扶正抗癌方通过hsa_circ_0048091调控hsa-miR-378g影响PTEN通路抑制非小细胞肺癌增殖的机制研究

Lung cancer ranks the first cause of cancer-related death, and its clinical medication has limitations, so new treatment strategies are urgently needed. Our previous clinical studies have found that Fuzheng kang’ai decoction is effective in the treatment of non-small cell lung cancer (NSCLC), the main type of lung cancer. And our basic studies have found that Fuzheng kang’ai decoction can inhibit the proliferation of NSCLC, but its mechanism is unclear. Our recent preliminary experiments showed that the expression of hsa_circ_0048091 increased after treatment with Fuzheng kang’ai decoction, and silence of hsa_circ_0048091 could promote the proliferation of non-small cell lung cancer cells. At the same time, the expression of miR-378g, which has a binding site of hsa_circ_0048091, was decreased. And the expression of PTEN, which might be regulated by miR-378g, was increased. Therefore, Fuzheng kang’ai decoction can regulate hsa-miR-378g through hsa_circ_0048091 to inhibit the proliferation of non-small cell lung cancer through PTEN pathway. We will use methods including FISH, Pull-down and double luciferase reporter assay, etc. to verify the molecular mechanism of Fuzheng kang’ai decoction in the treatment of non-small cell lung cancer. The completion of this project is expected to provide new biomarkers and therapeutic targets for non-small cell lung cancer, and provide new therapeutic strategies with clear mechanisms.

肺癌位居癌症相关性死因首位，其临床用药存在局限性，迫切需要新的治疗策略。我们前期临床研究发现扶正抗癌方对肺癌的主要类型非小细胞肺癌治疗有效，基础研究发现扶正抗癌方可抑制非小细胞肺癌增殖，但其作用机制不清晰。近期预实验我们发现经扶正抗癌方处理后hsa_circ_0048091表达升高，且沉默它可促进非小细胞肺癌细胞增殖。同时，经扶正抗癌方处理后，与hsa_circ_0048091有结合位点的miR-378g表达下调；且可能被miR-378g调控的PTEN表达上调。因此提出扶正抗癌方可通过hsa_circ_0048091调控hsa-miR-378g影响PTEN通路抑制非小细胞肺癌增殖的科学假说。我们将采用FISH、Pull-down以及双荧光素酶报告基因等方法，来验证扶正抗癌方治疗非小细胞肺癌的分子机制。本项目的完成有望为非小细胞肺癌提供新的分子标记物及治疗新靶点，并提供机理清晰的治疗新策略。

肺癌仍然是位居癌症相关性死因首位的癌种，大部分患者确诊时已处于晚期，失去手术机会。非小细胞肺癌作为肺癌的主要病理类型严重危害国民健康，目前已有的治疗手段包括放化疗、免疫及靶向治疗因严重副作用、药物耐药及应答率低严重制约患者5年生存率的提高，因此迫切需要新的治疗策略。中医药是我国瑰宝，治疗肿瘤历史悠久。前期我们的临床研究证实中药复方扶正抗癌方治疗非小细胞肺癌临床疗效确切，进一步的基础研究发现扶正抗癌方可抑制非小细胞肺癌增殖，但其作用机制不明。本项目中，我们从表观遗传学特别是circRNA角度创新性地探讨了扶正抗癌方抑制非小细胞肺癌增殖的具体分子机制。通过circRNA 测序、miRNA 测序、qRT-PCR、BaseScope、细胞转染、病毒转染、细胞增殖实验、细胞流式、western blot、构建质粒及稳定表达细胞株、荧光素酶报告基因分析、Pull-down assay、FISH、IHC 以及构建肺癌裸鼠皮下成瘤模型等实验方法，从体外细胞、体内动物模型以及临床病人样本三个层面，本项目阐明了扶正抗癌方通过诱导环状RNA hsa_circ_0048091的表达，进而hsa_circ_0048091“海绵”调控 hsa-miR-378g 的表达，hsa-miR-378g 通过影响其靶点ARRDC3的表达，最终明确了扶正抗癌方通过调控hsa_circ_0048091/hsa-miR-378g/ARRDC3通路发挥其抑制非小细胞肺癌增殖的具体分子机制。该项目的完成将为非小细胞肺癌的临床治疗提供新策略及新靶点，为中药复方扶正抗癌方的临床应用提供客观科学依据，具有重要的转化医学价值。