SNHG9 promotes Hepatoblastoma Tumorigenesis via miR-23a-5p/Wnt3a Axis
SNHG9 promotes Hepatoblastoma Tumorigenesis via miR-23a-5p/Wnt3a Axis
复制标题
SNHG9 通过 miR-23a-5p/Wnt3a 轴促进肝母细胞瘤肿瘤发生
DOI：
10.21203/rs.3.rs-335750/v1
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发表时间：
2021
影响因子：
3.9
通讯作者：
ari
中科院分区：
文献类型：
--
作者：
Sun Gui Feng;Rajeev Bh;ari;Liu Ya;Bian Zhixuan;Pan Quihui;Zhu Jiabei;Mao Sewi;Zhen Ni;Wang Jing;Ma Ji;Ramesh Bh;ari
背景：肝母细胞瘤是一种常见于0-5岁儿童的肝脏肿瘤。越来越多的研究表明lncRNA在不同的癌症进展和发展中的潜在作用，包括肝母细胞瘤。SnoRNA宿主基因9（SNHG 9）与不同人类癌症的进展相关，但其在肝母细胞瘤中的特定分子机制并不未知。研究方法：在这项研究中，我们估计SNHG 9在肝母细胞瘤组织和细胞系的定量实时聚合酶链反应（qRT-PCR）的表达。接下来，我们下调和上调SNHG 9在肝母细胞瘤细胞系中的表达，然后测定细胞增殖（CCK-8）、集落形成和细胞凋亡活性。通过双荧光素酶报告基因活性、RNA免疫沉淀（RIP）、生物素RNA下拉（biotin RNA pull down）和Spemann’s Pearson相关系数分析来确定SNHG 9、Wnt 3a和miR-23 a-5 p之间的相互作用。进行异种移植体内致瘤性试验以阐明SNHG 9肝母细胞瘤在肿瘤发生中的作用。还确定了SNHG 9在肝母细胞瘤中顺铂耐药性中的作用。结果：SNHG 9在肝母细胞瘤组织和细胞系中显著上调。SNHG 9在HUH 6和HepG 2上的过表达导致细胞增殖和克隆形成活性的显著增加，而SNHG 9敲低导致细胞增殖和克隆形成活性的持续抑制。双荧光素酶活性、RNA免疫沉淀和生物素下拉证实了miR-23 a-5 p与SNHG 9的直接相互作用。裸鼠移植瘤实验表明，SNHG 9下调可显著抑制BALB/c小鼠肿瘤的生长。ROC和Kaplan-Meier分析显示SNHG 9在肝母细胞瘤中具有潜在的预后和诊断价值。结论：SNHG 9/miR-23 a-5 p/Wnt 3a轴促进肝母细胞瘤的进展。
Background: Hepatoblastoma is a common hepatic tumor occurring in children between 0-5 years. Accumulating studies have shown lncRNA's potential role in distinct cancer progression and development, including hepatoblastoma. SnoRNA host gene 9 (SNHG9) is associated with the progression of distinct human cancers, but, its specific molecular mechanisms in hepatoblastoma is not unknown. Methods: In this study, we estimated SNHG9 expression in hepatoblastoma tissue and cell lines by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR). Next, we downregulated and upregulated SNHG9 expression in hepatoblastoma cell lines and then determined cell proliferation (CCK-8), colony formation, and cellular apoptosis activity. The dual luciferase reporter activity, RNA immunoprecipitation (RIP), biotin RNA pull down and Spemann's Pearson correlation coefficient assay were performed to establish the interaction between SNHG9, WNt3a and miR- 23a-5p. A xenograft in-vivo tumorgenicity test was performed to elucidate the role of SNHG9 hepatoblastoma in tumorigenesis. SNHG9 role in Cisplatin drug resistance in hepatoblastoma was also determined. Results: SNHG9 was significantly upregulated in hepatoblastoma tissue and cell lines. SNHG9 overexpression on HUH6 & HepG2 resulted in a significant increase in cell proliferation and clonogenic activity while SNHG9 knock down resulted in a sustained inhibition of cell proliferation and clonogenic activity. Dual luciferase activity, RNA immunoprecipitation and biotin pull down confirmed the direct interaction of miR-23a-5p with SNHG9. The xenograft tumorgenicity test showed SNHG9 downregulation significantly inhibited the tumor growth in BALB/c mice. ROC and Kaplan-Meier analysis showed potential prognostic and diagnostic importance of SNHG9 in hepatoblastoma. Conclusion: We concluded that SNHG9/miR-23a-5p/Wnt3a axis promotes the progression hepatoblastoma tumor.