莲甲散结方通过hsa_circ_0002177/miR-7轴靶向抑制EGFR治疗NSCLC的机制研究

According to Professor Zhou Zhongying's clinical practice, Lian-Jia-San-Jie-Fang (LJSJF) has a significant clinical effect on non-small cell lung cancer. Our previous study demonstrated that LJSJF inhibits tumor by inhibiting EGFR. The pre experiment showed that LJSJF could inhibit the growth of lung cancer cells and regulate EGFR via miR-7. We screened out the significant down – regulated circular RNAs by chip analysis. Bioinformatics analysis shows that miR-7 has a binding site with hsa_circ_0002177, which is one of the down-regulated cicrRNAs. We speculate that LJSJF plays an anti-tumor role by the suppression of EGFR via the hsa_circ_0002177/miR-7 axis. This research plan that NSCLC cells were separately and co transfected by Hsa_circ_0002177 overexpression and shRNA, miR-7 mimics and inhibitors, and LJSJF was given to the cells. Elucidate the mechanism of action from molecular level, cell level, animal model and so on. It provides a theoretical basis to optimize the compatibility of LJSJF, and to find potential active components and molecular targets.

根据周仲瑛教授临床实践总结凝练的莲甲散结方（LJSJF），对NSCLC具有显著的临床疗效。我们既往阐明了LJSJF通过抑制EGFR发挥作用，预实验进一步提示LJSJF体外通过miR-7调控EGFR抑制NSCLC增殖，芯片分析筛选LJSJF处理组小鼠瘤体内显著下调的环状RNA，生物信息分析miR-7与下调环状RNA中的hsa_circ_0002177有结合位点，检测患者NSCLC标本中hsa_circ_0002177的表达显著上调，我们推测LJSJF通过hsa_circ_0002177/miR-7轴抑制EGFR发挥抗肿瘤作用。本研究拟通过分别及共同转染hsa_circ_0002177过表达和shRNA以及miR-7模拟物和抑制物，予LJSJF干预，从分子水平、细胞水平、动物模型予以阐明其之间的相互作用机理，为进一步优化中药配伍，发现潜在有效成分和分子靶点提供理论依据。

根据周仲瑛教授临床实践总结凝练的莲甲散结方（LJSJF），对NSCLC具有显著的临床疗效。我们既往阐明了LJSJF通过抑制EGFR发挥作用，预实验进一步提示LJSJF体外通过miR-7调控EGFR抑制NSCLC增殖，芯片分析筛选LJSJF处理组显著下调的环状RNA，生物信息分析miR-7与下调环状RNA中的hsa_circ_0002177有结合位点，检测患者NSCLC标本中hsa_circ_0002177的表达显著上调。本研究构建了人源性非小细胞肺癌斑马鱼PDX模型，筛选出莲甲散结方对斑马鱼异种移植模型最大安全剂量，无论是肿瘤细胞增殖还是迁移，莲甲散结方均显著的抑制效应。本研究通过分别及共同转染hsa_circ_0002177过表达和shRNA以及miR-7模拟物和抑制物，予LJSJF干预。通过体外模型，阐明了hsa_circ_0002177靶向miR-7促进EGFR表达升高促进NSCLC增殖，侵袭和转移。LJSJF可通过hsa_circ_0002177/miR-7/EGFR信号轴抑制NSCLC进展。通过小鼠体内荷瘤模型证明了LJSJF可抑制NSCLC进展，其可通过调控hsa_circ_0002177/miR-7/EGFR信号轴发挥生物学效应。为进一步优化中药配伍，发现潜在有效成分和分子靶点提供理论依据。斑马鱼PDX模型为临床中医药疗效预测以及精准治疗提供广泛的应用前景。