ADAM28反义核酸对人牙龈成纤维细胞生物学功能的影响

The effects of ADAM28 AS-ODN on biological function of human gingival fibroblasts

目的:研究金属蛋白酶解离素28(a disintegrin and metalloproteinase 28,ADAM28)反义核酸(antisense oligodeoxynucleotide,AS-ODN)对人牙龈成纤维细胞(human gingival fibroblasts,HGFs)增殖、分化、凋亡的影响,并分析可能的作用机制。方法:采用细胞培养、基因转染、四甲基唑蓝(MTT)比色法、酶动力学法和流式细胞术(FCM)检测ADAM28反义核酸转染HGFs后对细胞生物学特性的影响。采用SPSS16.0软件包中的SNK检验对数据进行统计学分析。结果:ADAM28 AS-ODN组HGFs的增殖活性显著降低。AS-ODN组HGFs处于S期的细胞百分比显著低于SODN组和未转染组,G2+M期的细胞百分比显著低于未转染组,AS-ODN组的细胞增殖指数(PI=S+G2M)显著低于其他2组,差异显著。AS-ODN组碱性磷酸酶(ALP)分泌活性、凋亡细胞百分比显著升高。结论:ADAM28反义核酸可显著抑制HGFs的增殖,并影响细胞周期的变化,促进其分化,显著诱导HGFs的凋亡。

PURPOSE： To study the effects of a disintegrin and metalloproteinase 28 （ADAM28） antisense oligodeoxynueleotide （AS-ODN） on proliferation, differentiation and apoptosis of human gingival fibroblasts （HGFs）, and analyze the possible mechanism. METHODS： Cell culture, gene transfection, MTT chromatometry, enzyme dynamics, and flow cytometry （FCM） techniques were used to detect the effects of ADAM28 AS-ODN on biological characteristics of HGFs after transfected into HGFs. The statistical differences were evaluated by SNK test with SPSS 16.0 software package. RESULTS： In ADAM28 AS-ODN group, the proliferation activity of HGFs decreased significantly. Cell percentage in S phase in AS-ODN group was notably lower than that of S-ODN and untransfected groups, and cell percentage in G2＋M phase was remarkably lower than that of untransfeeted group. Cell proliferation index （PI=S＋G2M）in AS-ODN group was significantly lower than that of the other two groups. There was a significant difference between the groups. In AS-ODN group, alkaline phosphatase （ALP） secretion activity and percentage of apoptotic cell notably increased. CONCLUSIONS： ADAM28 AS-ODN could inhibit HGFs proliferation significantly and influence the changes of cell cycle, promote HGFs differentiation and induce HGFs apoptosis remarkably.Supported by Chinese Post-Doctoral Special Science Foundation （201003774）.