芦荟大黄素对结肠癌细胞株SW620 FasL基因表达和侵袭能力的影响

目的：在体外通过细胞培养，初步研究芦荟大黄素对人结肠癌SW620细胞FasLmRNA表达及对其侵袭能力的影响，为中药抗肿瘤提供实验依据。方法：根据MTT法得到芦荟大黄素对SW620细胞的半数有效抑制浓度（IC50），确定药物作用浓度．SW620细胞分别经芦荟大黄素不同浓度（0．5IC50、IC50）作用后，应用逆转录-聚合酶链反应（RT—PCR）法检测芦荟大黄素作用前后人结肠癌SW620细胞FasL mRNA的变化；应用Transwell细胞侵袭试验检测芦荟大黄素对SW620细胞侵袭能力的影响。结果：芦荟大黄素处理后较处理前SW620细胞FasLmRNA表达水平均明显高于对照组；而且FasLmRNA表达水平随芦荟大黄素作用浓度增加显著上调，芦荟大黄素不同浓度组比较均有显著性差异；随芦荟大黄素作用浓度升高，SW620细胞侵袭能力明显增强，不同浓度组比较均有显著性差异。结论：芦荟大黄素在一定时间内均可上调人结肠癌SW620细胞FasL mRNA的表达，而且这种上调作用在一定范围内呈剂量依赖性，可使结肠癌细胞的侵袭能力增强。     

阿苯达唑抑制结肠癌SW480细胞的侵袭和迁移能力及其可能的机制

目的：探索阿苯达唑（albendazole）抑制结肠癌SW480细胞侵袭和迁移能力及其可能的机制。方法：体外培养人结肠癌SW480细胞,用不同质量浓度(0、1.0和2.0 mg/ml)的albendazole处理人结肠癌SW480细胞,CCK-8法检测albendazole对结肠癌SW480细胞增殖能力的影响,采用细胞划痕实验、Transwell小室实验检测细胞的迁移侵袭能力,免疫细胞化学及Western blotting检测SW480细胞中E-cadherin、MMP-2和MMP-9蛋白的表达水平。结果：与空白对照组比较,albendazole各质量浓度（1.0和2.0 mg/ml）组细胞的增殖能力明显降低,SW480细胞侵袭细胞数显著下降\[ （51.33±3.96）、（23.42±403）vs （80.76±7.18）个/视野,F=3.975, P=0.026\];而且细胞迁移能力显著下降\[（9.6±1.13）、（6.4±0.81）vs （19.6±1.41） mm;F=5.012, P=0.023\];E-cadherin蛋白表达水平上调,MMP-2、MMP-9蛋白表达水平明显下调。结论：Albendazole能显著抑制SW480细胞增殖,并通过上调E-cadherin的表达水平和下调MMP-2和MMP-9的分泌水平抑制细胞的侵袭和迁移能力。     

NDRG2基因对结肠癌细胞SW620增殖和侵袭力的影响

目的 观察N-myc下游调节基因2(NDRG2)对结肠癌细胞SW620生长和侵袭能力的影响,并探讨其机制.方法 采用阳离子脂质体转染方法,分别将pcDNA3.1-NDRG2和siRNA-NDRG2转染入SW620细胞内,以空白组作为对照.Western blotting检测各组细胞NDRG2以及基质金属蛋白酶-2(MMP-2)的表达情况;细胞侵袭试验对各组细胞侵袭能力进行分析;四甲基偶氮唑蓝法对各组细胞生长曲线进行测定.结果 pcDNA3.1-NDRG2转染入SW620细胞后,NDRG2蛋白表达升高,而MMP-2蛋白表达降低;siRNA-NDRG2转染入SW620细胞后,NDRG2蛋白表达降低,而MMP-2蛋白表达升高.pcDNA3.1组的穿膜细胞数(56.20±7.40)及siRNA组穿膜细胞数(94.20 ±9.23)分别与对照组(75.80 ±4.82)相比,差异具有统计学意义(t=13.102,P=0.000;t=11.820,P=0.000).生长曲线显示,转染后第5天,pcDNA3.1组细胞吸光度值(0.46 ±0.01)及siRNA组细胞吸光度值(0.91 ±0.02)分别与对照组(0.67 ±0.01)相比,差异具有统计学意义(t=9.561,P=0.000;t=10.922,P=0.000).结论 NDRG2能降低结肠癌细胞SW620的侵袭和增殖能力,其机制可能与下调MMP-2的表达有关.     

肝细胞生长因子对人结肠癌细胞系内皮生长因子表达影响的观察

目的:探讨人重组肝细胞生长因子(rhHGF)对结肠癌细胞SW480和SW620中3种内皮生长因子VEGF-A、VEGF-C和VEGF-D表达的影响.方法:实验分对照组(5%胎牛血清)和HGF组(80 ng/mL的HGF).采用MTT法分析HGF对肿瘤细胞增殖的作用,流式细胞术检测HGF对细胞周期的影响.48 h后进行蛋白质印迹法检测,并对信号强度进行相对定量分析.结果:HGF能促进SW480和SW620增殖.流式细胞术结果显示,与对照组相比,SW480的HGF组中的G0/G1期细胞减少,S期细胞增多,G2/M期细胞增多,但HGF对SW620细胞周期的影响不明显.SW480细胞HGF组的VEGF-A表达量是对照组的1.59倍,SW620细胞HGF组的VEGF-A表达量是对照组的2.08倍(P＜0.01);SW480细胞HGF组的VEGF-C表达量是对照组的1.37倍,SW620细胞HGF组的VEGF-C表达量是对照组的1.27倍(P＜0.01);SW480细胞HGF组的VEGF-D表达量是对照组的1.46倍,SW620细胞HGF组的VEGF-D表达量是对照组的1.38倍(P＜0.01).结论:HGF通过上调结直肠癌细胞VEGF-A、VEGF-C和VEGF-D的表达而促进肿瘤血管和淋巴管的新生,可能是潜在的结直肠癌治疗的新靶点.     

Heparanase表达下调对结肠癌细胞增殖和侵袭的影响

目的 探讨乙酰肝素酶（Heparanase）对结肠癌细胞生物学功能的影响。方法 采用荧光定量PCR检测人结肠癌细胞株SW620、SW480、LoVo和CACO 2中Heparanase的表达水平;设计并合成3条Heparanase特异性的小分子干扰RNA（siRNA1、siRNA2、siRNA3）,用于转染4株结肠癌细胞中Heparanase表达水平最高者;荧光定量PCR和Western blotting检验3条siRNA的干扰效果;MTT法检测结肠癌细胞的增殖能力;Transwell小室法检测结肠癌细胞的侵袭能力。结果 SW480细胞中Heparanase的表达水平高于其他3株结肠癌细胞;3条特异性siRNA转染SW480细胞后,均能在蛋白和mRNA水平抑制Heparanase的表达,以siRNA3的效果最佳,且呈剂量依赖性。siRNA3介导的Heparanase基因沉默可呈剂量依赖地降低SW480细胞的增殖能力并减弱侵袭能力（P＜0.05）。结论 Heparanase表达下调可导致结肠癌细胞生物学恶性程度降低,提示Heparanase可作为结肠癌靶向治疗的一个重要靶点。     

联合转染PTEN与PINCH siRNA对结直肠癌细胞增殖、侵袭及凋亡影响的机制研究

目的:研究联合转染PTEN与PINCH siRNA对结直肠癌SW620细胞增殖、侵袭和凋亡的影响。方法:在结直肠癌SW620细胞中转染pcDNA-PTEN或/和si-PINCH，qRT-PCR检测PINCH和PTEN mRNA的表达，Western blot检测PINCH、PTEN、CDK1、MMP-2、Bcl-2和 Bax蛋白表达， MTT法、Transwell实验和流式细胞术分别检测细胞增殖、侵袭能力和凋亡率。结果:在结直肠癌SW620细胞中联合转染pcDNA-PTEN和si-PINCH后，PTEN的mRNA和蛋白表达量显著升高（P＜0.05），PINCH的mRNA和蛋白表达量显著降低（P＜0.05）；转染pcDNA-PTEN或/和si-PINCH均可抑制SW620细胞增殖和侵袭并促进细胞凋亡，抑制SW620细胞内增殖蛋白CDK1、侵袭蛋白MMP-2和抗凋亡蛋白Bcl-2的表达，促进凋亡蛋白Bax的表达；联合转染pcDNA-PTEN和si-PINCH比单独转染pcDNA-PTEN或si-PINCH效果更显著。结论:联合转染pcDNA-PTEN和si-PINCH可抑制结直肠癌SW620细胞增殖和侵袭并促进细胞凋亡，且比单独转染效果更显著。     

膜联蛋白A2对人结肠癌细胞SW620侵袭迁移的影响

目的:研究膜联蛋白A2(ANXA2)对人结肠癌细胞SW620侵袭迁移的影响.方法:采用siRNA技术沉默ANXA2在SW620细胞中的表达,实验分为干扰组、对照组和野生组,RT-PCR法检测各组ANXA2mRNA水平,Westem blot法检测各组ANXA2、纤维蛋白溶酶(PL)、基质金属蛋白酶-1(MMP-1)、血管内皮生长因子(VEGF)蛋白的表达,Transwell侵袭实验检测各组SW620细胞穿膜能力.结果:干扰组与对照组、野生组相比,ANXA2 mR-NA水平、ANXA2、PL、MMP-1、VEGF蛋白表达及SW620穿膜细胞数差异均有统计学意义(P＜0.05),而对照组和野生组相比上述各指标均无统计学差异.ANXA2蛋白和PL、MMP-1、VEGF蛋白水平、穿膜细胞数呈正相关(P＜0.05).结论:沉默ANXA2基因后可能通过下调PL、MMP-1及VEGF蛋白的表达,抑制SW620细胞侵袭迁移能力起到抗肿瘤作用.     

胶质瘤相关癌基因同源蛋白2促进结肠癌细胞系SW620的上皮-间质转化

背景与目的:胶质瘤相关癌基因同源蛋白2(glioma-associated oncogene homologue 2,GLI2)是Hedgehog信号通路重要的转录因子,不仅参与正常的细胞分化,而且在多种肿瘤细胞中异常激活,与肿瘤转移密切相关。探讨GLI2与结肠癌细胞系SW620上皮-间质转化(epithelial-mesenchymal transition,EMT)的关系及其可能机制。方法:采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)检测人结肠癌细胞系SW620、SW480、HCT116和HT29中E-cadherinmRNA表达,以GLI2干扰慢病毒感染SW620细胞,用RTFQ-PCR和蛋白质印迹法(Western blot)检测细胞中GLI2 mRNA表达和蛋白水平,采用Transwell小室检测细胞侵袭、迁移能力,黏附实验检测同、异种细胞间黏附能力,Western blot检测细胞中p-AKT、N-cadherin、vimentin、MMP2和E-cadherin蛋白水平。结果:4种细胞系中,SW620的E-cadherin mRNA表达最低(P<0.05),SW620转染72 h后,可见明显的荧光表达;与空病毒组和对照组相比,干扰组细胞的GLI2表达降低(P<0.05);细胞侵袭和迁移能力减弱(P<0.05);同种细胞黏附能力增强(P<0.05);异种细胞黏附能力减弱(P<0.05);p-AKT、N-cadherin、vimentin和MMP-2的表达降低(P<0.05)、E-cadherin表达增加(P<0.05)。结论:GLI2可能通过上调p-AKT、N-cadherin、vimentin和MMP-2表达和抑制E-cadherin表达来促进结肠癌细胞系SW620的EMT。     

肝窦内皮细胞与结肠癌细胞相互作用促进肝转移的影响

目的:探讨结肠癌细胞和肝窦内皮细胞共培养对结肠癌细胞功能以及肝转移能力的影响,为结肠癌肝转移机制的研究提供一种基本实验材料.方法:体外将人肝窦内皮细胞和结肠癌细胞进行共培养21轮,采用Transwell法、明胶酶谱分析、CCK-8法、集落形成实验检测共培养后结肠癌细胞SW1116P21体外侵袭、增殖、克隆形成能力;采用皮下成瘤实验以及实验性肝转移实验检测SW1116P21皮下成瘤以及肝转移的影响;Western blot的方法检测相关分子的改变.结果:共培养后的结肠癌细胞SW1116P21的细胞间界限明显.侵袭检测发现,SW1116P21细胞的侵袭能力较SW1116亲本细胞提高了2倍;明胶酶谱分析检测发现,SW1116P21分泌MMP-2/9的能力显著高于亲本细胞.皮下成瘤实验发现,SW1116P21皮下成瘤能力显著增强.实验性肝转移发现,SW1116P21肝转移能力显著高于SW1116细胞.Western blot检测发现SW1116P21细胞表达E-cadherin显著下调,vimentin表达上调.结论:结肠癌细胞和肝窦内皮细胞相互作用可促进瘤细胞侵袭、增殖、克隆形成以及体内细胞生长,更易发生肝转移,这种影响是与内皮细胞相互作用后结肠癌细胞发生EMT有关.     

中药夏枯草对结肠癌细胞FasL基因表达和侵袭能力的影响

目的:研究夏枯草对人结肠癌SW480细胞FasLmRNA表达及对其侵袭能力的影响,为中药抗肿瘤提供实验依据.方法: 根据MTT法得到夏枯草对SW480细胞的半数有效抑制浓度(IC50),确定药物作用浓度.SW480细胞分别经夏枯草不同浓度(0.5IC50、IC50)作用后,应用逆转录-聚合酶链反应(RT-PCR)法检测夏枯草作用前后人结肠癌SW480细胞FasL mRNA的变化;应用Transwell细胞侵袭试验检测夏枯草对SW480细胞侵袭能力的影响.结果: 夏枯草处理后较处理前SW480细胞FasL mRNA表达水平均明显高于对照组(P<0.01);而且FasL mRNA表达水平随夏枯草作用浓度增加显著上调,夏枯草不同浓度组比较均有显著性差异(P<0.01);随夏枯草作用浓度升高,SW480细胞侵袭能力明显增强,不同浓度组比较均有显著性差异(P<0.01).结论: 夏枯草在一定时间内均可上调人结肠癌SW480细胞FasL mRNA的表达,而且这种上调作用在一定范围内呈剂量依赖性,可使结肠癌细胞的侵袭能力增强.     

Effects on Proliferation and Migration of the Human Colon Carcinoma Cell Line SW620 by Silencing of Hepatocyte Growth Factor Expression

OBJECTIVE Hepatocyte growth factor (HGF) expression is closely related to the progression and poor prognosis of colorectal cancer patients. In this study, we investigated the effects on proliferation and migration of the human colon carcinoma cell line SW620 by silencing HGF expression. METHODS HGF was silenced using specifi c HGF α/β siRNA.The proliferation, migration, cell cycle and ultrastructure of SW620 cells were examined. RESULTS The transfection efficiency was 70%–80%. The expression rate of HGF in the experimental group was signifi cantly lower than that in the negative and blank control groups (P <0.05). The proliferation inhibition rate in the experimental group at 24, 48, 72 and 96 h after transfection was 14.2%, 50.2%, 39.5% and 23.2%, respectively. The migratory ability of cells in the experimental group was significantly inhibited compared with that in the negative control or blank control groups (58.2% vs. 2.1% or 0%, P < 0.05).CONCLUSION The application of RNA interference to silence the expression of HGF in the colon carcinoma cell line SW620 effectively inhibits the proliferation and migration of tumor cells.     

二氢卟吩e6光动力对人结肠癌细胞SW620生物学行为的影响

目的探讨二氢卟吩e6(Ce6)光动力学疗法(PDT)对人结肠癌SW620细胞增殖、迁移及克隆形成能力等生物学行为的影响。方法Ce6光动力处理SW620细胞,采用MTT法检测细胞增殖,划痕实验检测细胞迁移能力,平板克隆形成实验检测细胞克隆形成能力。结果Ce6浓度和光照剂量增加时,Ce6-PDT对SW620细胞生长抑制作用逐渐增加,呈现浓度依赖、光照剂量依赖关系。划痕实验结果显示Ce6-PDT能够抑制细胞的迁移。Ce6-PDT能够减弱SW620细胞克隆形成能力。结论Ce6-PDT能够抑制SW620细胞增殖、迁移及克隆形成能力。     

Effects on proliferation and migration of the human colon carcinoma cell line SW620 by silencing of hepatocyte growth factor expression

Objective  

Hepatocyte growth factor (HGF) expression is closely related to the progression and poor prognosis of colorectal cancer patients. In this study, we investigated the effects on proliferation and migration of the human colon carcinoma cell line SW620 by silencing HGF expression.     

EGCG对因子Ⅶa促进SW620细胞增殖与迁移的干预作用及机制

目的探讨表没食子儿茶素没食子酸酯(EGCG)对因子Ⅶa促进SW620细胞增殖与迁移的干预作用及机制。方法用EGCG、因子Ⅶa等处理SW620细胞,应用流式细胞术、Transwell法分别检测细胞增殖周期及迁移能力;Western blotting检测细胞NF-κB抑制蛋白(IκB-α)、核NF-κB(p65/RelA)的蛋白水平变化。结果 EGCG(100 mg/L)能干预因子Ⅶa对SW620细胞增殖与迁移的促进作用;EGCG能抑制因子Ⅶa对IκB-α表达的下调作用及对核NF-κB(p65/RelA)表达的促进作用。结论 EGCG可能通过干预信号分子IκB-α和NF-κB(p65/RelA)的表达和调节作用,抑制因子Ⅶa对SW620细胞增殖与迁移的促进作用。     

TF/FVIIa/PAR2 promotes cell proliferation and migration via PKCα and ERK-dependent c-Jun/AP-1 pathway in colon cancer cell line SW620

Our previous study has demonstrated that tissue factor–factor VIIa (TF/FVIIa) complex promotes the proliferation and migration of colon cancer cell line SW620 through the activation of protease-activated receptor 2 (PAR2). In the current study, the underlying molecular mechanisms of TF/FVIIa/PAR2 signaling in SW620 cells were further explored, with the focus on the role of activator protein-1 (AP-1) subunit c-Jun. The results revealed that PAR2-AP and FVIIa could upregulate c-Jun expression and c-Jun phosphorylation in SW620 cells in a time-dependent manner. The effect of FVIIa was significantly blocked by anti-TF and anti-PAR2 antibodies. Protein kinase Cα (PKCα) inhibitor safingol and extracellular signal-regulated kinase 1 and 2 (ERK1/2) inhibitor U0126 abrogated the activation of c-Jun. In contrast, Ca²⁺ chelators EGTA and thapsigargin, and p38MAPK inhibitor SB203580 had no effect. Suppression of c-Jun/AP-1 activation using a natural inhibitor curcumin decreased the expression of caspase-3, MMP-9, and TF, as well as the proliferation and migration of SW620 cells induced by PAR2-AP or FVIIa. Collectively, our findings suggest that c-Jun/AP-1 activation is required for TF/FVIIa/PAR2-induced SW620 cell proliferation and migration. PKCα and ERK1/2 are located upstream of c-Jun/AP-1 in this signaling pathway. Pharmacological inhibition of this pathway might be a novel strategy for colon cancer therapy.     

Involvement of PKCα activation in TF/VIIa/PAR2-induced proliferation, migration, and survival of colon cancer cell SW620

Our previous study has demonstrated that protease-activated receptor 2 (PAR2) activation mediated by tissue factor (TF)/VIIa complex triggers the ERK1/2/NF-κB signaling pathway, which further contributes to the proliferation and migration of colon cancer cell line SW620. However, the detailed mechanisms remain unclear. This study was to investigate whether protein kinase Cα (PKCα) is involved in these events and the possible mechanism. The results revealed that PAR2-activating peptide or VIIa could induce time-dependent upregulation of PKCα phosphorylation in SW620 cells and PKCα translocation from the cytoplasm to the perinuclear region and nucleus. The activation of PKCα was sufficient to induce ERK1/2 and NF-κB phosphorylation. The VIIa effect was obviously blocked by both anti-TF and anti-PAR2 antibodies. The PKCα inhibitor, safingol, inhibited ERK1/2 phosphorylation and NF-κB activation that is induced by VIIa and abrogated the enhanced proliferation, migration, and survival of SW620 cells by VIIa treatment. Both safingol and PDTC (NF-κB inhibitor) could apparently rescue the effects of VIIa on expression of MMP-9, caspase-3, TF, and Bcl-2/bax in SW620 cells. Collectively, the data in this study suggest that TF/VIIa/PAR2-induced SW620 cell proliferation, migration, and survival are ascribed to the activation of PKCα, and these effects are achieved through PKCα downstream signaling pathways, ERK1/2 and NF-κB.     

The expression and the functional roles of tissue factor and protease-activated receptor-2 on SW620 cells

Tissue factor (TF) is believed to play an important role in tissue repair, inflammation, angiogenesis, and tumor metastasis. Protease-activated receptors (PARs) are widely expressed on various cells including tumor cells and associated with many pathological mechanisms. In the present study, the expression of TF and PAR1, PAR2 on human colon cancer cells (SW620 and SW480) was investigated and their functional roles on the behavior of tumor cells were evaluated. It was demonstrated that SW620 and SW480 cells expressed TF at antigen, activity and mRNA levels. However, the highly metastatic cell line SW620 showed slightly higher TF expression than the low metastatic cell line SW480. The PAR2 antigen was strongly expressed on the membrane of SW620 cells, but not on SW480 cells. The PAR1 antigen was not observed in SW620 or SW480 cells, while PAR1 and PAR2 mRNA was detected in SW620 and SW480 cells. The migratory potential of SW620 was stronger than that of SW480 seen in Boyden chambers. PAR2 agonist (SLIGKV-NH2) and factor VIIa significantly stimulated SW620 cell proliferation, migratory activity, and interleukin 8 (IL-8) secretion compared to control. The stimulating effects of factor VIIa could be inhibited by anti-TF and anti-PAR2 but not anti-PAR1 antibodies. In summary, this study demonstrates that TF and PAR2 are strongly expressed on highly metastatic colonic tumor cells and are closely associated with the proliferation and migration of the cells. TF may elucidate its roles in colonic cancer invasion and metastasis via PAR2 pathway.     

Activation of PAR2 or/and TLR4 promotes SW620 cell proliferation and migration via phosphorylation of ERK1/2

Protease-activated receptor 2 (PAR2) is a G-protein-coupled receptor and its activation has been associated with the pathogenetic progress in certain cancers. Toll-like receptor 4 (TLR4), one member of Toll-like receptors family, is mainly contributed to the innate immune response. However, recent studies have shown that TLR4 is aberrantly expressed in various types of carcinomas and may correlate with tumor progression. Previously, we reported that PAR2 could be expressed in human colon cancer cell line (SW620) and its activation by some stimulants was able to facilitate cell proliferation and migration. In our recent preliminary experiment, it was found that SW620 cells also had TLR4 expression. Thus, we considered that PAR2 and TLR4 could have some collaborative roles in SW620 cells. In the current study, the cross-inducible expression of PAR2 and TLR4 on SW620 cells was investigated, and the functional roles of their activation on the behavior of SW620 cells were evaluated. It was found that activation of PAR2 with PAR2-AP (PAR2 agonist, 100 μM) enhanced TLR4 releasement and vice versa. The activation of PAR2 or TLR4 (with LPS, 100 ng/ml) could promote SW620 cell proliferation and migration, and the phosphorylation of ERK1/2 but not p38MAPK, as well as the expression of interleukin 8 (IL-8) and tissue factor (TF). Whereas the caspase-7 expression was decreased under PAR2 or TLR4 activation. Furthermore, ERK1/2 inhibitor (U0126, at 10 μM) could intervene in all regulating effects of PAR2 or/and TLR4. Collectively, this study demonstrated that both PAR2 and TLR4 activation on SW620 cells can trigger the phosphorylation of ERK1/2, regulate the expression of IL-8, TF and caspase-7, thereby promote the proliferation and migration of cells.     

过表达血管生成抑制蛋白1对人结直肠癌细胞恶性生物学行为的影响

目的： 探讨过表达血管生成抑制蛋白1（vasohibin-1,VASH1）对人结直肠癌细胞恶性生物学行为的影响。方法： 包装慢病毒并感染人结直肠癌SW680、SW620细胞以构建过表达VASH1的细胞系,以未经感染的细胞为对照;qPCR实验和WB实验检测VASH1的过表达效果,小管形成实验、CCK-8实验、软琼脂克隆形成实验、Transwell实验和划痕愈合实验检测过表达VASH1在体外对细胞的微血管形成、增殖、克隆形成以及迁移能力的影响,NOD-SCID小鼠皮下成瘤实验检测过表达VASH1对SW620细胞移植瘤的体内生长和肺转移的影响。结果： 成功构建过表达VASH1的SW480和SW620细胞。体外实验表明,与对照组相比,过表达VASH1的结直肠癌细胞的微血管形成能力、增殖能力、克隆形成能力以及迁移能力均显著降低（均P<0.05）。体内实验表明,与对照组相比,过表达VASH1的SW620细胞NOD-SCID小鼠皮下移植瘤的生长速度以及肺转移能力也明显降低（均P<0.05）。结论： 过表达VASH1能够抑制人结直肠癌细胞的恶性生物学行为。     

核因子κB在凝血因子Ⅶa促进结肠癌SW620细胞增殖迁移中的作用

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。