异甘草素通过lncRNA MIR22HG/miR-24-3p/FASLG轴抑制肺鳞癌转移的机制研究

Mechanism Study of Isoliquiritigenin Inhibiting Lung Squamous Cell Carcinoma Metastasis through lncRNA MIR22HG/miR-24-3p/FASLG Axis

目的 肺鳞癌(Lung squamous cell carcinoma,LUSC)是全世界范围内高频率发生的恶性肿瘤。最近的研究表明,异甘草素(Isoliquiritigenin,ISL)在肿瘤增殖和转移中发挥抗肿瘤活性。因此,研究旨在探讨ISL在抗LUSC转移中的作用机制。方法 利用梯度浓度ISL处理LUSC细胞,探究ISL在LUSC细胞增殖、迁移和侵袭中的抗癌特性。随后,利用生物信息学手段探寻了长链非编码RNA(Long non-coding RNAs, lncRNA)MIR22HG/miR-24-3p/Fas配体基因(Fas ligand Gene,FASLG)之间可能的互作关系。实时荧光定量PCR(Quantitative real-time polymerase chain reaction, qRT-PCR)检测LUSC细胞中lncRNA MIR22HG、miR-24-3p、FASLG的表达,细胞计数盒-8(Cell Counting Kit-8,CCK-8)、克隆形成、划痕愈合和Transwell实验分别检测细胞活力、增殖、迁移和侵袭。蛋白质免疫印迹法(Western blot, WB)检测上皮间充质转换(Epithelial-mesenchymal transition, EMT)和转移相关蛋白的表达。RNA免疫沉淀(RNA Binding Protein Immunoprecipitation, RIP)实验和双萤光素酶实验验证lncRNA MIR22HG与miR-24-3p、miR-24-3p与FASLG的结合关系。结果 ISL可以显著抑制LUSC细胞增殖、迁移和侵袭。生信分析及临床样本分析发现lncRNA MIR22HG在LUSC中低表达,ISL可以促进lncRNA MIR22HG的表达,抑制LUSC细胞的恶性行为。此外,lncRNA MIR22HG可以海绵吸附miR-24-3p进而调节FASLG的表达。miR-24-3p在LUSC中上调表达,FASLG在LUSC中下调表达。过表达FASLG可以逆转miR-24-3p过表达对LUSC增殖、转移以及EMT进程的促进作用。结论 这些结果表明,ISL通过lncRNA MIR22HG/miR-24-3p/FASLG轴抑制肺鳞癌转移,表明ISL有潜力作为治疗LUSC的新型药物。

Objective Lung squamous cell carcinoma(LUSC)is one of the most frequent malignant tumors in the world.Re-cent studies have shown that isoliquiritigenin(ISL)plays an anti-tumor role in tumor proliferation and metastasis.Therefore,this study aims to explore the mechanism of the role of ISL in anti-LUSC metastasis.Method Gradient concentrations of ISL were used to treat LUSC cells to explore the anticancer properties of ISL in the proliferation,migration and invasion of LUSC cells.Subsequently,bioinformatics methods were used to explore the possible interaction between long non-coding RNAs(ln-cRNA)MIR22HG/miR-24-3p/Fas ligand Gene(FASLG).Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expressions of lncRNA MIR22HG,miR-24-3p and FASLG in LUSC cells.Cell Counting Kit-8(CCK-8),clonal formation,scratch healing and Transwell assay were used to detect cell viability,proliferation,migration and in-vasion,respectively.The expression of epithelial-mesenchymal transition(EMT)and transfer-related proteins was detected by Western blot.The binding relationships between lncRNA MIR22HG and miR-24-3p,miR-24-3p and FASLG were verified by RNA Binding Protein Immunoprecipitation(RIP)assay and dual luciferase assay.Result ISL can significantly inhibit LUSC cell proliferation,migration and invasion.Biogenic analysis and clinical sample analysis showed that lncRNA MIR22HG was low expressed in LUSC,and ISL could inhibit the expression of lncRNA MIR22HG and the malignant behavior of LUSC cells.In ad-dition,lncRNA MIR22HG can regulate the expression of FASLG by sponge adsorption of miR-24-3p.The miR-24-3p was up-regulated in LUSC,and FASLG was down-regulated in LUSC.Over-expression of FASLG can reverse the promoting effect of miR-24-3p over-expression on LUSC proliferation,metastasis and EMT process.Conclusion These results suggest that ISL inhibits LUSC metastasis via the lncRNA MIR22HG/miR-24-3p/FASLG axis,suggesting that ISL has potential as a novel treatment for LUSC.