Down-regulating HOXA-AS2/miR-20a pathway promotes apoptosis and inhibits invasion and migration of skin squamous carcinoma cells

Wen XU; Xuhong LIU; Hua LIU

doi:10.16016/j.1000-5404.202005261

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Down-regulating HOXA-AS2/miR-20a pathway promotes apoptosis and inhibits invasion and migration of skin squamous carcinoma cells

Basic Medicine | 更新时间：2026-01-09

- Down-regulating HOXA-AS2/miR-20a pathway promotes apoptosis and inhibits invasion and migration of skin squamous carcinoma cells
- Journal of Third Military Medical University Vol. 42, Issue 20, Pages: 2003-2011(2020)
- 作者机构：
  
  1.225003 江苏扬州，武警江苏省总队医院:皮肤科
  2.225003 江苏扬州，武警江苏省总队医院:门诊部
  3.225003 江苏扬州，武警江苏省总队医院:特色专业一科
- 作者简介：
- 基金信息：
- DOI：10.16016/j.1000-5404.202005261
  CLC： R394.3;R730.23;R739.5
- Received：28 May 2020，
  
  Revised：2020-08-12，
  
  Published：30 October 2020
- 稿件说明：
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Wen XU, Xuhong LIU, Hua LIU. Down-regulating HOXA-AS2/miR-20a pathway promotes apoptosis and inhibits invasion and migration of skin squamous carcinoma cells[J]. Journal of Third Military Medical University, 2020, 42(20): 2003-2011.
DOI：

Wen XU, Xuhong LIU, Hua LIU. Down-regulating HOXA-AS2/miR-20a pathway promotes apoptosis and inhibits invasion and migration of skin squamous carcinoma cells[J]. Journal of Third Military Medical University, 2020, 42(20): 2003-2011. DOI： 10.16016/j.1000-5404.202005261.

摘要

目的

研究下调长链非编码RNA(lncRNA) HOXA-AS2/miR-20a通路对皮肤鳞癌细胞凋亡和侵袭的影响。

方法

qRT-PCR检测皮肤鳞癌组织中HOXA-AS2的表达变化。在皮肤鳞癌细胞A431中转染HOXA-AS2 siRNA，MTT测定细胞增殖，克隆形成实验测定细胞克隆形成能力，流式细胞仪测定细胞凋亡，Transwell小室实验检测细胞迁移和侵袭，采用Western blot检测MMP-2、MMP-9、C-Caspase-3蛋白表达。生物信息学软件预测HOXA-AS2和miR-20a有互补结合位点，荧光素酶报告系统鉴定二者的靶向关系。qRT-PCR检测皮肤鳞癌组织中miR-20a的表达变化。在皮肤鳞癌细胞A431中共转染HOXA-AS2 siRNA、miR-20a inhibitor，检测细胞增殖、克隆、凋亡、迁移、侵袭能力的变化。

结果

HOXA-AS2在皮肤鳞癌组织中的表达升高。转染HOXA-AS2 siRNA后的皮肤鳞癌细胞A431增殖、克隆、侵袭、迁移能力下降，细胞凋亡水平升高，MMP-2、MMP-9蛋白表达减少，C-Caspase-3蛋白表达增高。下调HOXA-AS2靶向促进miR-20a表达。miR-20a在皮肤鳞癌组织中的表达与HOXA-AS2负相关。miR-20a inhibitor可以逆转HOXA-AS2 siRNA对皮肤鳞癌细胞A431增殖、克隆、侵袭、迁移和凋亡作用。

结论

下调HOXA-AS2/miR-20a通路降低皮肤鳞癌细胞增殖、克隆、侵袭、迁移能力，诱导细胞凋亡。

Abstract

Objective

To study the effect of down-regulation of long non-coding RNA (lncRNA) HOXA cluster antisense RNA 2 (HOXA-AS2)/miR-20a pathway on apoptosis and invasion of skin squamous cell carcinoma cells

in vitro

Methods

qRT-PCR was used to detect the expression of HOXA-AS2 and miR-20a in 50 clinical specimens of skin squamous cell carcinoma and paired adjacent tissues. Skin squamous carcinoma cell line A431 was transfected with HOXA-AS2 siRNA

and the changes in cell proliferation

clonogenic ability

apoptosis

migration and invasion were evaluated by using MTT assay

clone formation experiment

flow cytometry

and Transwell assay respectively. Western blotting was performed to detect the expression of MMP-2

MMP-9

and cleaved caspase-3 proteins in the cells. Bioinformatics software predicted that HOXA-AS2 and miR-20a had complementary binding sites

and a luciferase reporter system was employed to verify their targeting relationship. A431 cells were co-transfected with HOXA-AS2 siRNA and miR-20a inhibitor

and the changes in cell proliferation

clone formation

apoptosis

migration and invasion were assessed.

Results

The expression level of HOXA-AS2 was increased in skin squamous cell carcinoma tissues. Transfection of A431 cells with HOXA-AS2 siRNA significantly suppressed the cell proliferation

clone formation

invasion and migration

promoted cell apoptosis

decreased the expression of MMP-2 and MMP-9 proteins

and increased the expression of cleaved caspase-3 protein. Down-regulation of HOXA-AS2 significantly up-regulated miR-20a expression. The expression level of miR-20a in skin squamous cell carcinoma tissues was negatively correlated with that of HOXA-AS2. In A431 cells

miR-20a inhibitor obviously reversed the effects of HOXA-AS2 siRNA on proliferation

clone formation

invasion

migration and apoptosis of the cells.

Conclusion

Down-regulation of HOXA-AS2/miR-20a pathway can promote the expression of miR-20a

suppress the proliferation

clone formation

invasion and migration

and induce apoptosis of skin squamous cell carcinoma cells

in vitro

关键词

Keywords

references

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