瞬时感受器电位M7通道对RBL-2H3细胞增殖及凋亡的影响

Role of Transient Receptor Potential Melastatin 7 in Proliferation and Apoptosis of RBL-2H3 Cells

【目的】观察瞬时感受器电位M7通道(TRPM7)在大鼠肥大细胞系RBL-2H3的表达,探讨其对RBL-2H3增殖和凋亡的影响。【方法】Western blot及细胞免疫荧光分别检测TRPM7在RBL-2H3的表达。通过离子通道阻断剂2-APB抑制TRPM7通道的表达,以WST-1法检测细胞增殖,流式细胞术及Hoechst33342荧光染色检测细胞凋亡。【结果】Westernblot及细胞免疫荧光法均检测到TRPM7在RBL-2H3的表达,细胞免疫荧光提示TRPM7主要表达于RBL-2H3细胞膜。100μmol/L和200μmol/L的2-APB可明显抑制TRPM7的表达。100μmol/L和200μmol/L的2-APB对RBL-2H3的增殖抑制率分别为(30.4±4.2)%和(42.0±0.8)%,早期凋亡率分别为(36.9±6.7)%和(49.3±1.8)%,总凋亡率分别为(40.2±7.0)%和(76.8±5.5)%。【结论】TRPM7通道表达于RBL-2H3肥大细胞系并参与其增殖和凋亡。

【Objective】To investigate the expression of transient receptor potential melastatin 7（TRPM7）and its role in proliferation and apoptosis in rat basophilic leukemia cell line（RBL-2H3）.【Methods】Expression of TRPM7 in RBL-2H3 were detected by Western blot and immunofluorescence.TRPM7 channels were inhibited by specific ion channel inhibitor 2-APB.Cell proliferation was assessed by WST-1.Cell apoptosis was evaluated by flow cytometry and Hoechst 33342 staining.【Results】 TRPM7 was expressed in RBL-2H3 observed by Western blot and immunofluorescence,and it was located mainly along the membrane.The application of 2-APB（100μmol/L and 200μmol/L）could significantly inhibit TRPM7 protein expression.The inhibition rates of proliferation by 100μmol/L and 200μmol/L 2-APB were（30.4±4.2）%and（42.0±0.8）%,respectively,while the early apoptosis rates were（36.9±6.7）%and（49.3±1.8）%,and total apoptosis rates were（40.2±7.0）%and（76.8 ±5.5）%,respectively.【Conclusions】TRPM7 channel was expressed in RBL-2H3 and played a role in its proliferation and apoptosis.