Long non-coding RNA AGAP2-AS1, functioning as a competitive endogenous RNA, upregulates ANXA11 expression by sponging miR-16-5p and promotes proliferation and metastasis in hepatocellular carcinoma (Retracted Article)
Long non-coding RNA AGAP2-AS1, functioning as a competitive endogenous RNA, upregulates ANXA11 expression by sponging miR-16-5p and promotes proliferation and metastasis in hepatocellular carcinoma (Retracted Article)
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长非编码RNA AGAP2-AS1作为竞争性内源RNA，通过海绵miR-16-5p上调ANXA11表达并促进肝细胞癌的增殖和转移
DOI：
10.1186/s13046-019-1188-x
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发表时间：
2019-05-14
影响因子：
11.3
通讯作者：
Liu, Qingguang
中科院分区：
文献类型：
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作者：
Liu, Zhikui;Wang, Yufeng;Liu, Qingguang
越来越多的证据强调了长链非编码rna （lncRNAs）在肝细胞癌（HCC）生物学行为中的潜在作用。在此，我们阐明了lncRNA-AGAP2-AS1对HCC生物学行为影响的功能和可能的分子机制。方法采用sedu、Transwell和流式细胞术检测肝癌细胞的增殖、迁移、侵袭和凋亡情况。建立裸鼠皮下肿瘤模型和肺转移小鼠模型，检测肝癌在体内的肿瘤生长和转移情况。荧光素酶报告基因检测证实miR-16-5p与ANXA11的3'UTR直接结合。实时荧光定量PCR检测AGAP2-AS1和miR-16-5p在HCC标本和细胞系中的表达。AGAP2-AS1与miR-16-5p的相关性通过双荧光素酶报告基因实验、RIP实验和生物素下拉实验揭示。结果AGAP2-AS1在HCC组织和细胞系中表达上调，尤其是在转移和复发病例中。体外和体内的功能增益和功能丧失实验表明，AGAP2-AS1促进细胞增殖、迁移、侵袭、EMT进展，抑制HCC细胞凋亡。进一步的研究表明，AGAP2-AS1可以通过在HCC细胞中海绵化miR-16-5p作为竞争内源性RNA （ceRNA）发挥作用。功能、功能获得和功能丧失研究表明，miR-16-5p促进HCC进展，miR-16-5p的改变消除了AGAP2-AS1对HCC细胞的促进作用。此外，ANXA11被确定为HCC细胞中miR-16-5p的直接下游靶点，并介导miR-16-5p和AGAP2-AS1在HCC中的功能作用，导致AKT信号激活。在临床上，AGAP2-AS1和miR-16-5p的表达与HCC患者的不良临床特征和不良预后显著相关。我们发现缺氧是HCC中AGAP2-AS1过表达的原因。缺氧对肝癌细胞转移和EMT的促进作用被AGAP2-AS1敲低逆转。综上所述，本研究首次证实AGAP2-AS1通过AGAP2-AS1/miR-16-5p/ANXA11/AKT轴通路在HCC中起致瘤作用，为HCC患者提供了一种有前景的治疗策略。
BackgroundAccumulating evidence has highlighted the potential role of long non-coding RNAs (lncRNAs) in the biological behaviors of hepatocellular carcinoma (HCC). Here, we elucidated the function and possible molecular mechanisms of the effect of lncRNA-AGAP2-AS1 on the biological behaviors of HCC.MethodsEdU, Transwell and flow cytometry were used to determine proliferation, migration, invasion and apoptosis of HCC cells in vitro. The subcutaneous tumor model and lung metastasis mouse model in nude mice was established to detect tumor growth and metastasis of HCC in vivo. The direct binding of miR-16-5p to 3'UTR of ANXA11 was confirmed by luciferase reporter assay. The expression of AGAP2-AS1 and miR-16-5p in HCC specimens and cell lines were detected by real-time PCR. The correlation among AGAP2-AS1 and miR-16-5p were disclosed by a dual-luciferase reporter assay, RIP assay and biotin pull-down assay.ResultsHere, we demonstrated that AGAP2-AS1 expression was up-regulated in HCC tissues and cell lines, especially in metastatic and recurrent cases. Gain- and loss-of-function experiments indicated that AGAP2-AS1 promoted cell proliferation, migration, invasion, EMT progression and inhibited apoptosis of HCC cells in vitro and in vivo. Further studies demonstrated that AGAP2-AS1 could function as a competing endogenous RNA (ceRNA) by sponging miR-16-5p in HCC cells. Functionally, gain- and loss-of-function studies showed that miR-16-5p promoted HCC progression and alteration of miR-16-5p abolished the promotive effects of AGAP2-AS1 on HCC cells. Moreover, ANXA11 was identified as direct downstream targets of miR-16-5p in HCC cells, and mediated the functional effects of miR-16-5p and AGAP2-AS1 in HCC, resulting in AKT signaling activation. Clinically, AGAP2-AS1 and miR-16-5p expression were markedly correlated with adverse clinical features and poor prognosis of HCC patients. We showed that hypoxia was responsible for the overexpression of AGAP2-AS1 in HCC. And the promoting effects of hypoxia on metastasis and EMT of HCC cells were reversed by AGAP2-AS1 knockdown.ConclusionsTaken together, this research supports the first evidence that AGAP2-AS1 plays an oncogenic role in HCC via AGAP2-AS1/miR-16-5p/ANXA11/AKT axis pathway and represents a promising therapeutic strategy for HCC patients.