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

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。

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

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.