DRAM-1（自噬调控基因-1）慢病毒包装及对 HL60细胞分化作用的研究

摘要

Objective The aim of our test is to observe the changes of DRAM-1 in differentiated HL60 cells,and to observe the inhibitory effect of DRAM-1 expression on the differentiation of HL60 cell line. Methods We mixed retinoic acid 1 uMand the amount of culture medium into 0.2 ×1 06 /ml HL60 cell line,then we used flow cytometry to detect the expression of cell surface CD1 1 in 24h,48h,72h,96h respectively and used RT-PCR to detect the expression of DRAM-1 mRNA,CEB-PE,CSF3R.We used lentiviral transfection of 293T cells,and collected the culture supernatant to infect HL60 cells,then used to detect the expression of lentivirus infection after DRAM-1 mRNA. Results ATRA treatment group compared with the un-treated group,DRAM-1 mRNA expression was significantly increased in the process of differentiation in HL60 cell.After lentivi-ral infected cells,RT-PCR showed that the expression of DRAM-1 was lower than control group,with the same ATRA,the ex-pression of CD1 1 b decreased. Conclusion The expression of DRAM-1 was increased in the differentiation of HL60 cell lines.Inhibition of the expression of DRAM-1 blocked differentiation of HL60 cell line.%目的：观察 HL60细胞分化过程中 DRAM-1变化，观察抑制 DRAM-1表达对 HL60细胞系分化的影响。方法0.2×106/ml 的 HL60细胞系中加入维甲酸1μmol，分别在24h、48h、72h、96h 流式细胞仪检测细胞表面CD11 b 表达，RT-PCR 检测 DRAM-1 mRNA、CEBPE、CSF3R 表达。采用慢病毒转染293T 细胞，收集培养液上清，感染 HL60细胞，RT-PCR 检测慢病毒感染后的 DRAM-1 mRNA 表达。结果ATRA 处理组较未处理组 DRAM-1 mRNA 在HL60细胞系分化过程中表达明显增高。慢病毒感染细胞后，RT-PCR 显示 DRAM-1表达较对照组降低，等量 ATRA 处理，CD11 b 表达下降。结论DRAM-1在 HL60细胞系分化过程中表达增高。抑制 DRAM-1表达阻碍 HL60细胞系分化。