Purpose:To investigate the effect of small interfering RNA (siRNA)targeting VEGF of retinal pigment epithelium (RPE) cells on the growth activity of human retinal vascular endothelial cells (RECs) under a co-culture s...Purpose:To investigate the effect of small interfering RNA (siRNA)targeting VEGF of retinal pigment epithelium (RPE) cells on the growth activity of human retinal vascular endothelial cells (RECs) under a co-culture system.Methods:By applying the vector.(pGPU6)-based siRNA plasmid gene silence system,we specifically silenced VEGF expression of RPE cells (ARPE-19) through plasmid (pG-PU6-VEGFA-siRNA)transfection.Reverse transcription polymerase chain reaction(RT-PCR) was applied for selecting the most efficient siRNA segment among three pGPU6-VEGF-siRNA groups (siRNA-1,siRNA-2 and siRNA-3).Treated RPE cells were co-cultured with RECs in a co-culture system made up of a 24-well culture plate and transwell inserts assembled inside During 7-day culture period,the growth ca-pacity of RECs were observed and tested in the form of cell counting assay.Three groups were established in this study:RPE cells transfected with pGPU6-VEGF-siRNA and co-cultured with RECs (group A),RPE cells transfected with siR-NA null vector and co-cultured with RECs (group B),and RECs cultured alone (group C).Results:After transfection,VEGF expression levels of RPE cells in three pGPU6-VEGF-siRNA groups (siRNA-1,siR-NA-2 and siRNA-3)evaluated by RT-PCR were 2.56 ±0.45,1.17 ±0.38 and 4.39 ±0.51,respectively (n =10).siRNA-2 was selected as the foremost segment for transfection (P < 0.05,SNK-q test).During the 7-day co-culture period,the influence upon the growth of RECs was observed.Growth curve of RECs under co-culture showed a lower growth rate in group A than in group B (P <0.05,dunnett's test),but no significant difference between group A and group C was noted (P>0.05,dunnett's test).RECs in group A proliferated muchfaster during the first four days post-transfection.Conclusion:Delivery of siRNA targeting VEGF plays an efficient role in down-regulating VEGF expression in RPE cells,therefore modulating the growth activity of RECs under a co-culture system in vitro.The application of this technique may provide novel evidence for the prevention and treatment of retinal neovascularization diseases.展开更多
目的:探讨核因子-κB(NF-κB)和血管内皮生长因子(VEGF)在胃癌中的表达及其临床意义.方法:选取68例胃癌组织为实验对象,各对应的癌旁组织为对照组,并选取20例正常胃组织为正常对照组.采用免疫组织化学法(SABC)检测各样本的NF-κB和VEGF...目的:探讨核因子-κB(NF-κB)和血管内皮生长因子(VEGF)在胃癌中的表达及其临床意义.方法:选取68例胃癌组织为实验对象,各对应的癌旁组织为对照组,并选取20例正常胃组织为正常对照组.采用免疫组织化学法(SABC)检测各样本的NF-κB和VEGF,并对相关的临床病理资料进行统计分析.结果:正常胃组织中未见NF-κB阳性表达及VEGF表达,胃癌细胞中阳性表达率均明显高于癌旁组织中(63.2% vs 2.9%,76.5% vs 14.7%,均P=0.000).NF-κB及VEGF在胃癌组织中的表达均与性别、年龄、组织学类型无关;NF-κB及VEGF在TNM分期(Ⅲ+Ⅳ期)、有淋巴结转移、累及浆膜、有远处转移组阳性表达均明显高于TNM(Ⅰ+Ⅱ期)、无淋巴结转移、未累及浆膜、无远处转移组(P<0.05).胃癌中NF-κB与VEGF表达显著正相关(r=0.512,P=0.000).结论:NF-κB和VEGF在胃癌中呈现高表达,且两者存在明显相关,NF-κB可能通过上调VEGF的表达促进胃癌的发生发展.展开更多
目的观察中药川贝含药血清对支气管平滑肌细胞(BSMC)增殖,管道形成及血管内皮细胞生长因子(VEGF)表达的影响。方法卵蛋白(OVA)复制哮喘小鼠模型,制备川贝含药血清。体外培养BSMC,随机分为空白血清组,低剂量组和高剂量组。检测BSMC增殖率...目的观察中药川贝含药血清对支气管平滑肌细胞(BSMC)增殖,管道形成及血管内皮细胞生长因子(VEGF)表达的影响。方法卵蛋白(OVA)复制哮喘小鼠模型,制备川贝含药血清。体外培养BSMC,随机分为空白血清组,低剂量组和高剂量组。检测BSMC增殖率,管道形成长度,VEGF及VEGF m RNA的表达。结果与空白血清组比较,低剂量组和高剂量组显著降低BSMC增殖,管道形成长度,VEGF和VEGF m RNA的表达(P<0.05)。结论中药川贝含药血清可抑制BSMC增殖,管道形成长度,其机制可能与其抑制VEGF的表达有关。展开更多
文摘Purpose:To investigate the effect of small interfering RNA (siRNA)targeting VEGF of retinal pigment epithelium (RPE) cells on the growth activity of human retinal vascular endothelial cells (RECs) under a co-culture system.Methods:By applying the vector.(pGPU6)-based siRNA plasmid gene silence system,we specifically silenced VEGF expression of RPE cells (ARPE-19) through plasmid (pG-PU6-VEGFA-siRNA)transfection.Reverse transcription polymerase chain reaction(RT-PCR) was applied for selecting the most efficient siRNA segment among three pGPU6-VEGF-siRNA groups (siRNA-1,siRNA-2 and siRNA-3).Treated RPE cells were co-cultured with RECs in a co-culture system made up of a 24-well culture plate and transwell inserts assembled inside During 7-day culture period,the growth ca-pacity of RECs were observed and tested in the form of cell counting assay.Three groups were established in this study:RPE cells transfected with pGPU6-VEGF-siRNA and co-cultured with RECs (group A),RPE cells transfected with siR-NA null vector and co-cultured with RECs (group B),and RECs cultured alone (group C).Results:After transfection,VEGF expression levels of RPE cells in three pGPU6-VEGF-siRNA groups (siRNA-1,siR-NA-2 and siRNA-3)evaluated by RT-PCR were 2.56 ±0.45,1.17 ±0.38 and 4.39 ±0.51,respectively (n =10).siRNA-2 was selected as the foremost segment for transfection (P < 0.05,SNK-q test).During the 7-day co-culture period,the influence upon the growth of RECs was observed.Growth curve of RECs under co-culture showed a lower growth rate in group A than in group B (P <0.05,dunnett's test),but no significant difference between group A and group C was noted (P>0.05,dunnett's test).RECs in group A proliferated muchfaster during the first four days post-transfection.Conclusion:Delivery of siRNA targeting VEGF plays an efficient role in down-regulating VEGF expression in RPE cells,therefore modulating the growth activity of RECs under a co-culture system in vitro.The application of this technique may provide novel evidence for the prevention and treatment of retinal neovascularization diseases.
文摘目的:探讨核因子-κB(NF-κB)和血管内皮生长因子(VEGF)在胃癌中的表达及其临床意义.方法:选取68例胃癌组织为实验对象,各对应的癌旁组织为对照组,并选取20例正常胃组织为正常对照组.采用免疫组织化学法(SABC)检测各样本的NF-κB和VEGF,并对相关的临床病理资料进行统计分析.结果:正常胃组织中未见NF-κB阳性表达及VEGF表达,胃癌细胞中阳性表达率均明显高于癌旁组织中(63.2% vs 2.9%,76.5% vs 14.7%,均P=0.000).NF-κB及VEGF在胃癌组织中的表达均与性别、年龄、组织学类型无关;NF-κB及VEGF在TNM分期(Ⅲ+Ⅳ期)、有淋巴结转移、累及浆膜、有远处转移组阳性表达均明显高于TNM(Ⅰ+Ⅱ期)、无淋巴结转移、未累及浆膜、无远处转移组(P<0.05).胃癌中NF-κB与VEGF表达显著正相关(r=0.512,P=0.000).结论:NF-κB和VEGF在胃癌中呈现高表达,且两者存在明显相关,NF-κB可能通过上调VEGF的表达促进胃癌的发生发展.
文摘目的观察中药川贝含药血清对支气管平滑肌细胞(BSMC)增殖,管道形成及血管内皮细胞生长因子(VEGF)表达的影响。方法卵蛋白(OVA)复制哮喘小鼠模型,制备川贝含药血清。体外培养BSMC,随机分为空白血清组,低剂量组和高剂量组。检测BSMC增殖率,管道形成长度,VEGF及VEGF m RNA的表达。结果与空白血清组比较,低剂量组和高剂量组显著降低BSMC增殖,管道形成长度,VEGF和VEGF m RNA的表达(P<0.05)。结论中药川贝含药血清可抑制BSMC增殖,管道形成长度,其机制可能与其抑制VEGF的表达有关。
