长链非编码RNA FOXN3-AS2在肝癌中的表达及其对肝癌细胞增殖和侵袭的影响

doi:10.12092/j.issn.1009-2501.2018.11.008

中国临床药理学与治疗学 ›› 2018, Vol. 23 ›› Issue (11): 1246-1251.doi: 10.12092/j.issn.1009-2501.2018.11.008

长链非编码RNA FOXN3-AS2在肝癌中的表达及其对肝癌细胞增殖和侵袭的影响

蔡民，许浏，沈兰，张杰

嘉兴市第一医院普外科，嘉兴 314001，浙江

收稿日期:2018-07-09 修回日期:2018-07-29 出版日期:2018-11-26 发布日期:2018-11-22
通讯作者: 张杰，男，本科，副主任医师，主要从事lncRNA在肝癌中的作用机制研究。 Tel: 13736403965 E-mail: alenlily1@163.com
作者简介:蔡民，男，硕士，副主任医师，主要从事lncRNA在肝癌中的作用机制研究。
基金资助:
嘉兴市科技计划项目（2015AY23013）

Expression of long-chain non-coding RNA FOXN3-AS2 in hepatocellular carcinoma and its effect on proliferation and invasion of hepatoma cells

CAI Min, XU Liu, SHEN Lan, ZHANG Jie

Department of General Surgery, the First Hospital of Jiaxing, Jiaxing 314001, Zhejiang, China

Received:2018-07-09 Revised:2018-07-29 Online:2018-11-26 Published:2018-11-22

摘要/Abstract

摘要：

目的: 观察FOXN3-AS2基因在肝癌组织和细胞中的表达，探讨其对肝癌细胞增殖和侵袭的影响。方法: 实时荧光定量PCR（qRT-PCR）法检测肝癌组织和细胞株中FOXN3-AS2的表达水平。在表达水平最低的肝癌细胞株转染携带FOXN3-AS2的质粒以升高FOXN3-AS2的表达，细胞计数试剂盒(cell counting kit-8，CCK-8)和Transwell侵袭实验检测FOXN3-AS2高表达对肝癌细胞增殖活性和侵袭能力的影响。生物信息学预测FOXN3-AS2互补结合的miRNA及相关基因，qRT-PCR检测miRNA和相关基因mRNA的表达水平，Western blot检测相关蛋白的表达水平。结果: FOXN3-AS2在肝癌组织的表达水平显著低于癌旁组织（P<0.01），在肝癌细胞株的表达水平显著低于人正常肝细胞（P<0.05），在HepG2细胞的表达水平最低（P<0.01）。FOXN3-AS2高表达可显著抑制肝癌细胞的增殖活性（P<0.05）和侵袭能力（P<0.05）。FOXN3-AS2可互补结合miR-34a-5p，miR-34a-5p可互补结合Kruppel样因子4（KLF4）基因。FOXN3-AS2高表达可降低miR-34a-5p的表达水平（P<0.01），KLF4 mRNA表达水平升高（P<0.01），KLF4、β-catenin和E-cadherin蛋白表达升高，CDK4和Cyclin D1蛋白表达降低。结论: FOXN3-AS2在肝癌组织和细胞中表达降低，FOXN3-AS2高表达可抑制肝癌HepG2细胞的增殖和侵袭，其机制可能是调控miR-34a-5p及KLF4基因的表达。

关键词: 肝细胞癌, 长链非编码RNA, 细胞增殖, 细胞侵袭

Abstract:

AIM: To observe the expression levels of FOXN3-AS2 in hepatocellular tissues and cells, and its effect on proliferation and invasion of hepatoma cells. METHODS: The expression levels of FOXN3-AS2 in 12 hepatocellular carcinoma tissues and 5 hepatocellular carcinoma cell lines were detected by real-time quantitative PCR (qRT-PCR). Transfection of plasmid in the hepatocellular carcinoma cell line with the lowest expression level of FOXN3-AS2 was used to increase the expression of FOXN3-AS2. The cell counting kit (CCK-8) and Transwell invasion assay were used to detect the effect of overexpressed FOXN3-AS2 on proliferation and invasion of hepatoma cells. Bioinformatics was used to predict the miRNA that FOXN3-AS2 could complement and related genes, qRT-PCR detected the expression levels of miRNA and related gene mRNA, and Western blot was used to detect the expression level of related proteins. RESULTS: The expression level of FOXN3-AS2 in hepatocellular carcinoma tissues was significantly lower than that in adjacent tissues (P<0.01). The expression level of FOXN3-AS2 in hepatocellular carcinoma cell lines was significantly lower than that in human normal liver cells (P<0.05). FOXN3-AS2 has the lowest expression level in HepG2 cells (P<0.01). High expression of FOXN3-AS2 significantly inhibited the proliferation activity (P<0.05) and invasive ability (P<0.05). FOXN3-AS2 could complement the miR-34a-5p, and miR-34a-5p can complement the KLF4 gene. FOXN3-AS2 can complementarily pair with miR-34a-5p, and miR-34a-5p can complementarily pair with KLF4. High expression of FOXN3-AS2 decreased the expression of miR-34a-5p (P<0.01) and increased the expression of KLF4 mRNA (P<0.01). The expression of KLF4, β-catenin and E-cadherin proteins were increased, while the expression of CDK4 and Cyclin D1 proteins were decreased. CONCLUSION: The expression of FOXN3-AS2 is down-regulated in HCC tissues and cells. The high expression of FOXN3-AS2 can inhibit the proliferation and invasion of HepG2 cells. The mechanism may be related with regulating the expression of miR-34a-5p and KLF4 genes.

Key words: hepatocellular carcinoma, long non-coding RNA, cell proliferation, cell invasion

中图分类号:

蔡民，许浏，沈兰，张杰. 长链非编码RNA FOXN3-AS2在肝癌中的表达及其对肝癌细胞增殖和侵袭的影响[J]. 中国临床药理学与治疗学, 2018, 23(11): 1246-1251.

CAI Min, XU Liu, SHEN Lan, ZHANG Jie. Expression of long-chain non-coding RNA FOXN3-AS2 in hepatocellular carcinoma and its effect on proliferation and invasion of hepatoma cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2018, 23(11): 1246-1251.

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长链非编码RNA FOXN3-AS2在肝癌中的表达及其对肝癌细胞增殖和侵袭的影响

Expression of long-chain non-coding RNA FOXN3-AS2 in hepatocellular carcinoma and its effect on proliferation and invasion of hepatoma cells

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