HOTAIR 通过靶向 miR-219 抑制脑胶质母细胞瘤细胞的增殖。
HOTAIR inhibits the proliferation of glioblastoma cells by targeting miR-219.
机构信息
Department of Diagnostic, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
Department of Neurosurgery, Yunnan First People's Hospital, Kunming, Yunnan, China.
出版信息
Cancer Biomark. 2020;28(1):41-47. doi: 10.3233/CBM-190467.
BACKGROUND
To investigate the influences of HOX transcript antisense ribonucleic acid (HOTAIR) on the proliferation and apoptosis of glioblastoma cells by targeting micro RNA (miR)-219.
OBJECTIVE
With glioblastoma cell line U87 as the object, the changes in expression levels of HOTAIR and miR-219 in each group were detected via quantitative real-time polymerase chain reaction (qRT-PCR) after HOTAIR in U87 cell lines was knocked down using small interfering RNA (siRNA). Then the cell proliferation in each group was determined using Cell Counting Kit-8 (CCK-8) and colony formation assays. Flow cytometry was applied to detect the cell apoptosis, and Western blotting assay was adopted to measure the changes in protein levels of Cyclin D1 and Bcl-2-associated X protein (Bax). After miR-219 knockdown with siRNA, the changes in expression levels of HOTAIR and miR-219 in each group were examined through qRT-PCR, and the cell proliferation was tested by CCK-8 assay.
RESULTS
After interference inHOTAIR using siRNA, compared with those in control group, the RNA expression level of HOTAIR was decreased remarkably (p< 0.05), the RNA expression level of miR-219 was increased notably (p< 0.05), the cell proliferation rate was inhibited evidently (p< 0.05), the apoptosis rate was enhanced obviously (p< 0.05), the protein expression level of Cyclin D1 declined markedly (p< 0.05), and the protein expression level of Bax rose distinctly (p< 0.05) in HOTAIR-siRNA group. After miR-219 knockdown with siRNA, the cell proliferation rate was raised remarkably (p< 0.05), but there was no significant change in the RNA expression level of HOTAIR (p> 0.05).
CONCLUSION
HOTAIR can repress the proliferation and promote the apoptosis of glioblastoma cells by targeting miR-219.
背景
通过靶向 microRNA（miR）-219 来研究 HOX 转录反义 RNA（HOTAIR）对神经胶质瘤细胞增殖和凋亡的影响。
目的
以神经胶质瘤细胞系 U87 为研究对象，采用小干扰 RNA（siRNA）敲低 U87 细胞系中的 HOTAIR，通过实时定量聚合酶链反应（qRT-PCR）检测各组 HOTAIR 和 miR-219 的表达水平变化。然后用细胞计数试剂盒-8（CCK-8）和集落形成实验测定各组细胞增殖情况。采用流式细胞术检测细胞凋亡，采用 Western blot 检测细胞周期蛋白 D1（Cyclin D1）和 Bcl-2 相关 X 蛋白（Bax）的蛋白水平变化。用 siRNA 敲低 miR-219 后，通过 qRT-PCR 检测各组 HOTAIR 和 miR-219 的表达水平变化，用 CCK-8 实验检测细胞增殖情况。
结果
用 siRNA 干扰 HOTAIR 后，与对照组相比，HOTAIR 的 RNA 表达水平显著降低（p<0.05），miR-219 的 RNA 表达水平明显升高（p<0.05），细胞增殖率明显受到抑制（p<0.05），细胞凋亡率明显升高（p<0.05），Cyclin D1 的蛋白表达水平明显下降（p<0.05），Bax 的蛋白表达水平明显升高（p<0.05）。用 siRNA 敲低 miR-219 后，细胞增殖率明显升高（p<0.05），但 HOTAIR 的 RNA 表达水平没有明显变化（p>0.05）。
结论
HOTAIR 可通过靶向 miR-219 抑制神经胶质瘤细胞的增殖，促进其凋亡。
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