Profiling and Bioinformatics Analyses of Differential Circular RNA Expression in Glioblastoma Multiforme Cells Under Hypoxia
Profiling and Bioinformatics Analyses of Differential Circular RNA Expression in Glioblastoma Multiforme Cells Under Hypoxia
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缺氧下多形性胶质母细胞瘤细胞差异性环状RNA表达谱和生物信息学分析
DOI：
10.1007/s12031-022-02090-y
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发表时间：
2022-12-09
影响因子：
3.1
通讯作者：
Chen, Ming
中科院分区：
文献类型：
--
作者：
Chen, Zheng;Su, Shaohua;Chen, Ming
缺氧微环境与GBM的恶性表型高度相关。CircRNA参与GBM的生物学特性，并受HIF-1 α的调控。然而，低氧条件下circRNA在GBM细胞中的差异表达谱和作用仍不清楚。通过circRNA测序分析探讨了低氧条件下LN 229和T98 G中circRNA的表达谱。通过qRT-PCR、RNase R消化反应和桑格测序，选择并验证那些在LN 229和T98 G中均显著失调并且可以在circBase中发现的circRNA。正常细胞系和新鲜GBM组织也用于qRT-PCR验证。通过生物信息学分析评估差异表达的circRNA的作用。在LN 229中有672个失调的circRNA，在T98 G中有698个失调的circRNA。GO分析表明，circRNA表达的改变与GBM细胞的生物发生和代谢有关。KEGG分析表明，TGF-β信号通路、HIF-1信号通路和代谢相关信号通路与低氧条件下差异表达的circRNA密切相关。这些结果通过GSEA分析得到证实。通过qRT-PCR验证LN 229和T98 G中6种选择的和失调的circRNA，包括hsa_circ_0000745、hsa_circ_0020093、hsa_circ_0020094、hsa_circ_0000943、hsa_circ_0004874和hsa_circ_0002359。抑制hsa_circ_0000745可抑制GBM细胞的增殖、迁移和侵袭。以HIF-1 α为中心的circRNA-miRNA-mRNA网络分析表明，6种验证的circRNA与11种相关的miRNA发生了相互作用。低氧条件下GBM细胞circRNA表达失调。这6种circRNA在缺氧条件下可能参与GBM的发生和发展。它们有可能成为GBM的预后标志物和辅助治疗的候选者。
The hypoxia microenvironment is highly associated with GBM's malignant phenotypes. CircRNAs were reported involved in GBM's biological characteristics and regulated by HIF-1 alpha. However, the differential expression profile and role of circRNAs in GBM cells under hypoxia are still unclear. The expression profiles of circRNAs in LN229 and T98G under hypoxia were explored via circRNA sequencing analysis. Those circRNAs significantly dysregulated both in LN229 and T98G and could be found in circBase were selected and validated by qRT-PCR, RNase R digestion reaction, and Sanger sequencing. Normal cell line and fresh GBM tissues were also used for qRT-PCR validation. The roles of differentially expressed circRNAs were evaluated by bioinformatics analyses. There were 672 dysregulated circRNAs in LN229 and 698 dysregulated circRNAs in T98G. GO analysis indicated that the alteration of circRNA expression related to GBM cell's biogenesis and metabolism. KEGG analysis demonstrated that TGF-beta signaling pathway, HIF-1 signaling pathway, and metabolism-related signaling pathway were closely associated with differentially expressed circRNAs under hypoxia. These results were confirmed by GSEA analysis. The 6 selected and dysregulated circRNAs both in LN229 and T98G including hsa_circ_0000745, hsa_circ_0020093, hsa_circ_0020094, hsa_circ_0000943, hsa_circ_0004874, and hsa_circ_0002359 were validated by qRT-PCR. Inhibition of hsa_circ_0000745 inhibited GBM cell's proliferation, migration, and invasion. HIF-1 alpha centered circRNA-miRNA-mRNA networks analysis showed that the 6 validated circRNAs could cross-talk with 11 related miRNAs. The circRNA expressions are dysregulated in GBM cell under hypoxia. The 6 validated circRNAs could participate in GBM's development and progression when hypoxia occurs. They might be the candidates for prognostic markers and adjuvant therapeutics of GBM in the future.