| 盛贤福,王珺,庄海峰,张宇,陈均法.复方苦参注射液影响HL-60细胞迁移的机制研究[J].浙江中西医结合杂志,2019,29(1): |
| 复方苦参注射液影响HL-60细胞迁移的机制研究 |
| Effect of Compound Kushen Injection on the migration of HL-60 cells |
| 投稿时间：2018-04-19 修订日期：2018-07-16 |
| DOI： |
| 中文关键词: 复方苦参注射液 粒细胞集落刺激因子 微小RNA-146a 趋化因子CXC受体4 细胞迁移 |
| 英文关键词:CKI, G-CSF, miR-146a, CXCR4, cell migration |
| 基金项目:浙江省中医药科技计划(编号：2017ZB030) |
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| 中文摘要: |
| 目的：探讨复方苦参注射液（CKI）影响HL-60细胞迁移的机制。方法：选取HL-60细胞株作为研究对象，根据不同加药方案分为阳性对照组粒细胞集落刺激因子（G-CSF ）组（40 ng/mL）、实验组CKI组（0.5 ng/mL）、G-CSF和CKI联合加药组和空白对照组，共培养48 h后采用蛋白印迹（WB）和逆转录聚合酶链反应（RT-PCR）分别检测微小RNA-146a（miR-146a）、趋化因子CXC受体4（CXCR4）等信使RNA（mRNA）、CXCR4蛋白表达变化情况；同时采用Transwell小室检测CKI、G-CSF作用前后细胞迁移率变化情况。结果 ：①与空白对照相比，SDF-1α能促进HL-60细胞的迁移（2.01±0.13 vs 1.05±0.10，P<0.05），而CKI、G-CSF、CKI联合G-CSF均能阻断HL-60细胞向SDF-1α迁移（1.50±0.04、1.68±0.08、1.27±0.02 vs 2.01±0.13，P<0.05）。②与空白对照组（1.00±0.08）相比，CKI、G-CSF处理HL60细胞后，CXCR4的mRNA、蛋白表达均受抑（0.47±0.09、0.75±0.10，P<0.05）；与单独加药组相比，CIK联合G-CSF处理后，CXCR4的mRNA、蛋白表达受抑更明显（0.24±0.04 vs 0.47±0.09、0.75±0.10，P<0.05）。③G-CSF、CKI处理HL-60细胞后，miR-146a表达显著上调（5.59±0.46、3.53±0.39 vs 1.03±0.11，P<0.01）；而联合加药组上调更明显（6.76±0.54 vs 5.59±0.46、3.53±0.39，P<0.05）。结论：CKI能协同G-CSF上调miR-146a表达，通过降低HL-60细胞表面的CXCR4表达而阻断CXCR4/SDF-1α信号轴，最终导致HL-60细胞体外迁移能力明显下降。 |
| 英文摘要: |
| Objective: To explore the mechanism of compound Kushen injection (CKI) affecting the migration of HL-60 cells. Methods: HL-60 cell line was selected as study subject and divided into positive control group (G-CSF group, 40 ng/mL), experimental group (CKI group, 0.5 ng/mL), G-CSF and CKI combination group and blank control group, according to different dosing regimen. Western Blot and RT-PCR were used to detect the expression of miR-146a and CXCR4 mRNA and CXCR4 protein after 48 hours of co-culture. Trans-well chamber was used to detect the changes in cell migration before and after treatment. Results: ①SDF-1α could promote the migration of HL-60 cells （2.01±0.13 vs 1.05±0.10，P<0.05）, but CKI, G-CSF, CKI combined with G-CSF could block the migration of HL-60 cells to SDF-1α（1.50±0.04、1.68±0.08、1.27±0.02 vs 2.01±0.13，P<0.05）. ②Compared with the blank control group（1.00±0.08）, the expression of CXCR4 mRNA and protein were inhibited after treatment of HL60 cells with CKI and G-CSF（0.47±0.09、0.75±0.10，P<0.05）. Compared with the single drug treatment group, the suppression of CXCR4 mRNA and protein expression was more pronounced in the G-CSF and CKI combination group（0.24±0.04 vs 0.47±0.09、0.75±0.10，P<0.05）. ③The expression of miR-146a was significantly up-regulated after HL-60 cells treated with CKI or G-CSF（5.59±0.46、3.53±0.39 vs 1.03±0.11，P<0.01）, while it was significantly increased after combined treatment with HL-60 cells（6.76±0.54 vs 5.59±0.46、3.53±0.39，P<0.05）. Conclusion: CKI can synergize with G-CSF to up-regulate the expression of miR-146a, and block the CXCR4/SDF-1α signaling axis by decreasing the expression of CXCR4 on the surface of HL-60 cells, which ultimately leads to the decrease of the in vitro migration of HL-60 cells. |
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