N-M Ng1, S-P Jiang, Z-Q Lv. 1. Clinical Department of School of Medicine, Shenzhen University, Shenzhen, China.
Abstract
BACKGROUND AND OBJECTIVE: Calcium signaling is important for both normal physiologic processes and pathology of various diseases. Transient receptor potential melastatin 7 (TRPM7) gene has been reported to be a potential candidate for calcium influx. The present study aimed to investigate the possible role of TRPM7 channels in apoptosis in rat basophilic leukemia mast cell line (RBL-2H3), which is widely used in mast cell-associated studies. MATERIALS AND METHODS: A recombinant retrovirus vector siRNA targeting rat TRPM7 gene was constructed and identified. Cellular survival was assessed by MTT. Cell apoptosis was evaluated by flow cytometry and TUNEL-FITC/Hoechst 33258 staining. RESULTS: The transfection efficiency by retrovirus vector was about 60%-70%. Transfection with TRPM7 siRNA significantly reduced TRPM7 expression both at mRNA and protein levels. Suppression of TRPM7 expression by siRNA led to significantly decreased cellular survival rates and increased apoptosis rates in RBL-2H3 cells. CONCLUSIONS: This study indicates that TRPM7 is involved in the apoptosis process in RBL-2H3 cells.
BACKGROUND AND OBJECTIVE:Calcium signaling is important for both normal physiologic processes and pathology of various diseases. Transient receptor potential melastatin 7 (TRPM7) gene has been reported to be a potential candidate for calcium influx. The present study aimed to investigate the possible role of TRPM7 channels in apoptosis in rat basophilic leukemia mast cell line (RBL-2H3), which is widely used in mast cell-associated studies. MATERIALS AND METHODS: A recombinant retrovirus vector siRNA targeting ratTRPM7 gene was constructed and identified. Cellular survival was assessed by MTT. Cell apoptosis was evaluated by flow cytometry and TUNEL-FITC/Hoechst 33258 staining. RESULTS: The transfection efficiency by retrovirus vector was about 60%-70%. Transfection with TRPM7 siRNA significantly reduced TRPM7 expression both at mRNA and protein levels. Suppression of TRPM7 expression by siRNA led to significantly decreased cellular survival rates and increased apoptosis rates in RBL-2H3 cells. CONCLUSIONS: This study indicates that TRPM7 is involved in the apoptosis process in RBL-2H3 cells.
Authors: Saied Froghi; Charlotte R Grant; Radhika Tandon; Alberto Quaglia; Brian Davidson; Barry Fuller Journal: Clin Rev Allergy Immunol Date: 2021-01-06 Impact factor: 8.667