The Effects Of DADS On Biological Behaviour Of The Human Leukemia HL-60Cells With Nuclear Localization Of DJ-1Overexpression

Objective: To study biological behaviour effects of DADS on the human leukemiaHL-60cells with nuclear localization overexpression of DJ-1, and to explore thefunction of DJ-1in the nucleus, it provides a potential biomarkers or effectivemedicines for tumors in the process of clinical diagnosis and treatment.Methods：The recombined plasmid pCMV/myc/nuc-EGFP-DJ-1was transfected intoHL-60cell by lipo fectamine2000, and G418was used to screen out the positiveclone. The localization expression of DJ-1in HL-60cells were verified by fluorescentmicroscope and Western blot. After the success of the transfection experiment isdivided into three groups: the HL-60, Vector and DJ-1/HL-60. The effect of thenucleus overexpression of DJ-1protein on proliferation, cycle, migration,differentiation and DJ-1protein expression whit DADS treatment in HL-60cells weredetected by soft agar colony formation, MTT, flow cytometry, Transwell invasionchamber assays, NBT, Giemsa, indirect immunofluorescence test and Western blot.Results：1.The successful construction of HL-60cells with stably nuclear localizationoverexpression of DJ-1Sequencing results showed that plasmid pCMV/myc/nuc-EGFP-DJ-1is correct.Fluorescent microscope and Western blot showed that DJ-1protein was stablyincreased in the HL-60cell nuclear, and suggested that HL-60cells with stablynuclear localization overexpression of DJ-1was successfully constructed.2.The effects of DADS on biological behaviour of the human leukemia HL-60cellswith nuclear localization overexpression of DJ-1Soft agar colony formation, MTT, flow cytometry, Transwell invasion chamberassays and Western blot showed that the nucleus overexpression of DJ-1protein promoted the proliferation, migration and invasion capability of HL-60cells withnuclear localization overexpression of DJ-1and induced S/G2arres（tP<0.05）. DADSinhibited the proliferation, migration and invasion in three groups of cells and inducedG0/G1arrest （P<0.05）. These results showed that the DADS inhibited theproliferation, migration and invasion in HL-60cells with nuclear localizationoverexpression of DJ-1.NBT, Giemsa and indirect immunofluorescence test showed that thedifferentiation capability of HL-60cells with nuclear localization overexpression ofDJ-1were significantly decreased （P<0.05）, suggesting that the nucleusoverexpression of DJ-1protein inhibited the differentiation in HL-60cells. DADSinhibited the differentiation capability of HL-60cells with nuclear localizationoverexpression of DJ-1（P<0.05）.Conclusion：1.The nucleus overexpression of DJ-1protein may promote the proliferation,migration, invasion and weaken the differentiation of HL-60cells.2.DADS can induce the differentiation and reduce the invasion of HL-60cells withnuclear localization overexpression of DJ-13.The nucleus overexpression of DJ-1protein can decrease the proliferation of HL-60by DADS.