维生素D受体基因多态性对脊柱结核易感性的影响

目的：探讨维生素D受体（vitamin D receptor，VDR）基因多态性与中国湖南省汉族人群脊柱结核易感性的关系。方法：选取2004年10月至2006年2月我院收治的湖南省汉族新发脊柱结核患者42例（病例组）及志愿者64例（对照组），应用聚合酶链反应-限制性片断长度多态性分析技术检测两组对象VDR基因FokⅠ酶切位点多态性，并进行VDR基因分型。结果：VDR—FF、VDR—Ff与VDR—ff三种基因型在病例组和对照组中的分布频率分别为14．29％、35．71％、50％和23．45％、54．69％、21．86％；两组组间比较有显著性差异（P〈0．05）．其中VDR—ff基因型在病例组中的分布频率明显高于对照组，比值比（odds ratio，OR）为3．571（P〈0．05）．其95％可信区间（confidence interval，CI）为1．561～8.167。结论：VDR基因FokⅠ酶切值点多念性与湖南省汉族人群脊柱结核的易感性相关，VDR—ff基因型可能是其易感基因型。     

维生素D受体基因多态性与冠心病的相关性

近年来国内外研究均证实维生素D能够抑制炎症反应程度,从而具有延缓冠状动脉粥样硬化的作用,而维生素D的作用通过维生素D受体来实现,现已知维生素D受体基因存在变异,而这些变异可能影响维生素D的作用。本文拟就维生素D 及其受体基因变异与动脉硬化相关心血管疾病的关系进行综述。     

3-溴丙酮酸对前列腺癌PC-3细胞增殖、迁移及侵袭的影响

目的:观察3-溴丙酮酸(3-BrPA)对前列腺癌细胞PC-3增殖、迁移和侵袭能力的影响,并初步探讨其内在机制。方法:体外培养PC-3细胞,倒置显微镜下观察不同浓度3-BrPA对PC-3细胞形态学的影响;应用MTT法、细胞划痕和Transwell法检测不同浓度3-BrPA在不同时间段(24、48、72 h)对前列腺癌PC-3细胞增殖、迁移和侵袭的影响;应用Western印迹检测3-BrPA作用后各组PC-3细胞葡萄糖转运蛋白1(GLUT1)和基质金属蛋白酶14、9、2(MMP-14、MMP-9、MMP-2)蛋白表达的变化。结果:在一定浓度范围内,随着3-BrPA浓度的升高,细胞形态的改变愈明显。MTT结果表明,随着3-BrPA浓度的增加、作用时间延长PC-3细胞抑制率增大(P<0.01)。细胞划痕和Transwell细胞侵袭实验显示:与对照组相比,培养24 h后25、50、100μmol/L 3-BrPA浓度组细胞划痕迁移愈合率降低,细胞侵袭数目减少,且培养48 h与24 h相比,各组细胞迁移愈合率和侵袭细胞数目均降低,表明细胞体外迁移率和侵袭细胞数目与3-BrPA作用浓度、作用时间有关,具有剂量和时间依赖性(P<0.01)。Western印迹检测结果表明:与对照组相比,25、50、100μmol/L 3-BrPA浓度组中GLUT1、MMP-14、MMP-9、MMP-2蛋白表达量明显降低(P<0.01)。结论:糖酵解抑制剂3-BrPA降低PC-3增殖、迁移和侵袭能力,可能与下调GLUT1、MMP-14、MMP-9及MMP-2蛋白表达有关。     

上海地区汉人维生素D受体基因多态性与骨密度关系的初步分析

目的：了解维生素D受体（VDR）基因多态性在中国人群中的分布，并进一步研究其与骨密度的关系。方法：通过聚合酶链反应－限制性片段长度多态性（PCR－RFLP）方法分析了348例无亲缘关系的上海地区男女居民的VDR基因型，并用双能X线吸收仪测定了其中202例骨密度。结果：348例研究对象中bb型占81．9％，Bb型占18．1％，未见到BB型。b等位基因在本组人群中分布 高达90．0％。男女性之间VDR基因型分布频率无明显区别（P＞0．5）。比较这两组各部位的骨密度值，只有女性在华氏三角区部位显示出Bb型比bb型有较高的BMD，在其余部位，不管男性还是女性，两组基因型的BMD均差异无显性（P＞0．05）。结论：VDR基因多态性与骨密度无相关关系。     

维生素D受体基因BsmI多态性与发育性髋关节发育不良的相关性研究

[目的]明确我国北方汉族人维生素D受体基因(vitamin dreceptor gene,VDR)Bsml位点多态性分布及与发育性髋关节发育不良(developmental dysplasia op the hip,DDH)的相关性.[方法]采用PCR-测序的方法,对54个DDH患儿及55个正常儿的VDR基因BsmI位点进行分型.采用精确概率的x2检验,比较病例组及对照组中等位基因及基因型频率分布的差异.并把正常组与文献报道的不同人群该位点的多态性分布情况相比较,以了解VDR基因BsmI位点多态性在不同种族人群中的差异.[结果]VDR基因Bsml位点的基因型及等位基因频率分布,在DDH组与对照组之间均无明显的统计学差异(P值分别为0.527和0.428);我国北方汉族人该位点的基因型以GG为主,与美国人、澳洲人及法国人均存在明显的统计学差异(P<0.01).[结论]VDR基因Bsnfl位点基因型分布存在人群差异;该位点多态性与我国北方汉族人DDH的发生可能无关.     

ShRNA靶向沉默FGFR3基因对人肝癌细胞Huh7增殖和迁移的影响

目的:研究成纤维细胞生长因子受体3(FGFR3)对肝癌细胞株Huh7增殖和迁移能力的影响。方法:以靶向沉默FGFR3的shRNA慢病毒表达载体、delta8.9和VSVG三质粒共转染293T细胞来包装慢病毒颗粒。选择Huh7细胞转导慢病毒载体。通过细胞增殖实验(CCK-8法)和transwell实验,分别评估沉默FGFR3对Huh7肝癌细胞增殖和迁移能力的影响。裸鼠分别皮下注射pSilencer-FGFR3-shRNA1#组和pSilencer-NC组Huh7细胞后,监测肿瘤生长。Western印迹法检测下游信号蛋白p-AKT和p-ERK。结果:与NC组比较,RNAi组的细胞增殖能力显著下降(P     

靶向沉默核干因子对前列腺癌PC-3细胞增殖能力的影响

目的:检测前列腺癌PC-3、LNCaP及DU145细胞中核干因子(Nucleostemin,NS)基因的表达,研究NS基因沉默后对PC-3细胞增殖能力的影响。方法:采用免疫细胞化学法及逆转录-聚合酶链反应(RT-PCR)分别检测NS蛋白及mRNA在3种前列腺癌细胞中的表达。用NS特异性小发夹RNA表达质粒转染PC-3细胞,分别用RT-PCR及Western印迹方法检测转染后细胞(简称NS-shRNA-PC-3)中NSmRNA及蛋白的变化。比较NS基因沉默前后PC-3细胞体外、裸鼠体内增殖能力及凋亡情况的变化。结果:3种细胞中均显示NS基因高表达。转染后NS-shRNA-PC-3细胞中NS表达显著降低,细胞增殖速度减慢,G0/G1期细胞百分率显著升高,早期凋亡细胞增多。体内致瘤实验显示,NS基因沉默后,PC-3细胞在裸鼠体内增殖能力显著降低。结论:NS在前列腺癌细胞系中呈高表达,RNA干扰沉默NS基因后PC-3细胞增殖能力显著降低,凋亡细胞增多。     

靶向PPARγ基因siRNA载体的构建及沉默效应

目的构建靶向过氧化物酶体增殖物激活受体-γ（peroxisome proliferator—activated receptor-γ，PPARγ）基因的小干涉RNA（small interfering RNA，siRNA）载体，转染骨髓基质细胞（marrow stromal cells，MSCs）后观察其对PPARγmRNA的抑制作用。方法根据siRNA设计原则，在PPARγmRNA序列中选择2个特异性靶序列（677-695和497-515）及同时设计1条无关对照序列；体外合成对应发卡样DNA片段，经退火后，将其定向克隆入siRNA载体pRNAT—U6．2，将重组siRNA载体pRNAT—U6．2-PPARγ用脂质体包裹转染入MSCs后，采用RT—PCR检测PPARγ基因mRNA表达水平的变化。结果得到阳性重组克隆pRNAT—U6．2-SPPARγ，经过PCR鉴定与测序，结果正确；RT—PCR检测转染MSCs显示，PPARγ基因的表达水平明显降低。结论成功构建靶向PPARγ基因的siRNA载体pRNAT—U6．2-SPPARγ，转染MSCs可以有效抑制PPARγmRNA的表达，这为预防酒精性股骨头坏死发生的研究奠定了可靠基础。     

北方汉族人群维生素D受体基因Bsm I位点多态性与前列腺癌易感性的相关性分析

目的 :研究低发病的中国汉族人群维生素D受体基因 (VDRG)BsmⅠ 位点单核苷酸多态性 (SNP)与前列腺癌的关系 ,探讨不同种族前列腺癌发病的基因差异。　方法 :收集中国北方地区汉族人群 10 3例前列腺癌病人及10 6例健康对照者外周血标本 ,应用变性高效液相色谱 (DHPLC)检测VDRG第 8内含子BsmⅠ多态位点 ,并对该位点SNP分布进行分析。　结果 :BsmⅠ 多态位点bb、Bb、BB基因型和等位基因在北方地区汉族前列腺癌病人及对照者中的分布频率差异无显著性 (P >0 .0 5 ) ,基因型分布频率分别为 92 .2 3%、7.77%、0和 94.34 %、5 .6 6 %、0 ;等位基因B、b分别为 3.88%、96 .12 %和 2 .91%、97.0 9%,而与高发病人群的分布相比有显著不同。　结论 :VDRGBsmⅠ多态性在低发病的中国汉族人群与前列腺癌无相关 ,其分布与高发病人群有明显差异 ,提示VDRGBsmⅠ多态性可能是前列腺癌发病种族差异的原因之一。     

维生素D受体基因多态性与原发性甲状旁腺功能亢进症的关系

目的 探讨维生素D受体（vitamin Dreceptor ,VDR)基因多态性与原发性甲状旁腺功能亢进症（primary hyperparathyroidism,PHPT)的关系。方法 应用多聚酶链反应法和限制性内切酶技术检测30例PHPT患者和60例正常对照组的VDR基因型。结果 VDR基因型在PHPT患者中的分布为BB型0例，Bb型1例（3．3％），bb型29例（96．7％）；正常对照组BB型2例（3．3％），Bb型11例（18．4％），bb型47例（78．3％）。PHPT患者与对照组BB，Bb,bb基因型分布差异有显著性（P≤0.05）。结论 PHPT与VDR基因多态性有一定关系。     

沉默Polo样激酶1基因的表达对结直肠癌细胞迁移和侵袭的影响

目的 研究PLK1在结直肠癌细胞迁移和侵袭过程中的作用.方法 常规培养9株结直肠癌细胞,采用PCR和Western-blot法筛选PLK1高表达细胞株.设计3种RNA干扰片段,转染高表达PLK1的结直肠癌细胞株,利用实时定量PCR和western-blot法验证并干扰效果.选择高干扰效果的干扰片段,通过Transwe1l实验来评估转染后结直肠癌细胞迁移和侵袭能力的变化.结果 SW1116细胞中PLK1 mRNA及蛋白都呈现高表达 3种干扰片段转染SW1116细胞后,PLK1表达量均有不同程度下降,以片段1干扰效果最为明显.在迁移实验中,PLK1干扰组穿膜细胞数为（44±14）个,低于阴性对照组[（242±40）个]和空白对照组[（240±38）个] 在侵袭实验中,PLK1干扰组穿膜细胞数为（62±3）个,低于阴性对照组[（207±12）个]和空白对照组[（211±15）个] 上述差异均有统计学意义（P＜0.01）.结论 干扰PLK1能显著抑制肿瘤细胞的迁移、侵袭能力,提示PLK1可能在结直肠癌浸润和转移中具有一定作用.     

NDRG2基因对前列腺癌细胞株PC-3M侵袭转移能力的影响

目的 观察NDRG2基因对人前列腺癌细胞株PC-3M侵袭转移能力的影响.方法 以携带人NDRG2基因的腺病毒感染体外培养的PC-3M细胞株,采用Western blot及明胶酶谱实验检测NDRG2、MMP-2、MMP-9蛋白表达情况及相关酶活性的变化.平板克隆实验、细胞生长实验检测NDRG2对PC-3M增殖能力的影响.Transwell实验检测NDRG2对PC-3M细胞体外侵袭能力的影响.结果 Ad-NDRG2感染后,PC-3M细胞中NDRG2表达明显增加,而MMP-2和MMP-9表达水平及活性均降低.噻唑蓝(MTT)比色法(抑制率24 h为16.2%、48 h为24.4%、72 h为43.7%)及平板克隆实验(3组分别为56.3%、55.2%和36.7%)显示NDRG2对PC-3M细胞的生长有明显抑制作用.Transwell显示对照组及Ad-LacZ组穿入下室面的细胞数明显多于Ad-NDRG2组(3组分别为93.0、94.8和50.4).结论 NDRG2对人前列腺癌PC-3M细胞株侵袭能力有明显抑制作用,提示NDRG2可能在前列腺癌的侵袭及转移中发挥重要作用.

Abstract：

Objective To investigate the effect of NDRG2 gene expression on the cell migration and invasion of human prostate cancer cell line PC-3M.Methods Recombinant adenovirus vectors carrying human NDRG2 gene (Ad-NDRG2) were infected into prostate cancer cell line PC-3M.The protein expression and enzymatic activities of NDRG2,MMP-2 and MMP-9 were determined by Western blotting and gelatin zymography respectively.Methylthiazol tetrazolium (MTT) assay and plate colony formation were used to determine the effect of proliferation of PC-3M cells.Invasion of PC-3M cells was measured by Transwell chamber assay.Results After infection by Ad-NDRG2,it had been verified that the protein expression of NDRG2 in PC-3M cells was obviously increased,and the expression and enzymatic activities of MMP-2 and MMP-9 were reduced.The MTT assay (inhibition ratio: 24 h,16.2%,48 h,24.4%,and 72 h,43.7% respectively) and plate colony formation (56.3%,55.2% and 36.7% in control group,Ad-LacZ group and Ad-NDGR2 group respectively) revealed that NDRG2 could significantly inhibit the growth and proliferation of PC-3M cells.The number of PC-3M cells that invaded the lower chamber in the Ad-NDRG2 group was significantly decreased as compared with the control group and the Ad-LacZ group (93.0,94.8 and 50.4 respectively).Conclusion NDRG2 gene can significantly inhibit invasion of the PC-3M cells,which may paly an important role in metastasis of prostate cancer.     

miR-152对前列腺癌细胞株迁移和侵袭能力的影响

目的研究miR-152在前列腺癌、前列腺正常组织中的表达情况及其在前列腺癌细胞系中的作用。方法采用TaqMan荧光定量RT—PCR方法检测8例前列腺癌和8例前列腺正常组织的样本中miR-152的表达水平。运用Transwell细胞迁移实验及侵袭实验评估miR一152对前列腺细胞系PC-3和DU145细胞功能的影响。结果与正常前列腺组织相比,miR-152在前列腺癌组织中的表达水平显著下调（P〈0．05）。体外实验中上调miR152的表达可以显著降低前列腺癌细胞的迁移和侵袭能力（P〈O．05）。结论miR-152在前列腺癌中可作为一种肿瘤抑制因子,影响前列腺癌细胞的迁移和侵袭能力。     

趋化素样因子超家族成员5对前列腺癌侵袭行为的影响

目的 探讨趋化素样因子超家族成员5(CMTM5)对前列腺癌细胞的作用及机制.方法 利用划痕实验观察CMTM5对前列腺癌、DU145细胞迁移的影响;蛋白质印迹法检测PI3 K-AKT信号通路相关蛋白的表达;建立18只裸鼠皮下前列腺癌移植瘤模型,实验组肿瘤局部注射CMTM5腺病毒,检测CMTM5过表达对裸鼠前列腺肿瘤生长的影响,应用免疫组织化学方法检测裸鼠肿瘤组织中Ki-67的表达. 结果 过表达CMTM5抑制前列腺癌DU145细胞迁移,减少了PI3K-AKT信号通路中的关键分子pAKT 、NF-kB等蛋白的表达.裸鼠体内实验结果显示,CMTM5组肿瘤体积及质量分别为(573.39±175.24)mm3及(0.55±0.11)g,对照组分别为(1482.50±327.86) mm3及(1.31±0.29)g,2组比较差异均有统计学意义(P＜0.05).CMTM5组肿瘤组织Ki-67表达明显低于对照组. 结论 过表达CMTM5抑制前列腺肿瘤的增殖及迁移能力,其机制与PI3K-AKT信号通路有关.     

Small interference RNA-mediated silencing of prostate stem cell antigen attenuates growth,reduces migration and invasion of human prostate cancer PC-3M cells

ObjectivesProstate stem cell antigen (PSCA), a glycosylphosphatidylinositol (GPI)-anchored cell surface glycoprotein, is highly expressed in both local and metastatic prostate cancer (CaP). Elevated PSCA expression has been shown to correlate with malignant phenotype and clinical progression. The purpose of the current study is to investigate the therapeutic potential of small interference RNA (siRNA) targeting PSCA on human CaP cells.Materials and methodsA set of two siRNAs directed different regions of human PSCA (siRNA-PSCA) were designed and transfected into a human CaP PC-3M cell line. The silencing effect was screened by RT-PCR and Western blotting. The biological effects of siRNA-PSCA on PC-3M cells were investigated by examining the cell proliferation through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell cycle distribution through flow cytometry, and migration and invasion potencies through transwell invasion assay upon the PSCA silencing.ResultsPC-3M cells had positive PSCA expression on immunocytochemical assay. PSCA expression was depleted at 48 hours after transfection with siRNA-PSCA. Silencing of PSCA significantly suppressed cell proliferation. Cell cycle assay showed that the anti-proliferation effect of siRNA-PSCA was mediated by arresting cells in the G₀/G₁ phase rather than apoptosis. Furthermore, PSCA knockdown resulted in a marked decrease of cell migration and invasion capabilities in PC-3M cells.ConclusionsThe present study provides the first evidence that silencing PSCA using siRNA can inhibit the proliferation and invasiveness properties of human CaP cells, which may provide a promising therapeutic strategy for CaP and open a novel avenue toward the investigation of the role of PSCA overexpression in cancers.     

RhoA基因沉默对肝癌HepG2细胞增殖和迁移能力的影响

Objective To construct a RhoA-siRNA expression vector and determine its role on the malig-nant behavior of HepG2 cells.Methods A RhoA-siRNA DNA fragment was synthesized and cloned into the expression vector of pGenesil-1.The constructed Rhon-siRNA DNA plasmid was stably transfected into HerG2 cells by lipofectamine,and then HepG2 cells were divided into the HepG2/RhoA-siRNA group (HepG2 cells were transfected with pGenesil-1-RhoA-siRNA),HepG2/control group(HepG2 cells were transfected with control plasmid) and HepG2 group (without plasmid transfection).The inbibitory effect of RhoA-siRNA on RhoA protein expression was shown by Western blot.The proliferation,migration,growth potentiality and cell cycle of transfected HepG2 cells were evaluated by MTT assay,wounded healing,the plate cloning formation test and flow cytometry,respectively.All data were analyzed by one-way analysis of variance (ANOVA) and chi-square test.Results The expression of RhoA protein in the HepG2/RhoA-siRNA group was,significantly decreased compared with that in the other two groups (F=178.19,P<0.05).Scratched cells were healed within 48 hours in the HepG2/control group and HepG2 group,but not in the HepG2/RhoA-siRNA group.The clone formation rates in the HepG2/RhoA-siRNA group,HepG2 group and HepG2/control group were 39%±3%,67%±5%and 70%±6%,respectively,with a significant difference among the three groups(χ2=33.34,38.69,P<0.05).Flow cytometry showed that the number of cells transfected with RhoA-siRNA was highest in the G0/G1 phase and lowest in the S phase(F=70.46,76.57.P<0.05).Conclusion The RhoA-siRNA expression vector can effectively suppress the proliferation and migration of HepG2 cells,which may provide a novel gene therapy for hepatocellular carcinoma.     

RhoA基因沉默对肝癌HepG2细胞增殖和迁移能力的影响

目的 构建RhoA-siRNA表达载体,研究其对肝癌HepG2细胞肿瘤生物学行为的影响.方法 利用pGenesil-1 质粒构建RhoA-siRNA表达载体,以脂质体法转染至肝癌HepG2细胞中建立稳定细胞系,并分为3组.转染 pGenesil-1-RhoA-siRNA 载体者为HepG2/RhoA-siRNA组,转染随机对照载体者为HepG2/control组,未转染的肝癌HepG2细胞作为HepG2组.Western blot检测RhoA-siRNA对其蛋白表达的抑制情况.分别采用MTT法、细胞划痕损伤和平板克隆形成实验检测转染细胞的增殖、迁移和生长潜能,流式细胞仪检测细胞周期变化.采用单凶素方差分析、x2检验比较各组差异.结果 3组细胞蛋白表达水平比较,HepG2/RhoA-siRNA组RhoA蛋白的表达明显下调(F=178.19,P<0.05).HepG2/control组和HepG2组细胞划痕损伤在48 h内愈合,而HepG2/RhoA-siRNA组则不能愈合.HepG2/RhoA-siRNA组克隆形成率低于HepG2组和HepG2/control组,分别为39%±3%、67%±5%、70%±6%,其差异有统计学意义(χ2=33.34,38.69,P<0.05).RhoA基因沉默显著抑制肝癌HepG2细胞的增殖,细胞周期中G0/G1期细胞数量增多而S期细胞数量减少(F=70.46,76.57,P<0.05).结论 RhoA-siRNA表达载体能抑制肝癌HepG2细胞的增殖和迁移,可为肝癌的基因治疗提供新的方法.     

RhoA基因沉默对肝癌HepG2细胞增殖和迁移能力的影响

Objective To construct a RhoA-siRNA expression vector and determine its role on the malig-nant behavior of HepG2 cells.Methods A RhoA-siRNA DNA fragment was synthesized and cloned into the expression vector of pGenesil-1.The constructed Rhon-siRNA DNA plasmid was stably transfected into HerG2 cells by lipofectamine,and then HepG2 cells were divided into the HepG2/RhoA-siRNA group (HepG2 cells were transfected with pGenesil-1-RhoA-siRNA),HepG2/control group(HepG2 cells were transfected with control plasmid) and HepG2 group (without plasmid transfection).The inbibitory effect of RhoA-siRNA on RhoA protein expression was shown by Western blot.The proliferation,migration,growth potentiality and cell cycle of transfected HepG2 cells were evaluated by MTT assay,wounded healing,the plate cloning formation test and flow cytometry,respectively.All data were analyzed by one-way analysis of variance (ANOVA) and chi-square test.Results The expression of RhoA protein in the HepG2/RhoA-siRNA group was,significantly decreased compared with that in the other two groups (F=178.19,P<0.05).Scratched cells were healed within 48 hours in the HepG2/control group and HepG2 group,but not in the HepG2/RhoA-siRNA group.The clone formation rates in the HepG2/RhoA-siRNA group,HepG2 group and HepG2/control group were 39%±3%,67%±5%and 70%±6%,respectively,with a significant difference among the three groups(χ2=33.34,38.69,P<0.05).Flow cytometry showed that the number of cells transfected with RhoA-siRNA was highest in the G0/G1 phase and lowest in the S phase(F=70.46,76.57.P<0.05).Conclusion The RhoA-siRNA expression vector can effectively suppress the proliferation and migration of HepG2 cells,which may provide a novel gene therapy for hepatocellular carcinoma.     

TPM4对人胰腺癌细胞侵袭和迁移的影响

目的 探讨原肌球蛋白4(TPM4)对人胰腺癌细胞侵袭和迁移的影响。方法 采用qRT-PCR检测人正常胰腺导管上皮细胞(HPDE)和人胰腺癌细胞系(ASPC-1、BXPC-3、MIA PaCa-2、PANC-1、SW1990)中TPM4 RNA表达量。采用siRNA或过表达慢病毒感染胰腺癌细胞(MIA PaCa-2、PANC-1),通过qRT-PCR、Western blotting检测各组细胞TPM4的表达,划痕实验及Transwell实验检测各组胰腺癌细胞侵袭和迁移能力。结果 GEPIA数据库中胰腺癌组织TPM4表达明显高于正常胰腺组织(P<0.05),qRT-PCR分析TPM4 RNA在胰腺癌细胞系中表达均高于HPDE(P<0.0001)。与对照组相比,在MIA Paca-2和PANC-1中过表达TPM4促进了胰腺癌细胞侵袭迁移能力(均P<0.01),而敲低TPM4胰腺癌细胞侵袭迁移能力则明显减弱(均P<0.01)。结论 TPM4在胰腺癌细胞中呈现高水平表达,可能在胰腺癌中发挥着癌基因的作用,其并可促进胰腺癌细胞迁移及侵袭能力。     

NK4基因转染对前列腺癌DU145细胞增殖、迁移、侵袭和凋亡的影响

研究HGF拮抗剂NK4在前列腺癌中的作用。将包含NK4cDNA的表达载体pBudCE4．1-EGFP-NK4转染到DU145细胞中。体外实验检测白分泌的NK4对肿瘤细胞增殖、迁移、转移及凋亡的影响。体内实验裸鼠分为三组,分别皮下种植DUl45、空质粒转染的Dul45和NK4转染的DUl45细胞,检测皮下肿瘤的大小、细胞凋亡及细胞增殖情况。体外实验结果显示转染NK4的DUl45细胞可以分泌NK4蛋白。自分泌的NK4抑制HGF诱导的肿瘤细胞增殖、迁移及转移,促进凋亡（P〈0．01）。NK4可以调节HGF受体c－Met及其下游ERKl与Akt1／2蛋白的活性。体内实验显示,NK4转染DUl45细胞组的肿瘤生长及细胞增殖受到抑制,同时肿瘤细胞凋亡增加。本实验显示包含NK4cDNA的表达载体转染前列腺癌细胞可以有效地调节肿瘤细胞的增殖、迁移、侵袭及凋亡。NK4作用于HGF／c－Met可以作为前列腺癌治疗的一个有效靶点。