Role of ANX A2 in APL-induced expression of tissue factor in THP-1 cells
- VernacularTitle:ANX A2在抗磷脂抗体诱导THP-1细胞组织因子表达中的作用
- Author:
Na LI
;
Hong ZHOU
;
Ying YU
;
Ting WANG
;
Hongliang HUANG
;
Wenxia SHI
;
Haibo WANG
- Publication Type:Journal Article
- Keywords:
RNA interference;
Lentiviral vector;
Annexin A2;
Antiphospholipid antibodies;
Tissue factor
- From:
Chinese Journal of Pathophysiology
2009;25(12):2447-2453
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To construct a lentiviral vector harboring RNAi sequence targeting human annexin A2 (ANX A2) and to investigate the functions of ANX A2 in antiphospholipid antibody (APL)-induced tissue factor (TF) expression in monocytes. METHODS: Four different short hairpin RNAs (shRNA) targeting ANX A2 gene were constructed and cloned into the pGCSIL-GFP vector. After identification with PCR and sequencing, the effective interference sequence was further determined by Western blotting analysis. The recombinant lentivirus LV-RNAi-ANX A2 harvested from 293T cells was transferred into THP-1 cells and the ANX A2 mRNA and protein expression on THP-1 cells were examined. The transferred-THP-1 cells were stimulated by APL/β_2GPI compound, and the TF mRNA and TF activity was assayed. RESULTS: The RNAi sequences targeting the human ANX A2 gene were successfully inserted into the lentiviral vector and the high-performance RNA interference sequence was sieved out. The recombinant lentivirus was harvested from 293T cells with a viral titer of 3×10~(12) TU/L. THP-1 cells infected with LV-RNAi-ANX A2 showed almost lockout of ANX A2 both at mRNA and protein levels. The TF expression was also significantly decreased in the transferred-THP-1 cells induced by APL/β_2GPI compound. CONCLUSION: The lentiviral vector constructed in the present study blocks the ANX A2 expression in THP-1 cells and further inhibits the TF expression induced by APL/β_2GPI compound, which indicates that ANX A2 do play an important role in APL-induced TF expression on monocytes.
