【摘要】 目的探讨长链非编码RNA(LncRNAs)核仁小分子RNA宿主基因11(SNHG11)对高糖诱导的人视网膜色素上皮细胞(ARPE-19)上皮间质转化(EMT)及增殖、迁移的影响。方法将ARPE-19细胞分为低糖对照组(5.5 mmol/L葡萄糖处理48 h)、高渗对照组(60.0 mmol/L甘露醇处理48 h)以及高糖干预组(60.0 mmol/L葡萄糖处理48 h)、si-NC组(将si-conSNHG11转染至ARPE-19细胞以后用60.0 mmol/L葡萄糖处理48 h)、siSNHG11组(将siSNHG11转染至ARPE-19细胞后用60.0 mmol/L葡萄糖处理48 h)。采用实时荧光定量PCR检测各组细胞LncRNA SNHG11的表达量;采用Western blot方法检测各组细胞中EMT标志物E-钙黏蛋白(E-cad)、紧密连接蛋白(ZO-1)、波形蛋白(Vimentin)、α-平滑肌肌动蛋白(α-SMA)的表达;采用细胞划痕法检测各组细胞迁移面积;采用CCK-8法检测各组细胞存活率。结果实时荧光定量PCR检测结果显示,SNHG11在高糖干预组细胞中表达量显著高于低糖对照组及高渗对照组(F=44.27,P<0.05)。高糖干预组细胞中E-cad及ZO-1表达量均低于低糖对照组及高渗对照组(F=124.41、5.66,P<0.05);Vimentin及α-SMA表达量均高于低糖对照组及高渗对照组(F=8.37、5.48,P<0.05)。Si-SNHG11组细胞中E-cad及ZO-1表达量均高于si-NC组(t=3.15、3.14),P<0.05);Vimentin及α-SMA表达量均低于si-NC组(t=3.30、13.44),P<0.05)。细胞划痕实验显示,高糖干预组48 h迁移面积显著大于低糖对照组及高渗对照组(F=9.142,P<0.05);si-SNHG11组48 h迁移面积显著小于Si-NC组(t=10.094,P<0.05)。CCK-8法检测显示,高糖干预组细胞存活率显著高于低糖对照组及高渗对照组(F=7.713,P<0.05);Si-SNHG11组细胞的存活率显著低于Si-NC组(t=5.371,P<0.05)。结论 LncRNA SNHG11在高糖诱导的人视网膜色素上皮细胞中表达上调;沉默SNHG11表达可抑制人视网膜色素上皮细胞的EMT、增殖及迁移。更多还原

【Abstract】 Objective To investigate the effect of the long non-coding RNA(LncRNA) small nucleolar RNA host gene 11(SNHG11) on the epithelial-mesenchymal transition(EMT), proliferation, and migration of human retinal pigment epithelial cells(ARPE-19) induced by high glucose. Methods ARPE-19 cells were divided into low-glucose control group(treated with 5.5 mmol/L glucose for 48 h), hypertonic control group(treated with 60.0 mmol/L mannitol for 48 h), high-glucose intervention group(treated with 60.0 mmol/L glucose for 48 h), si-NC group(ARPE-19 cells were transfected with si-conSNHG11 and then treated with 60.0 mmol/L glucose for 48 h), and si-SNHG11 group(ARPE-19 cells were transfected with siSNHG11 and then treated with 60.0 mmol/L glucose for 48 h). Quantitative real-time PCR was used to measure the expression of LncRNA SNHG11 in each group; Western blot was used to measure the expression of the EMT markers E-cadherin(E-cad), zonula occluden-1(ZO-1), vimentin, and α-smooth muscle actin(α-SMA); wound healing assay was used to measure cell migration area; CCK-8 assay was used to measure the survival rate of cells. Results Quantitative real-time PCR showed that the high-glucose intervention group had a significantly higher expression level of LncRNA SNHG11 than the low-glucose control group and the hypertonic control group(F=44.27,P<0.05). Compared with the low-glucose control group and the hypertonic control group, the high-glucose intervention group had significantly lower expression levels of E-cad and ZO-1(F=124.41,5.66,P<0.05) and a significantly hig-her expression level of vimentin and α-SMA(F=8.37,5.48,P<0.05). Compared with the si-NC group, the si-SNHG11 group had significantly higher expression levels of E-cad and ZO-1(t=3.15,3.14,P<0.05) and significantly lower expression levels of vimentin and α-SMA(t=3.30,13.44,P<0.05). Wound healing assay showed that after 48 h of treatment, the high-glucose intervention group had a significantly larger migration area than the low-glucose control group and the hypertonic control group(F=9.142,P<0.05), and the si-SNHG11 group had a significantly smaller migration area than the si-NC group(t=10.094,P<0.05). CCK-8 assay showed that the high-glucose intervention group had a significantly higher survival rate of cells than the low-glucose control group and the hypertonic control group(F=7.713,P<0.05), and the si-SNHG11 group had a significantly lower survival rate of cells than the si-NC group(t=5.371,P<0.05). Conclusion The expression of LncRNA SNHG11 is upregulated in human re-tinal pigment epithelial cells induced by high glucose, and silencing the expression of SNHG11 can inhibit the EMT, proliferation, and migration of human retinal pigment epithelial cells.更多还原