CAJ | 학술논문

目的体外探讨A23187快速将人慢性白血病K562细胞定向诱导分化为树突细胞(DC)的实验方法.方法 K562细胞在含有A23187或细胞因子的条件下诱导分化为DC,倒置显微镜下观察细胞形态的变化,流式细胞术检测DC表面标志的改变,逆转录-聚合酶链反应(RT-PCR)检测各表面标志mRNA水平的变化,四甲基偶氮唑蓝(MTT)比色法检测其刺激淋巴细胞增殖的能力.结果 A23187以385 ng/ml的浓度诱导4 d后,倒置显微镜下可见部分K562细胞形态具有典型的树突样特征,DC标志CD1a、CD83、人类白细胞抗原(HLA)-DR、CD86、CD80在A23187组的表达较阴性对照组均有明显升高,其中A23187组分别为6.65±2.70、7.37±2.40、6.24±4.29、21.60±3.84、18.52±4.48,阴性对照组分别为2.80±0.52、1.85±0.56、2.25±0.47、6.69±0.83、9.%±3.53,差异均有统计学意义(P<0.05).K562细胞来源的DC具有刺激淋巴细胞增殖的能力.结论 A23187可以快速诱导人白血病细胞分化为有活性的DC.
목적 체외탐토A23187쾌속장인만성백혈병K562세포정향유도분화위수돌세포(DC)적실험방법.방법 K562세포재함유A23187혹세포인자적조건하유도분화위DC,도치현미경하관찰세포형태적변화,류식세포술검측DC표면표지적개변,역전록-취합매련반응(RT-PCR)검측각표면표지mRNA수평적변화,사갑기우담서람(MTT)비색법검측기자격림파세포증식적능력.결과 A23187이385 ng/ml적농도유도4 d후,도치현미경하가견부분K562세포형태구유전형적수돌양특정,DC표지CD1a、CD83、인류백세포항원(HLA)-DR、CD86、CD80재A23187조적표체교음성대조조균유명현승고,기중A23187조분별위6.65±2.70、7.37±2.40、6.24±4.29、21.60±3.84、18.52±4.48,음성대조조분별위2.80±0.52、1.85±0.56、2.25±0.47、6.69±0.83、9.%±3.53,차이균유통계학의의(P<0.05).K562세포래원적DC구유자격림파세포증식적능력.결론 A23187가이쾌속유도인백혈병세포분화위유활성적DC.
Objective To explore the method for differentiation induction of leukemia cells into dendritic cells(DC) by A23187 in vitro. Methods Chronic myeloid leukemia K562 cells were cultured with A23187 or cytokine to induce differentiation and form DC. The morphologic features of cells were observed under inverted microscope, the changes of DC surface marks were determined by flow cytometry and RT-PCR, the ability to stimulate lymphocyte proliferation was tested by MTT colorimetry. Results Under the condition of the does (385 ng/ml) of A23187 for four days, some of K562 cells were found in typical dendritic appearance.The expression of DC markers CD1a,CD83 ,HLA-DR,CD86 and CD80 was 6.65 ±2.70,7.37 ±2.40,6.24 ±4.29, 21.60 ± 3.84, 18.52 ± 4.48 repectively, and increased obviously compared with the negative control group(2.80 ±0.52,1.85 ±0.56,2.25 ±0.47,6.69 ±0.83,9.96 ±3.53). The differences had statistical significance (P < 0.05). K562 cells derived from DC acquired the ability to stimulate lymphocyte proliferation.Conclusion A23187 can induce the leukemia cells differenntiation into activated DC-like cells rapidly.