蛋白酶体抑制剂MG132对急性髓系白血病细胞增殖与凋亡的影响

目的探讨蛋白酶体抑制剂MG132对急性髓系白血病(acute myeloid leukemia,AML)细胞增殖与凋亡的影响。方法 qRT-PCR检测9株血液肿瘤细胞株内ADRM1 mRNA表达。利用shRNA干扰HL60细胞内ADRM1表达,将不同浓度的MG132作用于ADRM1干扰前后的HL60细胞24 h,CCK-8法检测各组细胞增殖与活力;Western blot检测各组细胞内ADRM1、UCH37蛋白表达;流式细胞仪分析不同浓度MG132作用下,HL60、NB4细胞凋亡情况。结果血液肿瘤细胞株内ADRM1 mRNA表达上调。成功构建ADRM1 shRNA与scrambled shRNA HL60细胞株。ADRM1基因干扰及给予MG132后,各实验组细胞增殖抑制,活力下降,此时细胞内ADRM1、UCH37蛋白表达下调。随着MG132浓度提高,HL60、NB4细胞凋亡增加;MG132对HL60细胞的促凋亡作用强于NB4细胞。结论血液肿瘤细胞株内ADRM1 mRNA过表达;ADRM1蛋白下调后,UCH37蛋白亦下调,细胞增殖抑制;MG132通过下调ADRM1、UCH37蛋白表达,诱导AML细胞凋亡,抑制细胞增殖与活力;MG132对不同分型AML细胞的促凋亡作用存在个体差异。

Effect of MG132 on acute myeloid leukemia cell proliferation and apoptosis

Aim To investigate the effect of proteasome inhibitor MG132 on the proliferation and apoptosis of acute myeloid leukemia cells.Methods qRT-PCR was used to detect the expression of ADRM1 mRNA in nine blood tumor cell lines.The expression of ADRM1 in HL60 cells was interfered by shRNA;HL60 cells before and after ADRM1 interfered were treated with different MG132 concentrations for 24 h;Then,the cell proliferation and viability were measured with CCK-8 by microplate reader.Meanwhile,the expressions of ADRM1 and UCH37 protein were detected by Western blot.Apoptosis of HL60 and NB4 cells treated with different MG132 concentrations was analyzed by flow cytometry.Results ADRM1 mRNA was up-regulated in blood tumor cell lines.ADRM1 shRNA and scrambled shRNA HL60 cells were successfully constructed.Cell proliferation and viability were inhibited by ADRM1 shRNA interference or decreased with the increase of MG132 concentration;meanwhile,ADRM1 and UCH37 protein expressions were down-regulated.The apoptosis of HL60 and NB4 cells increased with the increase of MG132 concentrations.The apoptotic effect of MG132 on HL60 cells was stronger than that of NB4 cells.Conclusions ADRM1 mRNA is overexpressed in blood tumor cell lines;ADRM1 down-regulation induces UCH37 protein decrease and cell proliferation inhibition.MG132 induces AML cell apoptosis and restrains the proliferation and viability through down-regulating the expression of ADRM1 and UCH37 protein.The apoptotic effect of MG132 on different types of AML cells exists individual differences.