LYRM1基因过表达对3T3-L1脂肪细胞线粒体代谢的影响

目的观察LYRM1基因过表达对3T3-L1脂肪细胞线粒体代谢的影响。方法体外培养3T3-L1前体脂肪细胞,分别构建LYRM1基因过表达细胞株(LYRM1-pcDNA3.1/myc-His B)和空载对照细胞株(pcDNA3.1/myc-His B);实时定量RT-PCR验证转染情况;RT-PCR检测诱导分化成熟后的线粒体代谢酶HKI,ACC,CS,CPT1和Cyc-C基因表达水平。结果 RT-PCR结果证实LYRM1质粒转染成功;与空载对照组比较,LYRM1过表达组HKI,ACC,CS,CPT1和Cyc-C mRNA表达水平均显著降低(P均<0.05)。结论 LYRM1基因过表达下调了3T3-L1脂肪细胞中线粒体的代谢关键酶基因表达水平,可能参与调节脂肪细胞线粒体的代谢。     

膜联蛋白A4对肝癌细胞株MHCC97H黏附能力及黏附相关基因表达的影响

目的探讨人膜联蛋白A4(AnnexinA4,ANXA4)对肝癌细胞株MHCC97H黏附能力的影响及其相关机制。方法化学合成针对ANXA4序列特异性小干扰RNA,用脂质体LipofectamineTM2000转染人肝癌细胞株MHCC97H;RT-PCR和western blot检测RNA干扰后ANXA4 mRNA和蛋白质的表达水平;黏附实验检测ANXA4表达下降对细胞黏附能力的影响;荧光定量RT-PCR检测RNA干扰后黏附相关基因表达的变化。结果荧光定量RT-PCR及western blot结果表明,经siRNA干扰后,ANXA4 mRNA和蛋白质表达水平明显降低;细胞黏附实验结果显示,与空白对照组的0.293±0.019和Mock组的0.305±0.018相比,实验组为0.234±0.013,差异有统计学意义(P<0.05)。荧光定量RT-PCR结果显示,与阴性对照组相比,实验组ITGA5、ITGB1、E-cadherin mRNA表达水平下调,ICAM1、CD44、OPN mRNA表达水平上调,差异均有统计学意义(P均<0.05)。结论 ANXA4可能通过调节黏附相关基因的表达水平影响细胞的黏附能力,参与肝癌的发生和发展进程。     

LNK基因沉默和过表达对THP-1细胞STAT3表达影响的研究

目的:探讨LNK基因沉默和过表达对人单核细胞白血病细胞(THP-1)STAT3基因表达的影响。方法:培养THP-1细胞;以慢病毒为载体介导SiRNA、LNK(SH2B3)稳定沉默及过表达LNK基因;转染72 h后于倒置荧光显微镜下观察细胞的GFP表达量;应用流式细胞术检测慢病毒感染效率;RT-PCR验证LNK沉默及过表达效应,检测STAT3 mRNA表达量;Western blot检测LNK、STAT3蛋白水平。结果:慢病毒感染72 h THP-1细胞GFP表达量达85%以上,细胞感染效率在99%以上。LNK沉默组LNK及STAT3 mRNA表达量明显低于对照组,而LNK过表达组的LNK和STAT3 mRNA表达水平明显高于对照组(P 0.05)。LNK沉默组LNK、STAT3蛋白水平明显低于对照组,而在LNK过表达组明显高于对照组(P 0.05)。结论:成功建立了LNK基因沉默及过表达的THP-1细胞株。沉默LNK基因导致STAT3表达下调;过表达LNK基因,使STAT3表达上调;此结果提示LNK可能参与了STAT3的调控。     

葡萄糖对INS-1细胞线粒体谷氨酸转运体1基因表达的影响

目的观察葡萄糖对小鼠胰腺β瘤细胞INS-1线粒体谷氨酸转运体1(MGC1)基因表达的影响。方法分别采用半定量RT-PCR和Western blot,检测葡萄糖对INS-1细胞MGC1基因mRNA和蛋白表达的影响。结果葡萄糖浓度的提高对INS-1细胞MGC1基因mRNA和蛋白的表达有明显的增强作用(P<0.05)。结论高浓度葡萄糖能增强INS-1细胞MGC1基因表达。     

慢病毒介导KSHV vIL-6基因稳定表达的血管内皮细胞株的建立

目的建立慢病毒介导且稳定表达病毒白细胞介素6(viral interleukin-6,vIL-6)蛋白的血管内皮细胞株。方法以质粒pvIL-6-Flag为模板,PCR扩增vIL-6基因,克隆至慢病毒载体p3DLV,经酶切和测序鉴定后,将重组质粒与慢病毒辅助包装元件质粒共转染293T细胞,构建含vIL-6基因的重组慢病毒。重组慢病毒感染血管内皮细胞EA.hy926,经潮霉素筛选,获得稳定表达细胞株,western blot鉴定vIL-6蛋白的表达。结果酶切鉴定和基因测序证实长度为615 bp的vIL-6基因成功克隆至慢病毒表达载体;重组慢病毒经包装、纯化后测得滴度为1.0×107 TU/mL。重组慢病毒感染EA.hy926细胞,经潮霉素筛选,形成生长形态良好的单克隆细胞株EA.hy926-vIL-6。western blot鉴定该细胞株可稳定表达vIL-6蛋白。结论建立了稳定表达vIL-6的EA.hy926细胞株,为进一步研究vIL-6的生物学功能及致病机制提供了细胞模型。     

Bmi-1 shRNA慢病毒表达载体的构建及稳转U266细胞株的建立

本研究构建Bmi-1基因的shRNA慢病毒表达载体,建立U266-li稳定细胞株,为下一步Bmi-1的功能研究及应用RNAi技术治疗打下基础。设计、合成1对针对Bmi-1 mRNA的shRNA序列,退火后连接到pLVTHM干扰载体上,与psPAX2、PMD2G共转染HEK 293T细胞,包装产生慢病毒颗粒并测定病毒滴度,感染U266细胞,建立稳定细胞株;应用实时PCR和Western blot技术分别检测U266稳定细胞中Bmi-1及P14 mRNA和蛋白水平的表达,并与对照组进行比较。结果表明,成功构建了针对Bmi-1基因的shRNA慢病毒表达载体,病毒滴度为5×107 TU/ml;建立稳定转染的U266细胞株。有效干扰验证显示,shBmi-1能明显降低Bmi-1的mRNA及蛋白水平,而P14的mRNA及蛋白水平则都上调,其差异有统计学意义(P<0.05)。结论:成功构建Bmi-1基因的shRNA慢病毒表达载体,建立稳定干扰Bmi-1表达的U266细胞株。     

IK6过表达慢病毒载体的构建及其在THP1细胞系中的表达和生物学特征

本研究旨在构建人IK6基因过表达的慢病毒载体,观察其在THP1细胞株中的表达及对其生物学特征的影响,为研究该基因在白血病中的作用提供基础.采用RT-PCR方法获得目的基因,并与线性化慢病毒载体pGC-FU重组构建慢病毒载体pGC-FU-IK6-GFP,通过菌落PCR鉴定、测序比对分析,确定克隆的正确性.使用Lipofectamine 2000,将慢病毒载体pGC-FU-IK6-GFP转染293T细胞,测定病毒滴度后转染THP1细胞株,用流式细胞仪检测转染效率,RT-PCR检测靶细胞内IK6 mRNA表达,Western blot检测IK6-GFP融合蛋白,进一步观察THP1生物学特征的改变.结果表明,利用RT-PCR方法获得IK6目的基因并连接到线性化的慢病毒载体上,成功构建具有Amp抗性的pGC-FU-IK6-GFP质粒并转染293T细胞,获得滴度为2.0×109TU/ml的病毒.用目的质粒转染THP1细胞,转染效率可达90％.在靶细胞中检测到IK6 mRNA表达和IK6-GFP融合蛋白表达.IK6能够促进靶细胞克隆形成并且具有抑制凋亡的作用,而对细胞周期无显著影响.结论：成功构建IK6过表达的慢病毒载体,使IK6在THP1细胞株中稳定表达.对靶细胞生物学研究发现IK6极为可能干扰了正常Ikaros蛋白的抑癌作用,并存在潜在的抗凋亡作用,进而在一定程度上促进白血病细胞生长,但对细胞周期无明显影响.该研究为进一步探讨IK6在急性髓系白血病的作用奠定了基础.     

HSF1在热休克反应中对KLF4基因表达的影响

[目的]观察热休克因子1(HSF1)在热休克反应中对Kruppel 样因子4(KLF4)基因表达的影响;采用生物信息学方法初步探讨KLF4在热休克反应中调控的下游基因.[方法]采用HSF1基因敲除小鼠热休克模型,抽提HSF1基因敲除小鼠(HSF1^-/-)和野生型小鼠(HSF1^ / )心肌及肺组织的总RNA进行RT-PCR和Northern blot实验,观察KLF4mRNA表达的情况.用热休克处理和HSF1过表达的小鼠RAW264.7巨噬细胞,抽提总RNA进行RT-PCR实验,观察KLF4mRNA表达的情况;用TESS分析启动子含有KLF4结合位点的下游基因.[结果]热休克处理后,HSF1^ / 小鼠组织中KLF4mRNA的水平明显增加,HSF1^-/-小鼠组织中KLF4 mRNA水平的增加明显低于HSF1^ / 小鼠.小鼠RAW264.7巨噬细胞受热刺激后,KLF4mRNA的水平明显增加;在HSF1过表达细胞中KLF4的表达也明显增高.经TESS软件分析发现6个启动子区含有KLF4结合位点的下游基因.[结论]HSF1诱导KLF4基因在热休克反应中呈现高表达.     

siRNA介导入结肠癌细胞诱骗受体3基因表达抑制的研究

目的：研究siRNA对结肠癌细胞株SW480DcR3基因表达的影响。方法：化学合成靶向DcR3基因的4组siRNA．用脂质体转染试剂转染结肠癌细胞株SW480,应用噻唑兰（MTT）法检测siRNA对细胞生长的作用,RT—PCR和Westem blot方法检测DcR3基因mRNA水平和蛋白表达量的变化。结果：与对照组相比,MTT试验结果显示各组siRNA对细胞增殖均有明显的抑制效应．但此抑制效应随时间延长而减弱。Western blot结果显示DcR3蛋白表达量明显降低。RT-PCR显示．转染后细胞内DcR3mRNA的表达量与空白对照组相比降低至24％。结论：DcR3siRNA通过特异、高效地沉默结肠癌细胞DcR3mRNA的表达,发挥抑制结肠癌细胞生长的作用。     

蚕丝支架与3T3-L1前脂肪细胞生物相容性的体外实验(英文)

背景:蚕丝是天然制品,其力学性能及生物相容性优于传统人工合成的可降解高分子材料,在医疗领域中已获得了广泛的应用而受到关注.目的:观察蚕丝对3T3-L1前脂肪细胞吸附作用及蚕丝对3T3-L1前脂肪细胞形态和功能的影响.方法:取原料蚕丝和用胰酶消化后的蚕丝任意缠绕成网状立体构型纤维条索,架空固定在自制的不锈钢支架上,支架网孔70～200 μm,厚200-300 μm,孔隙率为20%.消化后的蚕丝三维支架放入24孔培养板中,将浓度为6×10L~(-1)的3T3-L1前脂肪细胞悬液每孔滴入3滴,每孔细胞数量为1×10~7个.悬空孵育4 h,待细胞充分吸附在支架上后加入培养液,令细胞完全浸没,隔两三天换半液,培养1～4周.结果与结论:①倒置显微镜观察3T3-L1前脂肪细胞-蚕丝复合物可见细胞伸出细长的突起沿着蚕丝不断向前迁移延伸,细胞首尾相互融合,渐渐连成一片分布于蚕丝网眼内.②扫描电镜观察3T3-L1前脂肪细胞-蚕丝复合物可见细胞与支架紧密贴附,适度伸展,并有基质分泌.提示蚕丝对3T3-L1前脂肪细胞具有良好的吸附作用,并能维持3T3-L1前脂肪细胞正常形态和功能.     

shRNA沉默IRS-1基因对鼠前脂肪细胞分化和PPARγ表达的影响

目的观察胰岛素受体底物-1（IRS-1）基因表达受阻对小鼠3T3-L1前脂肪细胞分化和脂质过氧化物酶体增殖物激活受体γ（PPARγ）表达的影响。方法针对小鼠IRS-1基因开放阅读框上的2个区域合成短发夹核糖核酸[shRNA（M和MH）],以pGenesil-1vector为载体构建带绿色荧光蛋白（GFP）靶标的shRNA质粒。以脂质体LipofectaminTM2000介导转染3T3-L1前脂肪细胞,并设阴性对照（HK）载体转染组。细胞转染后,G418筛选稳定表达的阳性克隆并通过免疫印迹法鉴定。应用0.5mmol/L3-异丁基-1-甲基黄嘌呤（IBMX）、10-6mol/L地塞米松（DEX）和5μg/mL胰岛素（Ins）分别诱导小鼠3T3-L1前脂肪细胞空白对照组、HK载体转染组、M和MH转染组分化,在诱导分化的不同时间点收集细胞。用免疫印迹法检测PPARγ的表达。脂肪细胞内脂滴用油红O染色观察。结果与空白对照组、HK组和转染M组IRS-1shRNA质粒的3T3-L1前脂肪细胞相比,转染MH组IRS-1shRNA质粒的3T3-L1前脂肪细胞IRS-1蛋白表达水平降低70%以上;而MH转染组细胞PPARγ蛋白的表达与前3组3T3-L1前脂肪细胞比较无改变。前3组3T3-L1前脂肪细胞诱导分化为成熟脂肪细胞成功,PPARγ蛋白的表达在脂肪细胞分化成熟过程中逐渐增高。油红O染色显示,随着脂肪细胞的分化成熟,桔红色脂滴逐渐增多;而转染MH组IRS-1shRNA质粒的3T3-L1前脂肪细胞经诱导,油红O染色桔红色脂滴显着减少,直到分化的第8天,才见少许桔红色脂滴,且PPARγ蛋白的表达无变化。结论 IRS-1基因沉默后可抑制3T3-L1前脂肪细胞的分化,并在分化过程中抑制PPARγ的表达,说明IRS-1通过调节PPARγ的表达或与PPARγ共同作用在3T3-L1前脂肪细胞的分化中发挥决定性作用。     

对腰大池持续脑脊液引流穿刺点的MRI分析

目的为探讨持续腰大池脑脊液引流（Continued lumbar cerebrospinal fluid drainage,CLCFD）穿刺点的解剖学依据和提高引流效果。方法选择64例行腰椎MRI检查的成年人,正中矢状面扫描进行穿刺点解剖学测量分析,即从L3-4和L4-5椎间盘的中点做一与椎管垂直的直线,命名为H1和H2线;该线与椎管前后径交点的中点即为两椎间隙的测量点（A点为L3-4,B点为L4-5）。分别测量A、B点到脊髓圆锥和硬膜囊终点的直线距离,H1和H2线与皮肤的交点S1和S1到蛛网膜下腔的距离。结果S1和S1到蛛网膜下腔的距离分别为（51．3±7．4）mm,（53．3±8．4）mm。A、B点到脊髓圆锥的距离分别为（90．1±16．4）mm,（121．9±17．3）mm;A、B点到硬膜囊终点的直线距离分别为（91．1±14．4）mm,（61．6±13．0）mm。比较A、B点到脊髓圆锥的距离及B点到脊髓圆锥和硬膜囊终点直线距离的差异有统计学意义。结论首选L4-5间隙穿刺头向置管;若选择L3-4间隙穿刺,头向和尾向置管都可行,但头向置管有利于引流。     

Prostate tumor microenvironment alters immune cells and prevents long-term survival in an orthotopic mouse model following flt3-ligand/CD40-ligand immunotherapy

A novel orthotopic metastatic model of mouse prostate cancer was developed using MHC-negative TRAMP-C1P3 (transgenic adenocarcinoma of mouse prostate) cells derived by serial passage of the parental TRAMP-C1 line in mouse prostate glands. TRAMP-C1P3 cells grew efficiently in mouse prostate glands and reproducibly metastasized to draining lymph nodes. Using this model, we show that Fms-like tyrosine kinase-3 ligand (flt3-L) dramatically inhibited growth of preexisting orthotopic TRAMP-C1P3 tumors and the development of metastatic disease. Mice remained in remission for several months following termination of flt3-L treatment but eventually relapsed and died of progressive disease. flt3-ligand treatment induced a pronounced mixed inflammatory cell infiltrate that consisted of CD8alpha-CD4- dendritic cells (CD11c+), macrophages, granulocytes (Gr-1+) and to a lesser extent T cells (CD4+ and CD8+). Dendritic cells isolated from TRAMP-C1P3 tumors were phenotypically immature (CD11c+ B7.2-I-A-CD40-), and this phenotype was also predominant in peripheral organs of mice treated with flt3-L alone or in combination with the DC maturation factor, CD40-L. Diminished expression of TCR-beta, CD3-epsilon, and CD3-zeta was also observed on intratumoral T cells, although these signaling proteins were reexpressed following in vitro culture with IL-2. The TCR/CD3 complex remained intact on peripheral T cells except in mice treated with flt3-L where CD3-zeta loss was observed. In contrast to alphabeta-T cells, tumor-infiltrating gammadelta-T cells maintained expression of their antigen receptors but not CD3epsilon. Thus, TRAMP-C1P3 tumors quickly establish a microenvironment that profoundly diminishes expression of molecules critical for normal dendritic cell and T cell function, thus limiting the efficacy of flt3-L and CD40-L immunotherapy. Overall, these data suggest that long-term cures of established MHC-negative tumors may not be achieved until therapeutic interventions are engineered to overcome this immunosuppressive microenvironment.     

Alkaline phosphatase is involved in the control of adipogenesis in the murine preadipocyte cell line, 3T3-L1

OBJECTIVE: As alkaline phosphatase may play a role in cell differentiation, our aim was to study the possible role of this enzyme in the differentiation of preadipocytes (3T3-L1 cells) into adipocytes. RESEARCH METHODS AND PROCEDURES: 3T3-L1 cells were grown in medium containing insulin, dexamethasone and IBMX to induce adipogenesis. Adipogenesis was measured using the triglyceride-specific dye, oil red O at 0, 3, 7 and 11 days after initiation of adipogenesis in the presence or absence of the alkaline phosphatase inhibitors, levamisole, histidine and Phe-Gly-Gly. Intracellular localisation of the enzyme was detected using ELF-phosphatase, a fluorescent substrate and alkaline phosphatase gene expression was assessed using RT-PCR. RESULTS: Alkaline phosphatase activity was detected in untransformed cells (1.91+/-0.62 mU/mg protein) and activity increased 11.5+/-1.4-fold after 11 days treatment with transformation medium and 5.3+/-0.3-fold in transformation medium containing levamisole (p<0.05). Triglyceride content of cells increased 3.1+/-0.2-fold after 11 days treatment with transformation medium and 2.1+/-0.3-fold in the presence of levamisole (p<0.005). Histidine inhibited adipogenesis and alkaline phosphatase to a greater extent than did levamisole, but Phe-Gly-Gly had no effect on these variables. Alkaline phosphatase was localised around the lipid droplets of the cells. Gene expression of alkaline phosphatase increased during adipogenesis. DISCUSSION: This study demonstrates that tissue-nonspecific alkaline phosphatase is present in 3T3-L1 cells and that it may play a role in the control of adipogenesis.     

人乳头瘤病毒16型L1基因真核表达载体的构建及表达

目的从感染人乳头瘤病毒（HPV）的新鲜宫颈癌组织中扩增HPV16型晚期蛋白L1基因全长，构建表达载体，并验证目的蛋白的表达。方法根据基因全长设计一对特异引物，用PCR的方法从宫颈癌组织DNA，中获得L1基因，以pMD18T为克隆载体，构建重组质粒进行亚克隆，再以pcDNA3．1（＋）为载体构建表达质粒，转染至人肝细胞系H7702，提取蛋白，采用Western Blot方法检测HPV16-L1蛋白的表达。结果从长春地区宫颈癌临床标本克隆到的HPV16型L1蛋白编码序列与Genebank报道序列高度同源，其真核表达产物与特异性抗体能够特异性结合。结论成功构建重组表达质粒pcDNA3．1（＋）-HPV16-L1，为进一步研制HPV预防性疫苗创造基础条件。     

Recombinant human CD40 ligand stimulates B cell proliferation and immunoglobulin E secretion

Signaling through the cell surface molecule, CD40, is known to play an important role in the proliferation and differentiation of B lymphocytes. Using the thymoma cell line EL4, we recently identified and cloned a cDNA encoding a murine ligand for the CD40 molecule (mCD40-L) and showed that it has biological activity in vitro. A cDNA encoding a human homologue of the mCD40-L was isolated using crosshybridization techniques from an activated peripheral blood T cell library. The predicted amino acid sequence indicates that this human ligand for CD40 (hCD40-L) is a 261 amino acid type II membrane protein that exhibits 78% amino acid identity with its murine counterpart. Northern blot and FACS analyses suggest that the hCD40-L is restricted in its expression to T lymphocytes, and that it is most abundant on the CD4+ T cell subpopulation. Cells transfected with hCD40-L caused the proliferation of human tonsil B cells in the absence of costimuli and, in the presence of interleukin 4, induced immunoglobulin E secretion from purified human B cells. A comparison of the efficacy of the hCD40-L and mCD40-L in these assays is presented.     

Functional CD40 ligand is expressed by T cells in rheumatoid arthritis.

CD40 ligand (CD40-L), a member of the tumor necrosis family of transmembrane glycoproteins, is rapidly and transiently expressed on the surface of recently activated CD4+ T cells. Interactions between CD40-L and CD40 induce B cell immunoglobulin production as well as monocyte activation and dendritic cell differentiation. Since these features characterize rheumatoid arthritis (RA), the expression and function of CD40-L in RA was examined. Freshly isolated RA peripheral blood (PB) and synovial fluid (SF) T cells expressed CD40-L mRNA as well as low level cell surface CD40-L. An additional subset of CD4+ RA SF T cells upregulated cell surface CD40-L expression within 15 min of in vitro activation even in the presence of cycloheximide, but soluble CD40-L was not found in SF. CD40-L expressed by RA T cells was functional, since RA PB and SF T cells but not normal PB T cells stimulated CD40-L-dependent B cell immunoglobulin production and dendritic cell IL-12 expression in the absence of prolonged in vitro T cell activation. In view of the diverse proinflammatory effects of CD40-L, this molecule is likely to play a central role in the perpetuation of rheumatoid synovitis. Of importance, blockade of CD40-L may prove highly effective as a disease modifying therapy for RA.     

ANGPTL4基因在3T3-L1前脂肪细胞诱导分化过程中表达水平的变化

目的观察血管生成素样蛋白-4（Angiopoietin-like protein 4,ANGPTL4）基因在3T3-L1前脂肪细胞诱导分化过程中表达水平的变化,探讨ANGPTL4基因与脂肪细胞分化的关系。方法体外培养3T3-L1前脂肪细胞,在3T3-L1前脂肪细胞分化成熟过程中（0-10 d）,采用油红O染色,观察脂肪细胞分化及脂质积聚情况;采用Real-time PCR、逆转录PCR技术检测脂肪细胞分化标志基因（LEPTIN,ADIPONECTIN）和ANGPTL4 mRNA表达;采用Western blot检测ANGPTL4蛋白表达。结果随着脂肪细胞诱导分化逐步深入,油红O染色显示,细胞中脂滴形成逐渐增多,达到占全视野95%以上;同时瘦素（LEPTIN）和脂联素（ADIPONECTIN）基因在诱导后的第2天开始表达并逐步增多,均提示脂肪细胞分化逐步成熟。ANGPTL4 mRNA和蛋白在前脂肪细胞中低表达,分化第2-6天ANGPTL4基因表达显著上调,分化第8-10天ANGPTL4基因表达保持在较高水平并趋于稳定。结论在3T3-L1前脂肪细胞分化过程中ANGPTL4基因表达上调,ANGPTL4基因可能参与了脂肪细胞分化。     

LIF真核表达载体的构建及其在MSCs中的稳定表达

目的在人骨髓间充质干细胞内大量稳定的表达人白血病抑制因子（human Leukemia Inhibitory Factor,hLIF）。方法应用RT-PCR方法从人蜕膜组织中扩增出白血病抑制因子,并将其构建于pcDNA3．1真核表达载体上。利用脂质体进行基因骨髓间充质干细胞的转染。利用G418进行基因的稳定表达筛选。通过RT-PCR和Westernblot方法进行基因表达的检测。结果成功扩增出人白血病抑制因子基因,成功构建hLIF-pcDNA3．1真核表达载体。转染LIF-pcDNA3．1的骨髓间充质干细胞经G418筛选,存活十代以上。经RT-PCR及Westernblot方法检测发现转染LIF-pcDNA3．1的骨髓间充质干细胞表达白血病抑制因子明显高于未转染的骨髓间充质干细胞。结论成功构建LIF-pcDNA3．1的真核表达载体并使其在骨髓间充质干细胞得到稳定表达。