CCN3调控牙周膜成纤维细胞的增殖和凋亡  

CCN3 regulates the proliferation and apoptosis in periodontal ligament fibroblasts

作　　者：李凤霞[1] 王俊[1] 马玉云 LI Feng-xia WANG Jun MA Yu-yun.(Department of Stomatology, A nkang Hospital of Traditional Chinese Medicine. A nkang 725000, Shaanxi Province, Chin)

机构地区：[1]陕西省安康市中医医院口腔科,陕西安康725000

出　　处：《上海口腔医学》2017年第5期504-509,共6页Shanghai Journal of Stomatology

摘　　要：目的:探讨CCN3对牙周膜成纤维细胞(periodontal ligament fibroblasts,PDLFs)增殖和凋亡的作用及可能的机制。方法:构建重组载体pc DNA.3.1-CCN3并转染人PDLFs过表达CCN3,转染CCN3 si RNA,抑制其表达量。转染Fra-1 si RNA到抑制CCN3的PDLFs,同时抑制CCN3和Fra-1的表达量。应用实时荧光定量PCR(q RT-PCR)检测CCN3、Fra-1 m RNA表达水平,Western印迹法检测CCN3、Fra-1和Bcl-2蛋白表达量。MTT和Brd U实验分别检测PDLFs的生长活力和增殖能力。试剂盒方法检测caspase-3的活性。采用SPSS20.0软件包对数据进行统计学分析。结果:转染pc DNA.3.1-CCN3的实验中,CCN3 m RNA(P<0.05)和蛋白(P<0.05)表达量显著上升。转染CCN3 si RNA的实验中,CCN3 m RNA(P<0.05)和蛋白(P<0.05)表达量显著下降。CCN3表达量被抑制后,PDLFs的增殖能力(P<0.05)和生长活力(P<0.05)显著上升,caspase-3活性(P<0.05)和Bcl-2蛋白表达量(P<0.05)分别降低和升高。然而,CCN3过表达时作用相反。CCN3过表达或抑制可分别显著降低(P<0.05)或促进(P<0.01)Fra-1的表达量。另外,同时抑制CCN3和Fra-1,可促进PDLFs的凋亡(P<0.01)且抑制其增殖能力(P<0.05)。结论:抑制CCN3可通过上调Fra-1的表达,促进PDLFs的增殖能力并抑制其凋亡。PURPOSE： To investigate the effect of CCN3 on proliferation and apoptosis in periodontal ligament fibroblasts （PDLFs） and related mechanism. METHODS： Recombinant vector pcDNA.3.1-CCN3 was constructed and transfected into human PDLFs to overexpress CCN3. CCN3 siRNA was transfected to inhibit CCN3. Fra-1 siRNA was transfected into the PDLFs with CCN3 inhibition to realize the inhibition of CCN3 and Fra-1 in the meantime, mRNA expressions of CCN3 and Fra-1 were measured by quantitative real-time PCR （qRT-PCR）. The protein expressions of CCN3, Fra-land Bcl-2 were detected by Western blot. Cell growth and viability and proliferation of PDLFs were measured by 3-（4,5- Dimethyhhiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） and bromodeoxyuridine （BrdU） assays; Caspase-3 activity was tested by using the available kit. The data were analyzed with SPSS20.0 software package. RESULTS： The results showed that the mRNA （P〈0.05） and protein （P〈0.05） expressions of CCN3 were significantly up-regulated in the experimental group of pcDNA.3.I-CCN3 transfection. In addition, the mRNA （P〈0.05） and protein （P〈0.05） expressions of CCN3 were significantly decreased in the experimental group of CCN3 siRNA transfeetion. Cell growth （P〈0.05） and viability, proliferation （P〈O.05） and Bcl-2 （/9〈0.05） protein expression were increased, while caspase-3 activity （P〈0.05） decreased in the PDLFs with CCN3 inhibition. However, CCN3 overexpression exhibited reversed effect. CCN3 overexpression or inhibition could remarkably constrain （P〈0.05） or promote （P〈0.01） the expression of Fra-1, respectively. Moreover, co-inhibition of CCN3 and Fra-1 could promote apoptosis （P〈0.01） and inhibit proliferation （P〈0.05） in PDLFs. CONCLUSIONS： The results suggest that inhibition of CCN3 could accelerate proliferation and constrain apoptosis via up-regulating expression of Fra-1 in PDLFs.

关 键 词：CCN3 牙周膜成纤维细胞 Fra-1 细胞增殖 细胞凋亡 

分 类 号：R783.5[医药卫生—口腔医学]