长链非编码RNA NKILA通过靶向miR-485-5p促进肝癌细胞的上皮-间质转化
LncRNA NKILA Promotes Epithelial-Mesenchymal Transition of Liver Cancer Cells by Targeting miR-485-5p.
作者信息
Wang Yuxu, Li Chao, Shi Yuyi, Kuai Jing
机构信息
Department of Hepatobiliary Surgery, Weifang People's Hospital, Weifang 261000, Shandong Province, China.
出版信息
J Oncol. 2021 Sep 3;2021:1281031. doi: 10.1155/2021/1281031. eCollection 2021.
OBJECTIVE
Liver cancer (LC), one of the familiar malignancies, has a very high morbidity all over the world. The onset of the disease is hidden, and the patients usually do not express any special symptoms. Most of them will have been developed to the middle and later stage when they are diagnosed. This is one of the main reasons why the prognosis of LC is extremely pessimistic all the year round. Recently, researchers have focused mainly on molecular studies, among which LncRNA is a hot spot. This research aims to explore the biological behaviors of LncRNA NKILA and miR-485-5p in LC cells and verify the relationship between them, thereby providing a new theoretical basis for future prevention and treatment.
METHODS
Ninety-four early LC patients admitted to our hospital from January 2015 to January 2017 were regarded as the research objects. In addition, human LC cells SMMC-7721, HepG2, and normal liver cells HL-7702 were purchased. The LncRNA NKILA and miR-485-5p level in cancer and adjacent tissues, LC, and normal liver cells of patients was tested by PCR. Patients were followed up for 3 years. Then, LncRNA NKILA and miR-485-5p's effects on prognosis and cell biological behavior were analyzed. At last, the relationship between LncRNA NKILA and miR-485-5p was assessed by a dual-luciferase reporter assay.
RESULTS
The LncRNA NKILA expression was high in LC tissues and cells ( < 0.050), while miR-485-5p was low compared with the normal adjacent tissues ( < 0.050). Prognostic follow-up manifested that high LncRNA NKILA or low miR-485-5p could predict the poor prognosis and high mortality risk of the patients ( < 0.050). LC cells with downregulated LncRNA NKILA documented inhibited proliferation, invasion, and EMT, while the apoptosis level of the cells increased ( < 0.050). The proliferation, invasion, and EMT were inhibited by miR-485-5p increase, while the apoptosis of the cells decreased after upregulating miR-485-5p ( < 0.050). Online websites predicted that LncRNA NKILA had a binding site with miR-485-5p, and dual-luciferase reporter assay confirmed that LncRNA NKILA could directly target with miR-485-5p ( < 0.050). The miR-485-5p in LC cells increased after LncRNA NKILA was silenced ( < 0.050). The rescue experiment documented that LncRNA NKILA inhibition on LC cells was reversed by inhibiting miR-485-5p ( < 0.050).
CONCLUSION
The LncRNA NKILA with high expression advances LC cell proliferation, invasion, and EMT by targeting miR-485-5p.
目的
肝癌（LC）是常见的恶性肿瘤之一，在全球发病率极高。该病起病隐匿，患者通常无特殊症状表现。多数患者确诊时已发展至中晚期。这是LC预后常年极为悲观的主要原因之一。近年来，研究人员主要聚焦于分子研究，其中长链非编码RNA（LncRNA）是一个热点。本研究旨在探讨LncRNA NKILA和miR - 485 - 5p在LC细胞中的生物学行为，并验证它们之间的关系，从而为未来的防治提供新的理论依据。
方法
选取2015年1月至2017年1月我院收治的94例早期LC患者作为研究对象。此外，购买人LC细胞SMMC - 7721、HepG2以及正常肝细胞HL - 7702。采用聚合酶链反应（PCR）检测患者癌组织及癌旁组织、LC细胞及正常肝细胞中LncRNA NKILA和miR - 485 - 5p的水平。对患者进行3年随访。然后，分析LncRNA NKILA和miR - 485 - 5p对预后及细胞生物学行为的影响。最后，通过双荧光素酶报告基因检测评估LncRNA NKILA与miR - 485 - 5p之间的关系。
结果
LC组织和细胞中LncRNA NKILA表达较高（<0.050），而与正常癌旁组织相比，miR - 485 - 5p表达较低（<0.050）。预后随访表明，LncRNA NKILA高表达或miR - 485 - 5p低表达可预测患者预后不良及高死亡风险（<0.050）。LncRNA NKILA表达下调的LC细胞增殖、侵袭及上皮 - 间质转化（EMT）受到抑制，而细胞凋亡水平升高（<0.050）。miR - 485 - 5p表达增加可抑制增殖、侵袭及EMT，上调miR - 485 - 5p后细胞凋亡减少（<0.050）。在线网站预测LncRNA NKILA与miR - 485 - 5p有结合位点，双荧光素酶报告基因检测证实LncRNA NKILA可直接靶向miR - 485 - 5p（<0.050）。LncRNA NKILA沉默后LC细胞中miR - 485 - 5p增加（<0.050）。挽救实验表明，抑制miR - 485 - 5p可逆转LncRNA NKILA对LC细胞的抑制作用（<0.050）。
结论
高表达的LncRNA NKILA通过靶向miR - 485 - 5p促进LC细胞增殖、侵袭及EMT。
Zotero 插件