ADAM28反义核酸对人牙龈成纤维细胞生物学功能的影响

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S-ODN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。

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

PURPOSE: To study the effects of a disintegrin and metalloproteinase 28 (ADAM28) antisense oligodeoxynucleotide (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 untransfected 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.