High throughput circRNA sequencing analysis reveals novel insights into the mechanism of nitidine chloride against hepatocellular carcinoma
High throughput circRNA sequencing analysis reveals novel insights into the mechanism of nitidine chloride against hepatocellular carcinoma
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高通量circRNA测序分析揭示了氯化两面针碱抗肝细胞癌机制的新见解
DOI：
10.1038/s41419-019-1890-9
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发表时间：
2019-09-10
影响因子：
9
通讯作者：
Luo, Dian-zhong
中科院分区：
文献类型：
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作者：
Xiong, Dan-dan;Feng, Zhen-bo;Luo, Dian-zhong
氯化两面针碱（NC）已被证明对肝细胞癌（HCC）具有抗癌作用。然而，NC对HCC的作用机制仍不清楚。在这项研究中，收集了三对NC处理和NC未处理的HCC异种移植肿瘤组织用于circRNA测序分析。两组间共有297个circRNA表达差异，其中188个上调，109个下调，其中hsa_circ_0088364和hsa_circ_0090049通过实时定量聚合酶链反应进行了验证。体外实验表明，这两种circRNA对肝癌的恶性生物学行为有抑制作用，提示它们可能在肝癌的发生发展中起重要作用。为了阐明这两种circRNA是否在HCC中起“miRNA海绵”的作用，我们分别使用CircInteractome和miRwalk鉴定了circRNA-miRNA和miRNA-mRNA的相互作用。随后，选择857个miRNA相关差异表达基因进行加权基因共表达网络分析。Eigengene turquoise基因组423个基因与肝癌患者的生存时间、病理分级和TNM分期显著相关。基因功能富集分析表明，这423个基因主要在DNA复制和细胞周期相关的生物过程和信号级联中发挥作用。然后鉴定了与HCC患者的临床结果相关的18个hubgenes（SMARD 1、CBX 1、HCFC 1、RBM 12 B、RCC 2、NUP 205、ECT 2、PRIM 2、RBM 28、COPS 7 B、PRRC 2A、GPR 107、ANKRD 52、TUBA 1B、ATXN 7 L3、FUS、MCM 8和RACGAP 1）。这些发现表明hsa_circ_0088364和hsa_circ_0090049及其竞争mRNA之间的串扰可能在HCC中起重要作用，为circRNA作为HCC中NC治疗靶点的潜力提供了有趣的线索。
Nitidine chloride (NC) has been demonstrated to have an anticancer effect in hepatocellular carcinoma (HCC). However, the mechanism of action of NC against HCC remains largely unclear. In this study, three pairs of NC-treated and NC-untreated HCC xenograft tumour tissues were collected for circRNA sequencing analysis. In total, 297 circRNAs were differently expressed between the two groups, with 188 upregulated and 109 downregulated, among which hsa_circ_0088364 and hsa_circ_0090049 were validated by real-time quantitative polymerase chain reaction. The in vitro experiments showed that the two circRNAs inhibited the malignant biological behaviour of HCC, suggesting that they may play important roles in the development of HCC. To elucidate whether the two circRNAs function as “miRNA sponges” in HCC, we identified circRNA-miRNA and miRNA-mRNA interactions by using the CircInteractome and miRwalk, respectively. Subsequently, 857 miRNA-associated differently expressed genes in HCC were selected for weighted gene co-expression network analysis. Module Eigengene turquoise with 423 genes was found to be significantly related to the survival time, pathology grade and TNM stage of HCC patients. Gene functional enrichment analysis showed that the 423 genes mainly functioned in DNA replication- and cell cycle-related biological processes and signalling cascades. Eighteen hubgenes (SMARCD1, CBX1, HCFC1, RBM12B, RCC2, NUP205, ECT2, PRIM2, RBM28, COPS7B, PRRC2A, GPR107, ANKRD52, TUBA1B, ATXN7L3, FUS, MCM8 and RACGAP1) associated with clinical outcomes of HCC patients were then identified. These findings showed that the crosstalk between hsa_circ_0088364 and hsa_circ_0090049 and their competing mRNAs may play important roles in HCC, providing interesting clues into the potential of circRNAs as therapeutic targets of NC in HCC.