Effect of CaMKIIγ RNA interference on the expression of NFATc1,TRAP and c-Src genes in osteoclast differentiation
10. 11904/j. issn. 1002-3070. 2019. 04. 002
- VernacularTitle:CaMKⅡγ基因沉默对破骨细胞分化中NFATc1 、TRAP、c-Src基因表达的影响
- Author:
Mengnan LIU
1
;
Hui WANG
;
Mengchun QI
;
Wei DONG
;
Ren LI
;
Hong SUN
Author Information
1. 华北理工大学口腔医学院口腔颌面外科教研室(唐山 063000)
- Keywords:
Calmodulin-dependent kinase IIγ;
RNA interference;
Nuclear factor of activated T-cells cytoplasmic 1;
Cell-sarcoma receptor coactivator;
Tartrate resistant acid phosphatase
- From:Practical Oncology Journal
2019;33(4):294-299
- CountryChina
- Language:Chinese
-
Abstract:
Objective The aim of this study was to investigate the effect of Ca2+ /calmodulin - dependent kinase II (CaMKII)γ RNA interference on the expression of nuclear factor of activated T-cells cytoplasmic 1(NFATc1),tyrosine kinase(c-Src)and tartrate resistant acid phosphatase(TRAP)genes,and its role and molecular mechanism in osteoclast differentiation. Methods The CaMKII γ RNA interference vector was constructed by lentivirus and transfected into RAW264. 7 cells. The experiment was di-vided into three groups:A,B and C,which were the control group,negative vector group and interference vector group. After transfec-tion for 12 hours,osteoclasts induced by 50 ng/mL RANKL and the cells were harvested after induction for 5 days. Real-time quanti-tative PCR,Western blot and immunofluorescence were used to detect the expression of NFATc1,TRAP and c-Src genes in three groups. Results The mRNA levels of NFATc1,TRAP and c-Src in the group C decreased by 49. 86% ,43. 65% and 53. 57% ,re-spectively(P<0. 001),and the protein levels decreased by 54. 22% ,46. 75% and 45. 86% ,respectively(P<0. 001). There was no significant difference between the A and the B groups(P>0. 05). The fluorescence intensity of the above genes in the group C was significantly weaker than that in the A and B groups,and the formation of osteoclasts was significantly less than that in the A and B groups. Conclusion CaMKIIγ RNA interference significantly inhibited the expression of NFATc1,TRAP and c-Src genes,sugges-ting that CaMKIIγ plays a key regulatory role in osteoclast differentiation.
