摘要
最近研究表明,DJ-1在许多肿瘤中过表达,而且DJ-1的核表达与肿瘤的生物学行为有关.本文主要研究二烯丙基二硫(DADS)对DJ-1核定位高表达人白血病HL-60细胞生物行为学的影响,阐明DJ-1在细胞核内的功能,为临床诊断及治疗过程提供一个潜在的治疗靶点.通过基因转染技术建立核内高表达HL-60细胞株(DJ-1/HL-60),利用软琼脂集落形成实验、 MTT法、间接免疫荧光细胞化学实验、硝基蓝四氮唑(NBT)还原比色实验评估HL-60细胞的增殖与分化,DJ-1核定位过表达促进HL-60细胞的增殖并抑制其分化. Transwell迁移侵袭小室实验表明DJ-1核定位过表达可以促进HL-60的迁移和侵袭能力. Western blot结果表明DADS具有抑制HL-60细胞中核内DJ-1蛋白表达的能力.说明DJ-1核定位高表达具有促进HL-60细胞增殖和迁移侵袭及抑制HL-60细胞分化的作用,DADS可以诱导DJ-1核定位高表达HL-60细胞分化及抑制迁移侵袭, DJ-1核定位高表达可减弱DADS抑制HL-60细胞增殖的作用.
Recent studies have revealed that DJ-1 is overexpressed in multiple forms of human tumors, and nuclear overexpression of DJ-1 is correlated with the biological behavior of tumor. The purpose of the present study was to test for possible DADS effects on Human Leukemia HL-60 cells which DJ-1 is overexpressed, and to clarify the function of DJ-1 in the nucleus.Gene transfection technology was used to establish stable HL-60 cell lines with nuclear overexpression of DJ-1(DJ-1/HL-60). Soft agar colony formation, MTT, NBT and indirect immunofluorescence were used to evaluate the proliferation, differentiation,migration and invasion of HL-60 cells. The nuclear overexpression of DJ-1 protein may promote the proliferation and weaken the differentiation of HL-60 cells. Transwell migration and invasion assays showed that the nuclear localization and overexpression of DJ-1 can promote the migration and invasion capability of HL-60. Western blot analysis revealed that the expression of DJ-1 was inhibited in HL-60 cells with nuclear overexpression of DJ-1 after treatment with DADS. These results defined the impacts of DJ-1 on biological behavior of the Human Leukemia HL-60 cells by DADS, the proliferation, migration and invasion were inhibited and the differentiation was induced in HL-60 cells with nuclear overexpression of DJ-1, after treatment with DADS.
引文
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