胞生长的作用。方法:分别使用0、25、50、100和200nmol/L瘦素作用于细胞株MCF-7细胞,同时在上述作用后24、48和72h采用光学显微镜观察细胞形态学改变,采用MTT比色方法,观察不同浓度瘦素对MCF-7细胞生长刺激的剂量、时间效应,比较在相对应剂量胰岛素情况下,二者对细胞增殖刺激作用的强弱。结果:加用瘦素各组细胞比未加瘦素细胞密度增加,形态变异。与正常对照组相比,各浓度瘦素(25、50、100和200nmol/L)均可明显促进MCF-7细胞增殖,其中50、100和200nmol/L瘦素作用24、48和72h后差异均有统计学意义,P〈0.05。瘦素对MCF-7细胞的增殖作用呈现一定的时间和剂量依赖效应。胰岛素各相对应组之间与瘦素比较,细胞增殖刺激作用差异无统计学意义,P〉0.05。结论:瘦素对乳腺癌细胞株MCF-7细胞增殖有刺激作用,且这种作用呈现一定的剂量、时间依赖效应,结果提示瘦素是促进乳腺癌细胞增殖的因素之一。 相似文献
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目的:研究瘦素对乳腺癌细胞株MCF-7细胞生长的作用。方法:分别使用0、25、50、100和200nmol/L瘦素作用于细胞株MCF-7细胞,同时在上述作用后24、48和72h采用光学显微镜观察细胞形态学改变,采用MTT比色方法,观察不同浓度瘦素对MCF-7细胞生长刺激的剂量、时间效应,比较在相对应剂量胰岛素情况下,二者对细胞增殖刺激作用的强弱。结果:加用瘦素各组细胞比未加瘦素细胞密度增加,形态变异。与正常对照组相比,各浓度瘦素(25、50、100和200nmol/L)均可明显促进MCF-7细胞增殖,其中50、100和200nmol/L瘦素作用24、48和72h后差异均有统计学意义,P<0.05。瘦素对MCF-7细胞的增殖作用呈现一定的时间和剂量依赖效应。胰岛素各相对应组之间与瘦素比较,细胞增殖刺激作用差异无统计学意义,P>0.05。结论:瘦素对乳腺癌细胞株MCF-7细胞增殖有刺激作用,且这种作用呈现一定的剂量、时间依赖效应,结果提示瘦素是促进乳腺癌细胞增殖的因素之一。 相似文献
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目的 研究蜂胶黄酮(pinobanksin-3-acetate,PB3A)对体外培养的大肠癌SW480细胞中G蛋白信号转导调节因子2 (regulator of G-protein signaling 2,RGS2)和GTP结合丝裂原诱导蛋白(GTP binding protein overexpressed in skeletal muscle,GEM)基因转录和蛋白质表达水平的影响,探讨PB3A的抗肿瘤作用机制及其相关信号通路.方法 通过实时荧光定量RT-PCR和蛋白印迹法检测100 mg/L PB3A作用下人大肠癌细胞SW480中RGS2和GEM基因的转录和表达情况.结果 100 mg/L PB3A药物的干预诱导SW480细胞出现悬浮的细胞碎片、细胞体缩小、变圆、皱缩.药物干预之后,SW480细胞中RGS2和GEM基因在mRNA和蛋白水平上调表达.实验组与对照组比较,RGS2和GEM基因的mRNA转录水平差异有统计学意义(P<0.01),蛋白质表达水平差异有统计学意义(P<0.05).根据RT-PCR检测结果,利用Spearman相关性分析检测出RGS2和GEM高度正相关(r=0.929,P<0.01).结论 PB3A干预大肠癌SW480细胞后使RGS2和GEM基因在转录和翻译水平上上调表达.PB3A的抗大肠癌作用机制可能与RGS2和GEM基因的上调表达有关. 相似文献
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EGCG对人乳腺癌MDA-MB-435细胞株细胞增殖的抑制及机制 总被引:8,自引:0,他引:8
目的:研究绿茶提取物表没食子儿茶素—3—没食子酸酯(epigalloeateehin—3 gallste,EGCG)对人乳腺癌细胞株细胞周期的影响及机制。方法:利用细胞增殖测定试剂盒(Cell Proliferation Assay kit)来制作细胞生长曲线;流式细胞仪分析EGCG作用于人乳腺癌MDA—MB—435细胞株前后细胞周期的变化;RT—PCR及Western印迹法研究肿瘤细胞株加药干预前后细胞周期抑制因子p21^waf1/cip1的mRNA及蛋白表达的变化。结果:EGCG可明显抑制乳腺癌细胞MDA—MB—435细胞的增殖活性,40μg/ml EGCG干预后,MDA-M-B435细胞主要阻滞于G0/G1期,在24小时阻滞最为明显。在48及72小时也有阻滞。对照组G0/G1期的比例49.92%,40μg/ml EGCG干预24、48及72小时后G0/G1期的比例分别为71.56%、67.20%和61.59%。EGCG 40μg/ml处理肿瘤细胞株细胞后24小时可检测p21 mRNA和蛋白表达提高。结论:绿茶有效成分可抑制乳腺癌细胞的增殖,这可能与其诱导p21表达从而抑制细胞周期的时相转换有关,这表明绿茶可能在乳腺癌的治疗上具有一定的前景。 相似文献
17.
目的:研究TLR4基因多态性对人肝癌细胞HepG2细胞增殖、迁移及凋亡的影响,并探讨其相关分子机制.方法:构建TLR4野生型(WT)、1196(C/T)位点及896(A/G)位点突变型质粒并将其稳转HepG2细胞株中;CCK-8法检测、Annexin V-PE/7-AAD流式细胞仪及Transwell小室实验检测TLR4基因多态性对HepG2细胞增殖、迁移及凋亡的影响;Western blot检测NF-κB p65(nuclear factorκB p65)表达水平的差异.结果:Western blot结果表明三组TLR4过表达细胞株中TLR4的含量显著高于正常组.与正常HepG2细胞相比,三组TLR4过表达细胞株增殖能力、迁移能力及p65表达量均增加;与TLR4野生型组相比,两组突变型细胞株增殖能力、迁移能力及p65表达量均减弱,且凋亡率增加,差异具有统计学意义(P<0.05).结论:TLR4可能通过影响NF-κB p65相关蛋白的表达促进肝癌细胞HepG2的增殖及迁移,其基因1196(C/T)位点及896(A/G)位点的突变会减弱肝癌细胞的增殖及迁移能力. 相似文献
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目的分析紫杉醇脂质体对卵巢癌SKOV-3细胞的生长抑制作用,观察药物作用的时间-浓度关系,探究药物作用机制。方法应用不同剂量紫杉醇脂质体对SKOV-3细胞进行处理,通过MTT检测细胞存活率,倒置显微镜观察细胞形态,流式细胞术检测细胞凋亡作用及其对细胞周期的阻滞状态,Western blot分析Bcl-2基因蛋白表达情况。利用SPSS 13.0对数据进行统计学分析。结果紫杉醇脂质体对SKOV-3细胞有明显的剂量-时间依赖效应。紫杉醇与紫杉醇脂质体均可将SKOV-3细胞阻滞在G2/M期,且随药物浓度增加,凋亡细胞所占比例逐渐增加。通过Western blot方法发现,紫杉醇脂质体作用后SKOV-3细胞Bcl-2蛋白表达明显下降,Bax蛋白表达明显上调,Bax/Bal-2比例明显上调。结论紫杉醇以脂质体为转运载体后,并未改变其对卵巢癌SKOV-3细胞的抑制作用及作用周期,具有较好的临床应用前景。 相似文献
19.
目的 研究miR-200b对人结肠癌sw620细胞E-cadherin表达的调节作用,探讨miR-200b对sw620细胞增殖及迁移的影响.方法 采用实时PCR (RT-PCR)技术分别检测转染pEGP-miR-200b后24h和72 h sw620细胞中miR-200b的表达变化;采用RT-PCR技术和Western blot方法检测转染miR-200b后对E-cadherin表达的影响;采用四甲基偶氮唑蓝法(MTT)和划痕擦伤实验检测转染miR-200b对sw620细胞增殖、迁移的影响.结果 miR-200b转染sw620细胞48 h和72 h后,miR-200b相对表达量较对照组显著上调(t=11.579,P<0.01;t=11.579,P<0.01);且抑制了细胞的增殖能力,第3天抑制作用最显著,抑制率为55.34%;细胞的迁移速度受到明显的抑制,两组在24、48、72 h的迁移距离有显著差异(t=11.579,P<0.01;t=10.419,P<0.01;t=6.955,P<0.01).同时细胞中E-cadherin基因的mRNA和蛋白表达水平显著升高(t=10.432,P<0.01;t=8.325,P<0.01).结论 上调miR-200b表达可抑制结肠癌细胞的生物活性,该作用可能与E-cadherin的表达有关. 相似文献
20.
β肽及其多聚物对人肝癌细胞株与基质黏附的抑制作用 总被引:1,自引:0,他引:1
目的研究β肽及其多聚物对肝癌细胞株与基质黏附的抑制作用.方法观察化学合成的β肽(β肽)、化学合成的二聚β肽(β2肽)、化学合成的三聚β肽(β3肽)、用大肠杆菌表达系统表达的表达β3肽及GRGDS对人肝癌细胞株SMMC-7721细胞及人肝癌高转移细胞株HCCLM6细胞与纤连蛋白(fibronectin,FN)的黏附作用的影响.结果各种多肽对细胞与FN的黏附均具有特异的抑制作用,呈现剂量效应相关关系和时间效应相关关系(P〈0.05).且随着多肽重复次数的增多,对细胞与FN黏附的抑制作用越强,表达β3肽和β3肽的抑制肿瘤细胞黏附的作用最强,β2肽和GRGDS的作用次之,β肽的抑制作用最弱.各种多肽对HCCLM6细胞与FN黏附的抑制作用强于对SMMC-7721细胞的黏附抑制作用.结论表达β3肽对肝癌细胞株与基质的黏附具有强大的抑制作用,可能成为抗肿瘤转移的新的药物手段. 相似文献