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

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Ecadherin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。

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.