Rolling adhesion of HL-60 cells treated with all-trans retinoic acid on E-selectin

doi:10.3969/j.issn.2095-4344.0499

Abstract

Abstract:

BACKGROUND: All-trans retinoic acid (ATRA) is an ideal therapy for acute promyelocytic leukemia, which can induce promyelocytes to differentiate to mature granulocytes. However, differentiation syndrome is still a high risk for the patients undergoing ATRA therapy. Occurrence of this complication is closely related with cellular morphology change and expression and function of cellular adhesion molecules, especially selectin and integrin families.
OBJECTIVE: To reveal rolling adhesion behavior and mechanical mechanism of ATRA treated HL-60 cells on the substrate coated with E-selectin under different fluid shear forces.
METHODS: Using the equipment of parallel plate flow chamber, untreated and ATRA treated HL-60 cells were driven to roll on E-selectin-coated substrate. The mean rolling velocity and mean stop time were calculated. Here, the HL-60 cells were incubated in the medium containing 1×10^-⁶ mol/L ATRA for 0, 48, 72, 96 hours. The substrates were captured with 40 μg/L E-selectin overnight and the shear stresses were set to 0.02, 0.04, 0.06 Pa.
RESULTS AND CONCLUSION: The velocity of untreated/treated HL-60 cells decreased firstly and then increased with monotonously increasing shear stress. On the contrary, the mean stop time and factional stop time increased firstly and then decreased. Therefore, we deduced that the flow enhanced rolling adhesion was regulated by the catch bond for the HL-60 cells rolling on E-selectin under flow. On the other side, rolling velocities decreased under the same shear stress even if treated with or without ATRA, and the mean stop time and factional stop time increased inversely, which further illustrate the rolling velocity is mainly regulated by stop time.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: HL-60 Cells, E-Selectin, Tretinoin, Cell Adhesion, Tissue Engineering

CLC Number:

R394.2

Xiao Jing, Li Qu-huan, Yang Bi-shan, Fang Ying, Wu Jian-hua. Rolling adhesion of HL-60 cells treated with all-trans retinoic acid on E-selectin[J]. Chinese Journal of Tissue Engineering Research, 2018, 22(13): 2120-2125.

Figures/Tables 2

2.2 滚动速度设置剪切力0.02，0.04，0.06 Pa条件下，经ATRA处理48，72，96 h的HL-60细胞在铺有E-选择素的基质上滚动，观察细胞滚动黏附行为，测定其滚动的平均速度，实验结果见图2。从图可以看出，随着剪切力从0.02 Pa到0.04 Pa的增加，各组细胞滚动速度呈现降低趋势，随着剪切力从0.04 Pa至0.06 Pa的进一步增加，各组细胞滚动速度呈现增加趋势，即随剪切力的单调增加，平均速度曲线呈现先减少后增加的双态性趋势(图2A)。相应地，滚动速度的频率分布亦随剪切力的增加，低滚动速度的细胞所占的比率表现出先多后少的趋势(图2B-D)。而在同一剪切力下，随着药物处理时间的延长，细胞的滚动速度呈现出下降趋势，原因可能在于药物处理使得细胞的黏附性增强。 2.3 瞬时速度为了探索调控平均滚动速度双态性的机制，跟踪每一剪切力和药物处理下的细胞滚动，作出瞬时滚动速度对时间的图形。选择不同剪切力下未经药物处理和药物处理48 h有代表性的细胞各1个作出瞬时滚动速度图。从图3可知，未经药物处理和药物处理的细胞在较低剪切力下，细胞的滚动快速无序，随着剪切力提高至阈值，细胞滚动速度减小而变得逐渐有序，超过阈值之后，随着剪切应力的进一步提高，细胞滚动再次加速并变得无序。平均停留时间表现出规律性先增大后减少的趋势。 2.4 停留时间和停留时间比率为了定量图3所示的滚动行为，进一步使用细胞滚动的二步模型，自编Excel宏，统计细胞的停留时间和停留时间比率，见图4。由图可知，随剪切力的增大，细胞停留时间和停留时间比率亦呈现先增大后减少的趋势，与细胞滚动速度先减少后增大(图3)的趋势互补，这可能说明，滚动速度的减少是由于平均停留时间的增大所致。同样，在相同剪切力条件下，随着药物处理时间的增加，平均停留时间逐渐增大。与随药物处理时间增长平均滚动速度减低的趋势相反，这进一步证明，平均滚动速度和平均停留时间成负相关关系。"

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