靶向Ccndl基因的siRNA对大鼠晶状体上皮细胞增殖的抑制作用

目的 探讨靶向Cendl基因小干扰RNA(siRNA)对培养的大鼠晶状体上皮细胞(LECs)Ccndl mRNA和周期蛋白D1表达及细胞增殖的抑制作用.方法 合成3条靶向大鼠Ccndl基因的siRNA(si-Ccndl-1,si-Ccndl-2,si-Ccndl一3),LipofectamineTM2000转染体外培养的大鼠LECs,应用RT-PCR和Western blot方法分别检测Ccndl mRNA和周期蛋白D1表达变化,筛选最有效siRNA序列.MTT法连续测定转染最有效siRNA序列后24、48、72、96和120 h细胞增殖情况.结果 与对照组相比,3条siRNA均能不同程度地抑制Ccndl mRNA和周期蛋白D1的表达,以si-Ccndl-3最为有效(P<0.05).与对照组比较,si-Ccndl-3转染后24 h开始明显发挥对细胞增殖的抑制作用,并持续至120 h以后(P<0.05).结论 靶向Ccndl的siRNA能有效降低大鼠LECsCcndl mRNA和周期蛋白D1蛋白表达,抑制LECs增殖.     

短发夹RNA对前列腺癌细胞中PIM-1基因沉默效应的研究

目的:探讨脂质体介导的短发夹RNA(shRNA)对前列腺癌细胞中PIM-1基因表达的影响.方法:构建3种针对PIM-1 mRNA不同靶点的shRNA重组质粒表达载体,并转染前列腺癌PC-3细胞.分别采用逆转录-聚合酶链反应(RT-PCR)及蛋白印迹法(Westernblot)检测转染后细胞中PIM-1 mRNA及蛋白的表达情况.结果:转染48 h后,3种PIM-1基因靶向性shRNA表达质粒均可抑制PC-3细胞中PIM-1 mRNA和蛋白的表达,其中靶向PIM-1 mRNA第707-725位和1080-1098位序列的重组质粒干扰效果更显著.结论:本研究所构建的PIM-1 shRNA表达质粒可在体外高效特异地抑制前列腺癌细胞中PIM-1的表达,为下一步研究奠定了基础.     

RNA干扰质粒抑制富含半胱氨酸蛋白61在大鼠血管平滑肌细胞中的表达

目的观察RNA干扰质粒对大鼠血管平滑肌细胞中富含半胱氨酸蛋白61(Cyr61)表达的抑制作用。方法构建Cyr61RNA干扰质粒(pCyr61-shRNA)转染大鼠血管平滑肌细胞,采用半定量反转录聚合酶链反应及Western blot检测Cyr61 mRNA和蛋白表达。结果测序证实成功构建Cyr61RNA干扰质粒;转染pCyr61-shRNA组mRNA及蛋白表达均明显降低,与正常对照组和转染无关序列对照组(p-HK)相比差异有统计学意义(P<0.01)。结论RNA干扰质粒抑制Cyr61在大鼠血管平滑肌细胞中的表达。     

抑制IGF2基因的siRNA表达载体对肝癌Huh-7细胞增殖的抑制作用

目的:研究抑制人胰岛素样生长因子2(IGF2)基因的siRNA表达载体对肝癌Huh-7细胞增殖的抑制作用。方法:将重组人甲胎蛋白(hAFP)和人端粒酶逆转录酶(hTERT)双启动子调控抑制IGF2基因的siRNA表达载体pGL3-h AFP-hTERT-siRNA3(简称siRNA3)转染Huh-7细胞和正常肝L-02细胞,另设阴性对照(载体p GL3-hAFP-hTERT)组和空白对照组。采用实时荧光定量聚合酶链反应法检测各组细胞转染48 h后IGF2 mRNA表达;酶标仪检测转染0、24、48、72 h后的细胞活性;流式细胞仪检测转染48h后细胞周期、细胞凋亡;Western blot法检测细胞IGF2、增殖细胞核抗原(PCNA)、细胞周期蛋白(Cyclin)E2、Cyclin D2、Cdc2、Bcl-2蛋白表达水平。结果:与阴性对照组和空白对照组比较,转染siRNA3的Huh-7细胞中IGF2 mRNA表达明显减弱,转染48、72 h后Huh-7细胞活性明显降低,G1期Huh-7细胞明显增加,S期Huh-7细胞明显减少;Huh-7细胞早期、晚期及总凋亡百分比均明显增加,IGF2、PCNA、Cyclin E2、Cyclin D2、Cdc2和Bcl-2蛋白表达均明显减弱,以上差异具有统计学意义(P<0.01或P<0.05)。转染siRNA3表达载体的L-02细胞上述指标均无明显变化(P>0.05)。结论:重组hAFP和hTERT双启动子调控针对IGF2基因的siRNA可特异性抑制Huh-7细胞IGF2表达及细胞增殖,其可能与下调IGF2 mRNA及蛋白表达,进而引起细胞增殖相关基因PCNA、细胞周期调控相关基因Cyclin E2、Cyclin D2、Cdc2及细胞凋亡调控相关基因Bcl-2蛋白表达下调有关。     

RNA干扰对SKOV3卵巢癌细胞株Skp2表达的抑制作用

目的:探讨RNA干扰(RNAi)技术对SKOV3卵巢癌细胞株Skp2表达的抑制作用.方法:设计合成针对Skp2的小干涉RNA(siRNA)并进行转染.实验分为空白对照组、转染组1、转染组2、转染对照组及阴性对照组.采用Real time PCR和Western blotting技术检测各组Skp2 mRNA和蛋白水平的变化,通过细胞计数试剂盒-8(CCK-8)法检测各组卵巢癌细胞增殖情况.结果:应用Skp2-siRNA干扰SKOV3卵巢癌细胞株后,转染组1、转染组2的Skp2mRNA和蛋白表达水平明显降低,细胞增殖明显受限,与空白对照组相比较,差异有统计学意义(P＜0.05),转染组1与转染组2之间差异无统计学意义(P＞0.05).转染组1与转染组2基因沉默效率分别为75.31%和76.86%,蛋白抑制率分别为62.10%和63.11%,细胞增殖抑制率分别为52.75%和53.06%.结论:RNAi技术能够抑制SKOV3卵巢癌细胞株Skp2的表达,从而抑制卵巢癌细胞的增殖.     

RNAi介导大鼠腺苷A1和A2A受体基因沉默对乙醛诱导的肝星状细胞活化增殖的影响

目的研究大鼠腺苷A1受体(A1R)和腺苷A2A(A2AR)受体的siRNA分别转染大鼠肝星状细胞(HSC)对乙醛诱导的HSC活化增殖的影响。方法采用乙醛诱导HSC-T6建立离体的大鼠酒精性肝纤维化HSC模型,设计并合成A1R和A2AR小干扰RNA(small interfering RNA,siRNA)序列,通过脂质体LipofectamineTM2000瞬时转染至HSC-T6细胞内,荧光倒置显微镜观察细胞的转染效率,用四甲基偶氮唑盐(MTT)法检测HSC-T6细胞增殖变化;利用Real-Time q PCR及Western blot法分别检测HSC-T6的A1R、A2AR、α-SMA、Collagen I mRNA及蛋白表达。结果将A1R和A2AR siRNA转染至HSC-T6细胞内,A1R和A2AR基因及蛋白的表达水平明显降低;同时α-SMA、Collagen I mRNA及蛋白表达水平亦明显降低;靶向封闭A1R或A2AR基因的表达可明显抑制HSC-T6细胞的活化增殖。结论靶向封闭A1R或A2AR基因的表达可明显抑制HSC-T6细胞的活化增殖,A1R和A2AR可能是潜在的酒精性肝纤维化的治疗靶点。     

长链非编码 RNA膀胱癌相关转录物 1对微小 RNA-503-5p的靶向关系及对结直肠癌细胞增殖和凋亡的影响

目的 研究长链非编码RNA(lncRNA)膀胱癌相关转录物1(BLACAT1)对微小RNA(miR)-503-5p的靶向关系及结直肠癌细胞增殖和凋亡的影响.方法 实时荧光定量PCR(qRT-PCR)检测结肠癌细胞株SW620,LOVO,HT29和人正常结肠黏膜上皮细胞株NCM460中BLACAT1和miR-503-5p的表达.在HT29细胞中转染si-BLACAT1、pcDNA-BLACAT1或miR-503-5p,噻唑蓝(MTT)检测细胞增殖,流式细胞术检测细胞凋亡,蛋白质印迹法(Western blotting)检测细胞核相关抗原Ki-67(Ki-67)、细胞周期蛋白D1(Cyclin D1)、活化的多聚ADP-核糖聚合酶(Cleaved PARP)和活化的含半胱氨酸的天冬氨酸蛋白水解酶-3(Cleaved caspase-3)的表达,starbase预测和双荧光素酶报告分析BLACAT1与miR-503-5p之间的靶向关系.si-BLACAT1和anti-miR-503-5p共转染,观察干扰miR-503-5p对沉默BLACAT1诱导的结直肠癌细胞增殖和凋亡的影响.结果 与NCM460细胞比较,SW620、LOVO和HT29中BLACAT1表达量明显增加[(4.93±0.58)、(5.66±0.53)、(6.17±0.66)比(1.03±0.22)],miR-503-5p表达量减少[(0.72±0.11)、(0.67±0.09)、(0.51±0.08)比(1.04±0.14)](P<0.05).沉默BLACAT1或转染miR-503-5p明显减少HT29细胞的细胞存活率、Ki-67、CyclinD1蛋白表达量,提高细胞凋亡率、Cleaved PARP和Cleaved caspase-3蛋白水平(P<0.05),过表达BLACAT1则反之.BLACAT1靶向miR-503-5p调控其表达.干扰miR-503-5p部分逆转沉默BLACAT1抑制结直肠癌细胞增殖、Ki-67、CyclinD1蛋白表达和诱导结直肠癌细胞凋亡、Cleaved PARP、Cleaved caspase-3蛋白表达的作用.结论 lncRNA BLA-CAT1在表达上调,沉默BLACAT1可通过靶向调控miR-503-5p的表达,来抑制结直肠癌细胞增殖,并诱导细胞凋亡.     

hTERT siRNA对HepG2细胞hTERT表达及端粒酶活性的影响

目的研究siRNA技术对肝癌HepG2细胞系hTERT mRNA表达及端粒酶的抑制作用,探讨siRNA抑制肿瘤细胞的作用及机理。方法体外转录合成hTERT siRNA并转染入HepG2细胞株中,应用RT-PCR观察HepG2细胞中hTERT mRNA表达改变,Western blot检测hTERT蛋白的表达,应用末端重复片断扩增-酶联免疫吸附(TRAP-ELISA)方法测定转染24h、48h后细胞端粒酶活性。结果hTERT siRNA转染后,对HepG2细胞hTERT mRNA和蛋白表达明显抑制。转染后24h端粒酶活性转染组均数为(0.167±0.019),与未转染组均数(0.823±0.027)、空质粒阴性对照组均数(0.826±0.031)比较,差异有统计学意义(P<0.05);48h端粒酶活性转染组均数为(0.153±0.017),与未转染组均数(0.816±0.022)、空质粒阴性对照组均数(0.809±0.028)比较,差异有统计学意义(P<0.05)。结论靶向hTERT的siRNA,能有效抑制肝癌细胞系HepG2 hTERT-mRNA和蛋白表达及端粒酶活性。     

FLT3靶向RNA干扰对HL-60细胞增殖凋亡及cyclinD1、cyclinA表达的影响

目的探讨FLT3靶向RNA干扰对HL-60细胞增殖、凋亡及cyclinD1、cyclinA表达的影响。方法体外构建FLT3靶向短发夹状干扰RNA(FLT3-shRNA),转染HL-60细胞,RT-PCR、流式细胞术(FCM)鉴定FLT3的表达;CCK-8法检测细胞增殖活力;FCM检测细胞周期;AnnexinV染色法检测凋亡;RT-PCR及Western blot检测cyclinD1,cyclinA在mRNA、蛋白水平的表达。结果FLT3-shRNA转染可下调FLT3的表达,它对FLT3 mRNA和蛋白的抑制率分别达(81.66±10.25)%,(76.76±11.23)%。FLT3表达下降后细胞增殖活力受到抑制,转染48h抑制率达(31.66±2.97)%,72h达(33.10±3.43)%;细胞生长曲线低平,缺乏指数增殖特征。与对照组比,转染48h细胞周期重新分布,G0/G1期(58.48±6.17)%的明显上升,S期(27.72±5.10)%下降;细胞早期凋亡率(8.95±0.88)%上升;细胞内cyclinD1的mRNA、蛋白表达下降,cyclinA的表达无明显改变。结论FLT3靶向RNA干扰通过下调cyclinD1表达有效地抑制细胞增殖,具有潜在的临床应用价值。     

RNAi介导MeCP2基因沉默对HSC-T6细胞活化增殖的影响

目的探讨RNA干扰介导甲基CpG结合蛋白2(methyl-CpG-binding protein 2,MeCP2)基因沉默对大鼠肝星状细胞HSC-T6细胞增殖的影响。方法根据MeCP2的碱基序列设计并合成小干扰RNA(small interfering RNA,siR-NA),通过脂质体LipofectamineTM2000转染到HSC-T6细胞内,荧光倒置显微镜观察细胞的转染效率,用四甲基偶氮唑盐(MTT)法检测HSC-T6细胞增殖变化;Quantitative Real-Time PCR检测α-SMA、MeCP2 mRNA的表达;流式细胞术检测HSC-T6细胞周期的变化;Western blot检测α-SMA、MeCP2蛋白的表达。结果将MeCP2-siRNA转染进HSC-T6细胞内,MeCP2基因及蛋白水平明显降低;同时α-SMAmRNA及α-SMA蛋白的表达水平亦明显降低;靶向封闭MeCP2基因的表达可明显抑制HSC-T6细胞的活化增殖。结论靶向封闭MeCP2的表达可明显抑制HSC-T6细胞活化增殖,MeCP2可能是潜在的肝纤维化治疗靶点。     

siRNA pool targeting different sites of human hepatitis B surface antigen efficiently inhibits HBV infection

The main objective was to determine whether a pool of small interfering RNAs (siRNAs) targeting different regions of hepatitis B virus surface antigen (HBsAg) efficiently inhibits hepatitis B virus (HBV) infection. siRNAs targeting different regions of HBsAg were transfected into HBV-producing HepG2.2.15 cells and at 72 h post-transfection, the culture medium was collected for ELISA to determine HBsAg, while total RNA was isolated from the cells for real-time PCR. Three siRNA sequences that efficiently inhibited HBV infection were converted into small hairpin RNAs (shRNAs) and then cloned into a single plasmid psiSTRIKE driven by a single U6 promoter. These shRNA expressing plasmids were tested for HBsAg gene silencing in HepG2.2.15 cells. A pool of siRNAs targeting HBsAg efficiently inhibited HBV replication and antigen expression when transfected into HepG2.2.15 cells, compared with the use of single siRNA. Similarly, the plasmid encoding three different shRNAs driven by a single U6 promoter was more effective in silencing HBsAg at DNA, mRNA and protein levels compared with the plasmid encoding single shRNA. No apoptotic change was observed in the cells when the plasmid was transfected at a dose of 0.5-2 microg/1 x 10(6) cells after complex formation with Lipofectamine LTX. Furthermore, transfection with siRNA or shRNA did not increase interferon-gamma (IFNs-gamma) release, suggesting no induction of IFN response. In conclusion, a pool of chemically synthesised siRNAs as well as the shRNA expression plasmid encoding multiple shRNAs targeting different regions of HBsAg showed high gene silencing in HepG2.2.15 cells.     

黄芩苷通过调控c-Myc介导的细胞周期抑制胆管癌细胞增殖

目的　探究黄芩苷（BC）对人胆管癌（CCA）细胞QBC939和RBE增殖的影响及其潜在机制。方法　取对数生长期QBC939和RBE细胞，不同浓度的BC处理细胞24 h后，采用CCK-8法分别检测BC对CCA细胞增殖及敲低原癌基因蛋白质（c-Myc）后对BC引起细胞增殖活性改变的影响；高倍显微镜观察BC对细胞生长状态的影响；流式细胞术检测BC对CCA细胞周期的影响；蛋白免疫印迹（Western blot）实验分别检测BC及小干扰RNA（siRNA）敲低CCA细胞中c-Myc后对周期相关蛋白周期素依赖激酶抑制剂（p27）、细胞周期蛋白D1（Cyclin D1）及c-Myc表达的影响。结果　与0 μmol/L组相比，BC明显抑制QBC939和RBE细胞的增殖活性（P＜0.01），且高倍显微镜下观察到细胞生长减缓；BC可将QBC939细胞周期阻滞于S期（P＜0.05）；随着BC浓度的增加，p27的蛋白水平明显上调，而Cyclin D1则显著降低（P＜0.01）；BC可下调c-Myc的蛋白表达，且敲低c-Myc后p27和Cyclin D1蛋白表达的变化趋势与BC处理时一致（P＜0.05）；单独敲低c-Myc后可明显抑制QBC939和RBE细胞的存活率（P＜0.01），并能进一步增强BC的抑制作用。结论　BC通过抑制c-Myc信号通路阻滞细胞周期，进而抑制CCA细胞的增殖。     

Specific small interfering RNAs-mediated inhibition of replication of porcine encephalomyocarditis virus in BHK-21 cells

Encephalomyocarditis virus (EMCV) is recognized as a pathogen inducing acute myocarditis and sudden death in preweaned piglets and severe reproductive failure in sows. In this study, eight specific small interfering RNA (siRNA) duplexes targeting different genomic regions of EMCV BJC3 were designed and their ability to inhibit virus replication in BHK-21 cells was investigated. The results showed that BHK-21 cells transfected with siRNA duplexes to 2C gene (JH-4,666, BJC-1,739), 2B gene (BJC-807), 3C gene (BJC-2,363) and 3D gene (BJC-3269) were specifically resistant to EMCV infection when exposed to 500 times the 50% cell culture infective dose (CCID(50)) of EMCV. The levels of the 3D gene in the transfected cells were obviously decreased. IFA and Western blotting analysis confirmed that the expression of VP1 protein in cell culture transfected with the siRNAs was apparently reduced. Of the five siRNAs, JH-4,666, BJC-2,363 and BJC-3,269 were the most effective. Combination of the siRNA duplexes enhanced the inhibition of EMCV replication. Our data indicated that specific siRNAs are able to inhibit the replication of porcine encephalomyocarditis virus in BHK-21 cells, suggesting that RNAi might provide a new approach to prevent EMCV infection.     

高迁移率族蛋白A2在肿瘤发生和血管形成中的双重作用

目的:探讨高迁移率族蛋白A2(HMGA2)在肿瘤发生和血管形成中的作用。方法:采用Clontech公司软件设计靶向人HMGA2基因的5条小干扰RNA(siRNA)序列,常规培养肿瘤细胞系和血管内皮细胞系,利用脂质体2000将HM-GA2 siRNA导入细胞,通过MTT方法和Tran-swell模型检测细胞的增殖和迁移能力,采用小管形成实验检测血管内皮细胞的小管形成能力,荧光定量PCR方法检测细胞内的HMGA2 mRNA的变化。结果:5条siRNA中,HMGA2 siR-NA1、3、5不但能抑制肿瘤细胞的增殖,还可抑制血管内皮细胞的增殖、迁移和小管形成,尤其以HMGA2 siRNA5最为明显,处理组细胞的相对增长率分别为:Hela 63.2%±6.5%(P<0.01);人肺腺癌SPC-A1,86.8%±3.5%(P<0.01);人脐静脉内皮细胞(ECV-304),58.5%±6.3%(P<0.01);人膀胱上皮细胞株(HUVEC),60.3%±7.3%(P<0.01)。HMGA2 siRNA5抑制两种血管内皮细胞的迁移(P<0.01),并降低HM-GA2基因的表达(P<0.01)。结论:HMGA2除了在肿瘤发生过程中起作用之外,在血管形成中同样具有重要作用。     

Effective small interfering RNAs targeting matrix and nucleocapsid protein gene inhibit influenza A virus replication in cells and mice

RNA interference (RNAi) is a powerful tool to silence gene expression. Small interfering RNA (siRNA)-induced RNA degradation has been recently used as an antivirus agent to inhibit specific virus replication. Here, we showed that several siRNAs specific for conserved regions of influenza virus matrix (M2) and nucleocapsid protein (NP) genes could effectively inhibit expression of the corresponding viral protein. We also evaluated the antiviral potential of these siRNAs targeting M2 and NP of H5N1 avian influenza virus (AIV), which are essential to viral replication. We investigated the inhibitory effect of M2-specific siRNAs and NP-specific siRNAs on influenza A virus (H5N1, H1N1 and H9N2) replication in Madin-Darby canine kidney (MDCK) cells and BALB/c mice. The results showed that treatment with these siRNAs could specifically inhibit influenza A virus replication in MDCK cells (0.51-1.63 TCID(50) reduction in virus titers), and delivery of pS-M48 and pS-NP1383 significantly reduced lung virus titers in the infected mice (16-50-fold reduction in lung virus titers) and partially protected the mice from lethal influenza virus challenge (a survival rate of 4/8 for H1N1 virus-infected mice and 2/8 for H5N1 virus infected mice). Moreover, the treatment of pS-M48 and pS-NP1383 could suppress replication of different subtypes of influenza A viruses, including a H5N1 highly pathogenic avian isolate strain. The results provided a basis for further development of siRNA for prophylaxis and therapy of influenza virus infection in humans and animals.     

Quick identification of effective small interfering RNAs that inhibit the replication of coxsackievirus A16

Coxsackievirus A16 (CA16) is a major causative agent of hand, foot, and mouth disease (HFMD). It can cause myocarditis, pericarditis and fatal shock. There is no effective therapy against CA16. RNA interference (RNAi) is a powerful tool to silence gene expression. The small interfering RNA (siRNA) that induces RNA degradation has recently been used as an anti-virus agent to inhibit virus replication. In this study, we established the complete nucleotide sequence of CA16 strain Shzh05-1, and then compared the nucleotide sequences of Shzh05-1 with sequences of other CA16 strains in GenBank. We chose conserved regions between Shzh05-1 and the two other CA16 strains to design 30 siRNAs and construct siRNA-encoding plasmids. Thirteen siRNAs targeting conserved regions of the virus could effectively block replication of CA16 in cultured cells. Combination transfection of these 13 effective siRNAs could also produce a high inhibitory effect. These strategies and results suggest that RNAi has potential therapeutic use for suppression of CA16 infection.     

Quick identification of effective small interfering RNAs that inhibit the replication of coxsackievirus A16

Coxsackievirus A16 (CA16) is a major causative agent of hand, foot, and mouth disease (HFMD). It can cause myocarditis, pericarditis and fatal shock. There is no effective therapy against CA16. RNA interference (RNAi) is a powerful tool to silence gene expression. The small interfering RNA (siRNA) that induces RNA degradation has recently been used as an anti-virus agent to inhibit virus replication. In this study, we established the complete nucleotide sequence of CA16 strain Shzh05-1, and then compared the nucleotide sequences of Shzh05-1 with sequences of other CA16 strains in GenBank. We chose conserved regions between Shzh05-1 and the two other CA16 strains to design 30 siRNAs and construct siRNA-encoding plasmids. Thirteen siRNAs targeting conserved regions of the virus could effectively block replication of CA16 in cultured cells. Combination transfection of these 13 effective siRNAs could also produce a high inhibitory effect. These strategies and results suggest that RNAi has potential therapeutic use for suppression of CA16 infection.     

siRNA抑制人乳腺癌SK-BR-3细胞株VEGF基因表达的实验研究

目的研究siRNA(small interfering,RNA)对乳腺癌细胞SK-BR-3的VEGF基因表达的抑制作用,为RNAi技术在肿瘤生物治疗中的应用提供实验基础。方法体外合成一条针对VEGF基因的siRNA,使用脂质体转染的方法导入细胞,观察转染后乳腺癌细胞SK-BR-3的增殖变化,MTT法检测细胞存活率,RT-PCR检测转染后VEGFmRNA表达水平的变化,ELISA检测蛋白表达的下降效果。结果所设计的siRNA能有效抑制乳腺癌细胞的生长;降低了VEGFmRNA的表达;蛋白表达水平也显著降低。作为阴性对照的错义序列组siRNA则没有这种效果,不起作用。结论 siRNA可以有效抑制细胞株SK-BR-3中VEGF的表达,从而抑制细胞生长。应用RNA干扰技术可以有效抑制肿瘤细胞的增殖。     

靶向Hmga2基因的siRNA药物筛选及抗肿瘤作用

HMGA2是一种结构性转录因子,在肺癌等多种肿瘤中过表达,是肿瘤治疗的候选靶点之一。RNAi技术作为一种经济、有效的基因沉默工具,是肿瘤治疗的新手段。本文以Hmga2基因为靶点,设计并合成了5条siRNA(HMGA2 siRNA1-5),通过MTT、平板克隆、Transwell、流式细胞术等手段,研究其对肺癌细胞(NCI-H446和A549)增殖、克隆形成、迁移和凋亡等能力的影响。结果表明,HMGA2 siRNA1、3、5对肺癌细胞的各项性能具有不同程度的影响,其中HMGA2 siRNA5尤为明显。HMGA2 siRNA5主要通过沉默Hmga2基因影响细胞性能,与其干扰素效应关系不大。本文筛选获得了可有效沉默Hmga2基因的siRNA,其在体外具有良好的抗肺癌活性,是具有一定潜力的肿瘤基因治疗候选药物。