TRPM7通道对人口腔颊黏膜成纤维细胞增殖和迁移的影响

Effect of transient receptor potential melastatin 7 channel on the proliferation and migration of human buccal mucosal fibroblasts

目的:探讨瞬时受体电位离子通道TRPM7(transient receptor potential melastatin-7)在人口腔颊黏膜成纤维细胞增殖和迁移中的作用。方法:逆转录聚合酶链式反应、免疫荧光法及全细胞膜片钳检测成纤维细胞中TRPM7的表达。MTT和细胞划痕实验分别检测应用2-氨基乙基联苯基硼酸酯(2-APB)或shRNA抑制TRPM7通道对成纤维细胞增殖和迁移的影响。结果:人口腔颊黏膜成纤维细胞功能性表达TRPM7通道。TRPM7 shRNA作用48 h后成功阻断TRPM7通道,500μmol/L2-APB可抑制其表达,两组与对照组相比,膜片钳技术记录到TRPM7样电流明显减弱,在24 h后均明显抑制成纤维细胞的增殖和迁移能力,差异有统计学意义(P<0.05)。结论:TRPM7通道参与人口腔颊黏膜成纤维细胞增殖以及迁移能力的调控。

Objective:To investigate the role of transient receptor potential melastatin 7(TRPM7)in the prolifer-ation and migration of human buccal mucosal fibroblasts.Methods:Reverse transcription-polymerase chain reac-tion,immunofluorescence method,and whole-cell patch clamp were used to determine the expression of TRPM7 infibroblasts.MTT and wound-healing assays were used to examine the effects of inhibiting TRPM7 channels using2-aminoethyl diphenylborinate(2-APB)or shRNA on fibroblast proliferation and migration.Results:TRPM7 chan-nels were functionally expressed in human buccal mucosal fibroblasts.TRPM7 channels were successfully blockedafter 48 h of TRPM7 shRNA treatment,and their expression was inhibited by 500μmol/L of 2-APB.Comparedwith the control group,the TRPM7-like currents were significantly decreased in the two experimental groups accord-ing to patch-clamp recording,and the proliferation and migration abilities of fibroblasts were significantly sup-pressed after 24 h of treatments(P<0.05).Conclusion:The TRPM7 channel is involved in the regulation of prolif-eration and migration of human buccal mucosal fibroblasts.