Long non-coding RNA CRNDE promotes the proliferation, migration and invasion of hepatocellular carcinoma cells through miR-217/MAPK1 axis.
Long non-coding RNA CRNDE promotes the proliferation, migration and invasion of hepatocellular carcinoma cells through miR-217/MAPK1 axis.
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DOI：
10.1111/jcmm.13856
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发表时间：
2018-12
影响因子：
5.3
通讯作者：
Ma K
中科院分区：
文献类型：
--
作者：
Wang H;Ke J;Guo Q;Barnabo Nampoukime KP;Yang P;Ma K
肝细胞癌是一种侵袭性恶性肿瘤，是全球第二大癌症相关死亡原因。CRNDE和miR-217是在细胞生长、增殖、迁移过程中起关键作用的非编码RNA。丝裂原活化蛋白激酶1(MAPK1)也参与癌细胞过程。因此，本研究旨在探讨CRNDE在肝癌细胞迁移和侵袭中的作用，以及miR-217和MAPK1在这一过程中的作用。基于微阵列的lncRNA图谱研究证实了CRNDE的过表达。定量逆转录聚合酶链式反应证实CRNDE在肝细胞癌中的表达。采用四甲基偶氮唑盐比色法和BrdU染色检测细胞增殖水平。用Transwell法检测细胞的迁移和侵袭能力。创面愈合实验进一步探讨细胞迁移能力。生物信息学预测了MIR-217。通过双荧光素酶报告实验证实CRNDE、miR-217和MAPK1之间的靶向关系。利用生物信息学预测miR-217的下游靶点MAPK1，并通过qRT-PCR和Western印迹进一步证实。采用qRT-PCR、Western印迹、四甲基偶氮唑蓝、BrdU、Transwell实验和伤口愈合实验研究CRNDE、miR-217与MAPK1的相互作用。CRNDE在肝癌组织和肝癌细胞系中表达上调。CRNDE的高表达促进了细胞的增殖、迁移和侵袭，而抑制的CRNDE则相反。MIR-217与CRNDE表达呈负相关，是CRNDE的靶点，在肝细胞癌中表达较低。随着miR-217的高表达，肝癌细胞的增殖、迁移和侵袭受到抑制。MiR-217的可能靶点MAPK1与miR-217呈负相关，与CRNDE呈正相关，在肝细胞癌的形成过程中与CRNDE具有相同的作用。长非编码RNA CRNDE通过miR-217/MAPK1轴促进肝癌细胞的增殖、迁移和侵袭。
Hepatocellular carcinoma (HCC) is an invasive malignant tumour and the second major cause of cancer‐related deaths over the world. CRNDE and miR‐217 are non‐coding RNAs which play critical roles in cell growth, proliferation, migration. Mitogen‐activated protein kinase 1 (MAPK1) also participates in cancer cell process. Hence, this study aimed at investigating the effect of CRNDE on migration and invasion of HCC and figuring out the role of miR‐217 and MAPK1 in this process. The overexpression of CRNDE was demonstrated by a microarray‐based lncRNA profiling study. CRNDE expression in HCC was verified by qRT‐PCR. MTT assay and BrdU staining were applied to detect cell proliferation level. Transwell assay was utilized to examine cell migration and invasiveness abilities. Wound healing assay was performed for further exploration of cell migration capacity. MiR‐217 was predicted by bioinformatics. The dual luciferase reporter assay was performed to corroborate the targeting relationship between CRNDE, miR‐217 and MAPK1. MAPK1, the downstream target of miR‐217, was predicted using bioinformatics and was further confirmed by qRT‐PCR and Western blot. The interaction between CRNDE, miR‐217 and MAPK1 was studied by qRT‐PCR, Western blot, MTT, BrdU, transwell assay and wound healing assay. CRNDE was up‐regulated in HCC tissues and HCC cell lines. The high expression of CRNDE facilitated cell proliferation, migration and invasion, while the inhibited one affected on the contrary. MiR‐217, negatively correlated with CRNDE expression, was the target of CRNDE and was more lowly expressed in HCC. With the high expression of miR‐217, HCC cell proliferation, migration and invasion were suppressed. MAPK1, the possible target of miR‐217, was negatively correlated with miR‐217 but positively correlated with CRNDE and had the same effect in HCC formation process as CRNDE. Long non‐coding RNA CRNDE promotes the proliferation, migration and invasion of HCC cells through miR‐217/MAPK1 axis.
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