长链非编码 RNA HCG9 通过作为 miR-34b-3p 的 ceRNA 促进骨肉瘤的进展。
Long Noncoding RNA HCG9 Promotes Osteosarcoma Progression through RAD51 by Acting as a ceRNA of miR-34b-3p.
机构信息
Department of Orthopedics, The Second Xiangya Hospital, Central South University, Changsha, Hunan 410011, China.
出版信息
Mediators Inflamm. 2021 Aug 18;2021:9978882. doi: 10.1155/2021/9978882. eCollection 2021.
BACKGROUND
Long noncoding RNAs (lncRNAs) have critical regulatory functions in biological and pathological activities during osteosarcoma progression. It is important to elucidate the expression pattern and reveal the underlying mechanisms of the newly identified lncRNAs.
METHODS
Herein, we screened the differentially expressed lncRNAs in osteosarcoma tumors and cell lines using lncRNA microarray. The candidate lncRNA was further verified by qRT-PCR, and the association of gene expression with clinicopathological features was evaluated by Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The targeting miRNA was identified using starBase analysis, and the competing endogenous RNA (ceRNA) network was established by STRING. Overexpression and silence of RNA were detected by qRT-PCR. Osteosarcoma cell proliferation was measured with CCK-8 assay, and the migration and invasion were evaluated with Transwell assay. Colony formation assay was observed. Flow cytometry evaluated the cell cycle. Western blot was performed to detect the mitotic markers and apoptosis-related proteins. A nude mouse tumor formation experiment was used to evaluate osteosarcoma progression . Cooverexpressing miR-34b-3p with RAD51 reversed the miR-34b-3p-induced changes in proliferation, the cell cycle, the expression of H2A.X, and that of apoptosis-related proteins.
RESULTS
HCG9 was identified as osteosarcoma-associated lncRNA. Osteosarcoma tissues and cell lines expressed higher levels of HCG9 as compared to normal tissues and osteoblasts, and high expression of HCG9 was further proved to be related to metastasis and the grade of osteosarcoma in clinical cases. Knockdown of HCG9 inhibited the proliferation, migration, and invasion of osteosarcoma cells. miR-34b-3p was identified as the target of HCG9, and RAD51 acted as a potential target of miR-34b-3p. Cooverexpressing miR-34b-3p with HCG9 partially suppressed the HCG9-stimulated proliferation, migration, and invasion of osteosarcoma cells and delayed the tumor progression .
CONCLUSION
We discovered that lncRNA HCG9 promoted the proliferation of osteosarcoma cells via suppressing miR-34b-3p. Our study provides novel biomarkers and potential therapeutic targets for osteosarcoma treatment.
背景
长链非编码 RNA（lncRNA）在骨肉瘤进展过程中的生物学和病理学活动中具有关键的调节功能。阐明新鉴定的 lncRNA 的表达模式并揭示其潜在机制非常重要。
方法
本研究采用 lncRNA 微阵列筛选骨肉瘤肿瘤和细胞系中的差异表达 lncRNA。通过 qRT-PCR 进一步验证候选 lncRNA，并通过京都基因与基因组百科全书（KEGG）通路分析评估基因表达与临床病理特征的关系。使用 starBase 分析鉴定靶向 miRNA，并通过 STRING 建立竞争内源 RNA（ceRNA）网络。通过 qRT-PCR 检测 RNA 的过表达和沉默。用 CCK-8 测定骨肉瘤细胞增殖，用 Transwell 测定迁移和侵袭，用集落形成实验观察。用流式细胞术评估细胞周期。用 Western blot 检测有丝分裂标记物和凋亡相关蛋白。用裸鼠肿瘤形成实验评估骨肉瘤进展。过表达 miR-34b-3p 并与 RAD51 共转染可逆转 miR-34b-3p 诱导的增殖、细胞周期、H2A.X 表达和凋亡相关蛋白表达的变化。
结果
HCG9 被鉴定为骨肉瘤相关 lncRNA。与正常组织和成骨细胞相比，骨肉瘤组织和细胞系中 HCG9 的表达水平更高，并且 HCG9 的高表达进一步证明与骨肉瘤的转移和分级有关。敲低 HCG9 抑制骨肉瘤细胞的增殖、迁移和侵袭。miR-34b-3p 被鉴定为 HCG9 的靶标，RAD51 是 miR-34b-3p 的潜在靶标。共表达 miR-34b-3p 与 HCG9 部分抑制了 HCG9 刺激的骨肉瘤细胞的增殖、迁移和侵袭，并延缓了肿瘤进展。
结论
我们发现 lncRNA HCG9 通过抑制 miR-34b-3p 促进骨肉瘤细胞的增殖。我们的研究为骨肉瘤治疗提供了新的生物标志物和潜在的治疗靶点。
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