HuR基因沉默通过与长链非编码RNA XIST结合的AGO2抑制骨肉瘤细胞上皮-间质转化

Silencing of HuR Inhibits Osteosarcoma Cell Epithelial-Mesenchymal Transition AGO2 in Association With Long Non-Coding RNA XIST.

作者信息

Liu Yongming, Zhang Yuan, Zhang Jinxue, Ma Jingchang, Xu Xuexue, Wang Yuling, Zhou Ziqing, Jiang Dongxu, Shen Shen, Ding Yong, Zhou Yong, Zhuang Ran

机构信息

Orthopedic Department of Tangdu Hospital, The Fourth Military Medical University, Xi'an, China.

Institute of Medical Research, Northwestern Polytechnical University, Xi'an, China.

出版信息

Front Oncol. 2021 Mar 19;11:601982. doi: 10.3389/fonc.2021.601982. eCollection 2021.

BACKGROUND

Osteosarcoma (OS) is a highly malignant and aggressive bone tumor. This study was performed to explore the mechanisms of HuR (human antigen R) in the progression of OS.

METHODS

HuR expression levels in OS tissues and cells were detected by immunohistochemistry and western blotting. HuR siRNA was transfected into SJSA-1 OS cells to downregulate HuR expression, and then cell proliferation, migration, and epithelial-mesenchymal transition (EMT) were evaluated. RNA immunoprecipitation was performed to determine the association of the long non-coding RNA (lncRNA) XIST and argonaute RISC catalytic component (AGO) 2 with HuR. Fluorescence hybridization analysis was performed to detect the expression of lncRNA XIST. Western blotting and immunofluorescence assays were performed to observe AGO2 expression after HuR or/and lncRNA XIST knockdown.

RESULTS

Knockdown of HuR repressed OS cell migration and EMT. AGO2 was identified as a target of HuR and silencing of HuR decreased AGO2 expression. The lncRNA XIST was associated with HuR-mediated AGO2 suppression. Moreover, knockdown of AGO2 significantly inhibited cell proliferation, migration, and EMT in OS.

CONCLUSION

Our findings indicate that HuR knockdown suppresses OS cell EMT by regulating lncRNA XIST/AGO2 signaling.

背景

骨肉瘤（OS）是一种高度恶性且侵袭性强的骨肿瘤。本研究旨在探讨人抗原R（HuR）在骨肉瘤进展中的作用机制。

方法

采用免疫组织化学和蛋白质印迹法检测骨肉瘤组织和细胞中HuR的表达水平。将HuR小干扰RNA（siRNA）转染至SJSA-1骨肉瘤细胞中以下调HuR表达，随后评估细胞增殖、迁移及上皮-间质转化（EMT）情况。进行RNA免疫沉淀以确定长链非编码RNA（lncRNA）XIST和AGO2（精氨酸酶RISC催化组分2）与HuR的关联。进行荧光杂交分析以检测lncRNA XIST的表达。在敲低HuR或/和lncRNA XIST后，采用蛋白质印迹法和免疫荧光分析观察AGO2的表达情况。