钙结合蛋白S100A11对人静脉畸形组织和人脐静脉内皮细胞增殖、血管生成和局部侵袭能力的影响

Effect of calcium binding protein S100A11 on proliferation, angiogenesis and local invasion of human venous malformation and human umbilical vein endothelial cells

目的观察对钙结合蛋白S100A11在人静脉畸形组织和人脐静脉内皮细胞(HUVEC)中生物学特性的研究,探讨其对静脉畸形组织和HUVEC的增殖、血管生成、局部侵袭等生物学能力的影响。方法将静脉组织研磨后提取总RNA,实时荧光定量聚合酶链式反应(RT-qPCR)检测人静脉畸形组织和对照正常血管组织标本中S100A11的相对表达量。将HUVEC细胞随机分为实验组和对照组,即si-S100A11组和si-NC组,利用RNA干扰技术沉默si-S100A11组编码S100A11蛋白mRNA。通过RT-qPCR分别检测两组细胞的转染效率;细胞计数试剂盒(CCK-8)检测两组细胞的增殖活性;Transwell检测两组细胞的局部侵袭能力;Matrigel血管生成实验检测沉默后细胞的成管数目。两组间比较采用两样本独立t检验。结果RT-qPCR结果表明,静脉畸形组织(VMs组)与正常血管组织(NC组)中S100A11的表达量分别为14.97±16.30和1.01±0.01,差异有统计学意义(t=-3.63,P<0.01);RNA干扰后的si-S100A11组和si-NC组S100A11的mRNA相对表达量分别为0.25±0.02和1.97±0.03,差异有统计学意义(t=73.33,P<0.01);CCK-8实验结果表明,HUVECs在处理后48 h si-S100A11组与si-NC组的相对吸光度值分别为0.74±0.00和0.86±0.01,差异有统计学意义(t=19.82,P<0.05);处理后72 h si-S100A11组和si-NC组的相对吸光度值分别为0.85±0.00和1.08±0.01,差异有统计学意义(t=16.50,P<0.01);Transwell局部侵袭的细胞数量si-S100A11组和si-NC组分别为(114.67±19.55)个和(566.67±113.76)个,差异有统计学意义(t=6.78,P<0.01);血管生成实验si-S100A11组和si-NC组总成管数目分别为(56.67±3.48)个和(149.00±5.77)个,差异有统计学意义(t=13.70,P<0.01)。结论沉默S100A11后HUVECs的增殖、血管生成、局部侵袭能力受到抑制,S100A11可能对静脉畸形增殖、血管生成、局部侵袭等生物学行为均有影响。

Objective To investigate the biological characteristics of S100A1l,a calcium-binding human venous malformation and human umbilical vein endothelial cells(HUVEC),and to elucidate its effects on HUVEC proliferation,migration,invasion,and angiogenesis.Methods Pathological specimens were collected from 18 patients with surgically removed venous malformations at the Third Affiliated Hospital of Zhengzhou University.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)was employed to determine the relative expression of S100A11 in human venous malformation tissues compared to normal control vascular tissues.The cells were divided into an experimental group(si-S100A11 group)and a control group(si-NC group),where RNA interference technology was used to silence the mRNA encoding S10OA11 protein.The transfection efficiency of both groups was assessed by RT-qPCR.Cell proliferative activity was evaluated using the cell counting kit-8(CCK-8)cell counting kit.Migration and invasion abilities were examined through Transwell assays.Matrigel angiogenesis assay was conducted to measure tub formation after silencing.Results The results of RT-qPCR showed that the expression level of Si00A1l in venous malformation tissues(VMs group)and normal vascular tissues(NC group)was 14.97±16.30 and 1.01±0.01,respectively,and the difference was statistically significant(t=-3.63.P<0.01);The mRNA relative expression level of S100A11 in si-S100A11 group and si-NC group after RNA interference was 0.25±0.02 and 1.97±0.03,respectively,and the difference was statistically significant(t=73.33,P<0.01);The results of CCK-8 experiment showed that the relative light absorption values of HUVECs in the si-S100A11 group and the si-NC group were 0.74±0.00 and 0.86±0.01,respectively,48 h after treatment,and the difference was statistically significant(t=19.82,P<0.05);After 72 h treatment,the relative light absorption values of si-S100A1l group and si-NC group were 0.85±0.00 and 1.08±0.01,respectively,and the difference was statistically significant(t=16.50,P<0.01);The number of Transwell locally invaded cells in si-S100A11 group and si-NC group was(114.67±19.55)and(566.67±113.76),respectively,and the difference was statistically significant(t=6.78,P<0.01):The number of assembly tubes in the si-S100A11 group and the si-NC group was(56.67±3.48)and(149.00±5.77),respectively,and the difference was statistically significant(t=13.70,P<0.01).Conclusion After silencing S10OA1l,the proliferation,angiogenesis and local invasion of HUVECs are inhibited,and S100A1l may have effects on the biological behavior of venous malformation,proliferation,angiogenesis and local invasion.