中电导钙激活性钾通道在多发性骨髓瘤细胞增殖、迁移、侵袭和IgE分泌中的作用

Relation of Intermediate-conductance Ca^(2+)-activated K^+ Channels with Ability of Proliferation,Migration,Invasion and IgE Secretion of Multiple Myeloma Cells

本研究旨在探讨中电导钙激活性钾通道(IKCa1)在多发性骨髓瘤细胞增殖、迁移、侵袭及单克隆免疫球蛋白分泌中的作用。应用台盼蓝拒染法检测IKCa1阻滞剂克霉唑(clotrimazole,CLO)对MM细胞株U266生存能力的影响;应用Transwell迁移及Matrigel侵袭实验检测CLO对U266细胞迁移及侵袭能力影响;应用双抗夹心法(ELISA法)检测CLO对U266细胞单克隆免疫球蛋白IgE分泌的影响。结果表明,小剂量CLO(≤1.0μmol/L)对U266细胞活力无明显影响。Transwell迁移实验及Matrigel侵袭实验显示,小剂量CLO(≤1.0μmol/L)作用后,U266迁移及侵袭能力明显降低(P<0.05)。1.00μmol/L CLO作用24 h与48 h后,细胞上清液IgE浓度分别为(4.98±0.39)、(4.38±0.32)ng/ml,24 h与48 h对照组IgE浓度分别为(15.41±1.88)、(21.73±2.01)ng/ml,提示1.00μmol/L组与对照组比较具有显著差异(P<0.05)。结论:CLO通过阻滞IKCa1而抑制MM细胞迁移和侵袭及M蛋白分泌,这为MM靶向治疗提供新思路。

This study was aimed to investigate the effects of the intermediate-conductance Ca2＋ -activated K＋ （IKCal） channels on the proliferation, migration, invasion ability and monoclonal immunoglobulin （IgE） secretion of multiple myeloma （MM） cells. Trypan blue exclusion was used to evaluate the impact of clotrimazole （CLO, an inhibitor of the KCal ） on the survival ability of MM cell line U266 ; transwell chamber and matrigel experiments were used to evaluate the impact of CLO on the ability of the migration and invasion of U266 cells; the influence of CLO on IgE secretion in U266 cells was detected by ELISA. The results showed that small dose of CLO （ ≤ 1.0 μmol/L） could not inhibit the viability of U266 cells. The Transwe11 and Matrigel invasing tests displayed that the cell number moving into lower chamber of transwell decreased after U266 cells treated with small dose of CLO （ ≤ 1.0 μmol/L） . After treating the cells with 1.00 μmol/L CLO for 24 h and 48 h, the concentration of IgE in cell supernatant was （4.98 ± 0. 39） and （4. 38 ± 0. 32） ng/ml, while those in control group were （ 15.41±1.88） and （ 19.73± 2.01 ） ng/ml, respectively, suggesting significant difference between them （ P 〈 0.05 ）. It is concluded that CLO can decrease the ability of migration and monoclonal immunoglobulin secretion of multiple myeloma cells by blocking the IKCal, thus this study provides a new think for the targeted therapy of MM.