miR-143通过抑制AF9调控SKM-1细胞增殖和凋亡

崔佳奇; 魏春梅; 邓琳丽; 王利

doi:10.16016/j.1000-5404.201804133

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miR-143通过抑制AF9调控SKM-1细胞增殖和凋亡

基础医学 | 更新时间：2026-01-09

- miR-143通过抑制AF9调控SKM-1细胞增殖和凋亡
- MiR-143 regulates proliferation and apoptosis of SKM-1 cells in vitro by inhibiting AF9
- 第三军医大学学报 2018年40卷第20期页码：1826-1832
- 作者机构：
  
  400016 重庆，重庆医科大学附属第一医院血液内科
- 作者简介：
  
  [王利，E-mail: liwangls@yahoo.com WANG Li, E-mail: liwangls@yahoo.com
- 基金信息：
  
  国家自然科学基金面上项目(81250034);重庆市教委资助课题(KJ1702017)
- DOI：10.16016/j.1000-5404.201804133
  中图分类号： R394-33;R394.12;R551.1
- 收稿：2018-04-19，
  
  修回：2018-07-13，
  
  纸质出版：2018-10-30
- 稿件说明：
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崔佳奇, 魏春梅, 邓琳丽, 等. miR-143通过抑制AF9调控SKM-1细胞增殖和凋亡[J]. 第三军医大学学报, 2018,40(20):1826-1832.

Jiaqi CUI, Chunmei WEI, Linli DENG, et al. MiR-143 regulates proliferation and apoptosis of SKM-1 cells in vitro by inhibiting AF9[J]. Journal of Third Military Medical University, 2018, 40(20): 1826-1832.
崔佳奇, 魏春梅, 邓琳丽, 等. miR-143通过抑制AF9调控SKM-1细胞增殖和凋亡[J]. 第三军医大学学报, 2018,40(20):1826-1832. DOI： 10.16016/j.1000-5404.201804133.

Jiaqi CUI, Chunmei WEI, Linli DENG, et al. MiR-143 regulates proliferation and apoptosis of SKM-1 cells in vitro by inhibiting AF9[J]. Journal of Third Military Medical University, 2018, 40(20): 1826-1832. DOI： 10.16016/j.1000-5404.201804133.

摘要

目的

探讨miR-143在骨髓增生异常综合征(myelodysplastic syndrome，MDS)中的表达情况及AF9对人骨髓增生异常综合征细胞株SKM-1增殖和凋亡的影响。

方法

利用实时荧光定量逆转录PCR(qRT-PCR)检测收集的33位MDS/AML患者以及11位健康人(健康对照)的骨髓标本中miR-143的表达；细胞实验分为过表达组(SKM-1细胞感染含LV-hsa-miR-143的慢病毒载体)、低表达组(SKM-1细胞感染含LV-hsa-miR-143-inhibitors的慢病毒载体)和阴性对照组(SKM-1细胞感染含空病毒LV-control的慢病毒载体)，分别采用CCK-8法、流式细胞仪检测各组SKM-1细胞增殖、周期和凋亡情况；双荧光素酶报告基因检测miR-143对下游靶基因AF9在转录水平的影响；Western blot测定miR-143对下游靶基因AF9蛋白表达水平的影响。

结果

与健康对照比较，MDS患者的miR-143相对表达水平明显降低(

0.05)；过表达组的光密度值在第3天为(0.562±0.069)，明显低于阴性对照组(

0.05)；低表达组的光密度值在第3天为(1.537±0.094)，明显高于阴性对照组(

0.05)；过表达组G

期细胞比例为(57.92±3.58)%，明显高于阴性对照组(

0.05)；低表达组G

期细胞比例为(29.73±2.44)%，明显低于阴性对照组(

0.05)；过表达组细胞凋亡率为(29.80±2.53)%，明显高于阴性对照组(

0.05)；而低表达组细胞凋亡率为(5.17±1.04)%，明显低于阴性对照组(

0.05)；miR-143可以显著抑制AF9 3’-UTR野生型的报告基因活性；低表达组AF9蛋白表达水平显著高于阴性对照组(

0.05)。

结论

miR-143在MDS患者中的相对表达水平明显减低；过表达miR-143后骨髓增生异常综合征SKM-1细胞增殖活性降低、G

期细胞阻滞增多、凋亡率增加；同时miR-143能够负性调节其靶基因AF9。

Abstract

Objective

To investigate the expression of miR-143 in the bone marrow of patients with myelodysplastic syndrome (MDS) and whether it inhibits SKM-1 cell proliferation and apoptosis by inhibiting AF9.

Methods

Real-time quantitative RT-PCR (qRT-PCR) was used to detect the expression of miR-143 in bone marrow samples from 33 patients with MDS/acute myeloid leukemia (AML) and 11 healthy individuals. In the

in vitro

experiment

SKM-1 cell line

a cell model of MDS

was infected with the lentiviral vectors LV-hsa-miR-143

LV-hsa-miR-143-inhibitors

or LV-control

and the changes in the cell proliferation

cell cycle and apoptosis were detected using CCK-8 assay and flow cytometry. Dual-luciferase reporter assay was used to assess the effect of miR-143 expression modulation on the transcriptional level of the downstream target gene AF9; the expression of AF9 protein was detected using Western blotting.

Results

Compared with the healthy individuals

MDS patients showed a significantly lowered expression level of miR-143 in the bone marrow. In SKM-1 cells

mi-R143 overexpression significantly suppressed cell proliferation

and miR-143 knockdown significantly inhibited cell proliferation as compared with the control cells (

0.05); mi-R143 overexpression significantly increased the percentage of cells in G

phase compared with the negative control cells [(57.92±3.58)%

(38.75±2.78)%

0.05

]

and knockdown of miR-143 expression obviously lowered the percentage of cells in G

phase [(29.73±2.44)%

0.05

]

; mi-R143 overexpression significantly increased the cells apoptosis rate to (29.80±2.53)% (

0.05)

and miR-143 knockdown decreased the cell apoptosis rate to (5.17±1.04)% (

0.05)

as compared to the rate of (13.92±1.49)% in the negative control cells. Overexpression of miR-143 significantly inhibited the activity of wild-type AF9 3'-UTR reporter gene

and compared with the control cells

the cells with miR-143 knockdown showed significantly increa

sed expression level of AF9 protein (

0.05).

Conclusions

The patients with MDS show obviously lowered expression level of miR-143 in the bone marrow. Overexpression of miR-143 suppresses the proliferation

induces cell cycle arrest in G

phase and promotes apoptosis of SKM-1 cells by negatively regulating its target gene AF9.

关键词

Keywords

references

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