亚铁螯合酶对人多发性骨髓瘤RPMI8226细胞的影响及作用机制研究

Effects and mechanisms of ferrous chelatase on proliferation of human multiple myeloma RPMl8226 cells

目的探讨亚铁螯合酶(FECH)对人多发性骨髓瘤RPMI8226细胞的影响及作用机制。方法设计并合成靶向FECH的短发夹RNA(shRNA),转染至RPMI8226细胞,并用Western blot法验证。采用CCK-8法测定FECH shRNA RPMI8226细胞和空载对照RPMI8226细胞的增殖能力,流式细胞术检测细胞活性氧(ROS)产生情况,Western blot法检测细胞铁调节蛋白2(IRP2)、转铁蛋白受体1(TfR1)、过氧化氢酶(Catalase)的表达情况。结果与空载对照RPMI8226细胞相比,FECH shRNA RPMI8226细胞增殖能力减弱(P<0.05),ROS产生增多(P<0.05),IRP2、TfR1表达上调(均P<0.05),Catalase表达下调(P<0.05)。结论FECH对维持RPMI8226细胞氧化还原的动态平衡起重要作用。FECH表达受抑制后,RPMI8226细胞IRP2、TfR1表达上调,细胞内铁过载,ROS产生增多,同时抗氧化蛋白Catalases表达下调,细胞抗氧化能力下降,引发细胞氧化应激,抑制细胞增殖。

Objective To investigate the effects of ferrous chelatase(FECH)on proliferation of human multiple myeloma RPMI8226 cells and its mechanism.Methods The shRNA targeting FECH was designed and synthesized,transfected into RPMI8226 cells and confirmed by Western blot.CCK-8 was used to determine the proliferative capacity of FECH shRNA-transfected RPMI8226 cells and control cells.Flow cytometry was used to detect ROS,and Western blot was used to detect the expression changes of TfR1,IRP2 and catalase.Results The proliferation of FECH shRNA-transfected RPMI8226 cells was lower than that of the control cells(P<0.05).The ROS produced by FECH shRNA-transfected RPMI8226 cells were higher than that of the control cells(P<0.05).And the expression of IRP2 and TfR1 in FECH shRNA-transfected RPMI8226 cells were increased(P<0.05),while expression of catalase was decreased(P<0.05).Conclusion FECH may play an important role in maintaining the redox homeostasis of RPMI8226 cells.The inhibited FECH expression can increase the expression of IRP2 and TfR1,overload the intracellular iron,and decrease the anti-oxidation protein Catalase expression,leading to the decrease of cell antioxidant capacity to induce cell oxidative stress and inhibit cell growth ability.