目的研究三角形四肽重复干扰素诱导蛋白2(interferon-induced protein with tetratricopeptide repeats 2,IFIT2)在肺癌组织中的表达,并分析其表达水平与患者临床病理特征及预后的关系。方法用组织芯片技术和免疫组织化学染色法分别检...目的研究三角形四肽重复干扰素诱导蛋白2(interferon-induced protein with tetratricopeptide repeats 2,IFIT2)在肺癌组织中的表达,并分析其表达水平与患者临床病理特征及预后的关系。方法用组织芯片技术和免疫组织化学染色法分别检测90例肺腺癌、78例肺鳞癌组织,以及相应癌旁组织中IFIT2的表达,用Wilcoxon秩和检验比较肺癌及癌旁组织中IFIT2表达水平的差异,χ^2检验分析肺癌组织中IFIT2表达水平与患者临床病理特征的关系,用Kaplan-Meier生存分析法分析IFIT2表达水平与患者预后的关系,拟合Cox模型评价不同指标与患者预后的关系。结果 IFIT2在肺腺癌、肺鳞癌组织中不表达和低表达为主,在癌旁组织中高表达,在癌组织及癌旁组织中的表达差异有统计学意义(P<0.01);IFIT2染色强度与肺腺癌、肺鳞癌患者临床病理特征无相关性(P>0.05);Kaplan-Merier生存分析显示,在肺腺癌中,IFIT2低表达患者总生存期(OS)较IFIT2高表达患者短(HR=2.392,95%CI:1.103~5.186,P=0.027),多因素Cox比例风险模型显示,远处转移(HR=8.033,95%CI:3.664~17.614,P=0.000)可作为肺腺癌患者预后评判的独立风险因素;在肺鳞癌中,IFIT2低表达患者总生存期(OS)较IFIT2高表达患者短(HR=2.907,95%CI:1.118~7.559,P=0.029),多因素Cox比例风险模型显示,淋巴结转移(HR=3.390,95%CI:1.029~11.175,P=0.045)和IFIT2低表达(HR=3.762,95%CI:1.236~11.451,P=0.020)均可作为肺鳞癌患者预后评判的独立风险因素。结论 IFIT2在肺癌组织中下调表达,提示其在肺癌发生发展过程中发挥重要作用,可作为肺癌患者预后评估的重要因素。展开更多
目的探讨干扰素诱导基因IFIT1与卵巢癌免疫浸润及预后的相关性。方法通过GEO数据库筛选肿瘤免疫相关基因,Kaplan-Meier与Prognoscan预后数据库筛选与卵巢癌预后显著相关的差异基因。GEPIA、Human Protein Atlas与Timer数据库分析IFIT1...目的探讨干扰素诱导基因IFIT1与卵巢癌免疫浸润及预后的相关性。方法通过GEO数据库筛选肿瘤免疫相关基因,Kaplan-Meier与Prognoscan预后数据库筛选与卵巢癌预后显著相关的差异基因。GEPIA、Human Protein Atlas与Timer数据库分析IFIT1在卵巢癌组织与正常组织中的表达差异。UALCAN数据库分析IFIT1在不同分级和分期的卵巢癌组织中的表达情况。David数据库对String和Genemania数据库筛选的相互作用基因与蛋白进行GO富集分析。Timer和Tisidb数据库相互验证IFIT1与免疫细胞的相关性。IFIT1干扰质粒成功构建后,利用IFIT1干扰稳转株构建裸鼠移植瘤模型,观察IFIT1敲降后肿瘤生长情况,免疫组化检测中性粒细胞浸润。结果GEO数据库、Kaplan-Meier与Prognoscan预后数据库筛选出IFIT1是与卵巢癌预后显著负相关的肿瘤免疫基因。GEPIA、Human Protein Atlas、Timer数据库以及UALCAN数据库表明IFIT1在卵巢癌组织中表达显著高于正常卵巢组织(P<0.05),但与肿瘤的分期、分级并不存在显著相关性。String、Genemania、David数据库分析发现IFIT1的互作基因与蛋白富集于2’-5’寡聚腺苷酸合成酶的活化、病毒防御、先天性免疫等过程。Timer数据库表明IFIT1与卵巢癌中CD8+T细胞、B细胞、树突状细胞、中性粒细胞和巨噬细胞浸润呈正相关,其中与中性粒细胞相关性最为明显(P<0.001)。Tisidb与GSCA均验证了IFIT1与卵巢癌组织的中性粒细胞浸润呈高度正相关(P<0.05)。RT-qPCR和Western blot证明卵巢癌细胞中IFIT1被成功敲降后,IFIT1干扰细胞的移植瘤生长显著减缓(P<0.05),肿瘤中性粒细胞浸润减少。结论IFIT1可能促进中性粒细胞浸润影响卵巢癌的恶性进程。展开更多
Synthetic oligonucleotides including antisense oligonucleotides and siRNA have shown promising therapeutic potential.However,to realize the therapeutic potential of synthetic oligonucleotides,many obstacles have to be...Synthetic oligonucleotides including antisense oligonucleotides and siRNA have shown promising therapeutic potential.However,to realize the therapeutic potential of synthetic oligonucleotides,many obstacles have to be overcome,such as their poor biological stability,non-specific activity and inadequate cell membrane permeability.In this paper,the achievements by Lihe Zhang＇s group in the study of isonucleotide modified oligonucleotides and oligonucleotides conjugated with cell penetrating peptides are summarized.展开更多
RNA interference has been widely used for gene therapy of various infectious diseases and malignant tumors. However, its poor stability in serum has limited further clinic application. Here, we found that stability of...RNA interference has been widely used for gene therapy of various infectious diseases and malignant tumors. However, its poor stability in serum has limited further clinic application. Here, we found that stability of siRNA in serum enhanced obviously when 3′-terminus of sense strand （siRNA-pS） was conjugated with peptide, while same conjugation at 3′-terminus of antisense strand brought no such effects. And it was also found that only the peptide residue in siRNA-pS could be cut off by RNase A. All these results indicated that nucleases in serum prefer to invade siRNA duplex through the 3′-end of sense strand.展开更多
The therapeutic promise of small interfering RNAs （siRNAs） for specific gene silencing is dependent on the successful delivery of functional siRNAs to the cytoplasm. Their conjugation to an established delivery plat...The therapeutic promise of small interfering RNAs （siRNAs） for specific gene silencing is dependent on the successful delivery of functional siRNAs to the cytoplasm. Their conjugation to an established delivery platform, such as gold nanoparticles, offers tremendous potential for treating diseases and advancing our understanding of cellular processes. Their success or failure is dependent on both the uptake of the nanoparticles into the cells and subsequent intracellular release of the functional siRNA. In this study, utilizing gold nanoparticle siRNA-mediated delivery against C-MYC, we aimed to determine if we could achieve knockdown in a cancer cell line with low levels of intracellular glutathione, and determine the influence, if any, of polyethylene glycol （PEG） ligand density on knockdown, with a view to determining the optimal nanoparticle design to achieve C-MYC knockdown. We demonstrate that, regardless of the PEG density, knockdown in cells with relatively low glutathione levels can be achieved, as well as the possible effect of steric hindrance of PEG on the availability of the siRNA for cleavage in the intracellular environment. Gold nanoparticle uptake was demonstrated via transmission electron microscopy and mass spectroscopy, while knockdown was determined at the protein and physiological levels （cells in S-phase） by in-cell westerns and BrdU incorporation, respectively.展开更多
文摘目的研究三角形四肽重复干扰素诱导蛋白2(interferon-induced protein with tetratricopeptide repeats 2,IFIT2)在肺癌组织中的表达,并分析其表达水平与患者临床病理特征及预后的关系。方法用组织芯片技术和免疫组织化学染色法分别检测90例肺腺癌、78例肺鳞癌组织,以及相应癌旁组织中IFIT2的表达,用Wilcoxon秩和检验比较肺癌及癌旁组织中IFIT2表达水平的差异,χ^2检验分析肺癌组织中IFIT2表达水平与患者临床病理特征的关系,用Kaplan-Meier生存分析法分析IFIT2表达水平与患者预后的关系,拟合Cox模型评价不同指标与患者预后的关系。结果 IFIT2在肺腺癌、肺鳞癌组织中不表达和低表达为主,在癌旁组织中高表达,在癌组织及癌旁组织中的表达差异有统计学意义(P<0.01);IFIT2染色强度与肺腺癌、肺鳞癌患者临床病理特征无相关性(P>0.05);Kaplan-Merier生存分析显示,在肺腺癌中,IFIT2低表达患者总生存期(OS)较IFIT2高表达患者短(HR=2.392,95%CI:1.103~5.186,P=0.027),多因素Cox比例风险模型显示,远处转移(HR=8.033,95%CI:3.664~17.614,P=0.000)可作为肺腺癌患者预后评判的独立风险因素;在肺鳞癌中,IFIT2低表达患者总生存期(OS)较IFIT2高表达患者短(HR=2.907,95%CI:1.118~7.559,P=0.029),多因素Cox比例风险模型显示,淋巴结转移(HR=3.390,95%CI:1.029~11.175,P=0.045)和IFIT2低表达(HR=3.762,95%CI:1.236~11.451,P=0.020)均可作为肺鳞癌患者预后评判的独立风险因素。结论 IFIT2在肺癌组织中下调表达,提示其在肺癌发生发展过程中发挥重要作用,可作为肺癌患者预后评估的重要因素。
文摘目的探讨干扰素诱导基因IFIT1与卵巢癌免疫浸润及预后的相关性。方法通过GEO数据库筛选肿瘤免疫相关基因,Kaplan-Meier与Prognoscan预后数据库筛选与卵巢癌预后显著相关的差异基因。GEPIA、Human Protein Atlas与Timer数据库分析IFIT1在卵巢癌组织与正常组织中的表达差异。UALCAN数据库分析IFIT1在不同分级和分期的卵巢癌组织中的表达情况。David数据库对String和Genemania数据库筛选的相互作用基因与蛋白进行GO富集分析。Timer和Tisidb数据库相互验证IFIT1与免疫细胞的相关性。IFIT1干扰质粒成功构建后,利用IFIT1干扰稳转株构建裸鼠移植瘤模型,观察IFIT1敲降后肿瘤生长情况,免疫组化检测中性粒细胞浸润。结果GEO数据库、Kaplan-Meier与Prognoscan预后数据库筛选出IFIT1是与卵巢癌预后显著负相关的肿瘤免疫基因。GEPIA、Human Protein Atlas、Timer数据库以及UALCAN数据库表明IFIT1在卵巢癌组织中表达显著高于正常卵巢组织(P<0.05),但与肿瘤的分期、分级并不存在显著相关性。String、Genemania、David数据库分析发现IFIT1的互作基因与蛋白富集于2’-5’寡聚腺苷酸合成酶的活化、病毒防御、先天性免疫等过程。Timer数据库表明IFIT1与卵巢癌中CD8+T细胞、B细胞、树突状细胞、中性粒细胞和巨噬细胞浸润呈正相关,其中与中性粒细胞相关性最为明显(P<0.001)。Tisidb与GSCA均验证了IFIT1与卵巢癌组织的中性粒细胞浸润呈高度正相关(P<0.05)。RT-qPCR和Western blot证明卵巢癌细胞中IFIT1被成功敲降后,IFIT1干扰细胞的移植瘤生长显著减缓(P<0.05),肿瘤中性粒细胞浸润减少。结论IFIT1可能促进中性粒细胞浸润影响卵巢癌的恶性进程。
基金National Natural Science Foundation (Grant No.20932001)the Ministry of Science and Technology (Grant No.2012CB720604)
文摘Synthetic oligonucleotides including antisense oligonucleotides and siRNA have shown promising therapeutic potential.However,to realize the therapeutic potential of synthetic oligonucleotides,many obstacles have to be overcome,such as their poor biological stability,non-specific activity and inadequate cell membrane permeability.In this paper,the achievements by Lihe Zhang＇s group in the study of isonucleotide modified oligonucleotides and oligonucleotides conjugated with cell penetrating peptides are summarized.
基金Ministry of Science and Technology of China(Grant No.2012CB720604)National Natural Science Foundation of China(Grant No.20932001)
文摘RNA interference has been widely used for gene therapy of various infectious diseases and malignant tumors. However, its poor stability in serum has limited further clinic application. Here, we found that stability of siRNA in serum enhanced obviously when 3′-terminus of sense strand （siRNA-pS） was conjugated with peptide, while same conjugation at 3′-terminus of antisense strand brought no such effects. And it was also found that only the peptide residue in siRNA-pS could be cut off by RNase A. All these results indicated that nucleases in serum prefer to invade siRNA duplex through the 3′-end of sense strand.
文摘The therapeutic promise of small interfering RNAs （siRNAs） for specific gene silencing is dependent on the successful delivery of functional siRNAs to the cytoplasm. Their conjugation to an established delivery platform, such as gold nanoparticles, offers tremendous potential for treating diseases and advancing our understanding of cellular processes. Their success or failure is dependent on both the uptake of the nanoparticles into the cells and subsequent intracellular release of the functional siRNA. In this study, utilizing gold nanoparticle siRNA-mediated delivery against C-MYC, we aimed to determine if we could achieve knockdown in a cancer cell line with low levels of intracellular glutathione, and determine the influence, if any, of polyethylene glycol （PEG） ligand density on knockdown, with a view to determining the optimal nanoparticle design to achieve C-MYC knockdown. We demonstrate that, regardless of the PEG density, knockdown in cells with relatively low glutathione levels can be achieved, as well as the possible effect of steric hindrance of PEG on the availability of the siRNA for cleavage in the intracellular environment. Gold nanoparticle uptake was demonstrated via transmission electron microscopy and mass spectroscopy, while knockdown was determined at the protein and physiological levels （cells in S-phase） by in-cell westerns and BrdU incorporation, respectively.
