Long non-coding RNA HCP5 promotes proliferation and metastasis of clear cell renal cell carcinoma via targeting miR-1 40-5p/IGF1R pathway
Long non-coding RNA HCP5 promotes proliferation and metastasis of clear cell renal cell carcinoma via targeting miR-1 40-5p/IGF1R pathway
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DOI：
10.26355/eurrev_202003_20661
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发表时间：
2020-03-01
影响因子：
3.3
通讯作者：
Lu, C.
中科院分区：
文献类型：
--
作者：
Zhang, Y-J;Lu, C.
目的：材料与方法：采用定量真实的时间-聚合酶链反应（qRTPCR）方法检测HCC组织和细胞中HCP 5和miR-140- 5 p的表达水平。采用Kaplan-Meier生存分析法分析HCP 5对患者预后的影响。CCK-8法检测细胞增殖、凋亡，流式细胞仪检测细胞周期和侵袭能力。通过生物信息学位点预测miR-140- 5 p与HCP 5或IGF 1 R的结合位点，并通过双荧光素酶报告基因分析或RIP-ago 2分析证实miR-140- 5 p与HCP 5或IGF 1 R的直接相互作用。Western blot检测目的基因的表达。建立异种移植模型，验证HCP 5在体内的功能。结果：ccRCC组织和细胞中HCP 5的表达明显上调。此外，与HCP 5低表达的患者相比，HCP 5高表达水平的患者具有不利的总生存期（OS）和无进展生存期（PFS）。此外，HCP 5敲低导致ccRCC细胞增殖、集落形成、迁移和侵袭的抑制;促进体外G 0-G1期细胞周期停滞和细胞凋亡;以及体内肿瘤生长的抑制。在机械上，miR-140- 5 p被证明是HCP 5的抑制性靶标。此外，胰岛素样生长因子-1受体（IGF 1 R）被鉴定为ccRCC细胞中miR-140- 5 p的直接靶点。最后，我们发现HCP 5沉默对功能行为的抑制作用被miR-140- 5 p沉默所中和。结论：HCP 5作为一种竞争性内源性RNA，通过海绵状作用于ccRCC中的miR-140- 5 p来调节IGF 1 R的表达。因此，HCP 5/miR-140- 5 p/IGF 1 R通路可能是ccRCC中有希望的治疗靶点。
OBJECTIVE: The expression pattern, biological function and action mechanism of long noncoding RNA HCP5 in clear cell renal cell carcinoma (ccRCC) remain elusive.MATERIALS AND METHODS: The quantitative Real Time-Polymerase Chain Reaction (qRTPCR) was used to measure the abundance of HCP5 and miR-140-5p in HCC tissues and cells. Kaplan-Meier survival analysis was used to analyze the prognostic role of HCP5 for the patients. Cell proliferation, apoptosis, as well as cell cycle and metastasis were detected by CCK-8, flow cytometry, transwell migration and invasion assays, respectively. The binding sites between miR-140-5p and HCP5 or IGF1R were predicted by bioinformatic sites, and the direct interaction was confirmed by Dual-Luciferase reporter assay or RIP-ago2 assay. Western blot assay was used to detect the expression of target gene. Xenograft model was established to validate the function of HCP5 in vivo.RESULTS: The expression of HCP5 was significantly upregulated in ccRCC tissues and cells. Moreover, patients with high HCP5 expression level had unfavorable overall survival (OS) and progression-free survival (PFS) compared to those with low HCP5 expression. Additionally, HCP5 knockdown led to the prohibition of ccRCC cell proliferation, colony formation, migration, and invasion; the promotion of cell cycle arrest at G0-G1 and apoptosis in vitro; and the inhibition of tumor growth in vivo. Mechanically, miR-140-5p was certified as an inhibitory target of HCP5. Furthermore, insulin-like growth factor-1 receptor (IGF1R) was identified as a direct target of miR-140-5p in ccRCC cells. Finally, we found that the inhibitory effects of the HCP5 silencing on functional behaviours were neutralized by miR-140-5p silencing.CONCLUSIONS: HCP5 serves as a competing endogenous RNA that regulated IGF1R expression by sponging miR-140-5p in ccRCC. Hence, the HCP5/miR-140-5p/IGF1R pathway might be a promising therapeutic target in ccRCC.