Department of Thoracic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China.
Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China.
PLoS Genet. 2020 Nov 2;16(11):e1009168. doi: 10.1371/journal.pgen.1009168. eCollection 2020 Nov.
MicroRNAs (miRNAs) play important roles in the development of various cancers including lung cancer which is one of the devastating diseases worldwide. How miRNAs function in de novo lung tumorigenesis remains largely unknown. We here developed a CRISPR/Cas9-mediated dual guide RNA (dgRNA) system to knockout miRNAs in genetically engineered mouse model (GEMM). Through bioinformatic analyses of human lung cancer miRNA database, we identified 16 downregulated miRNAs associated with malignant progression and performed individual knockout with dgRNA system in KrasG12D/Trp53L/L (KP) mouse model. Using this in vivo knockout screening, we identified miR-30b and miR-146a, which has been previously reported as tumor suppressors and miR-190b, a new tumor-suppressive miRNA in lung cancer development. Over-expression of miR-190b in KP model as well as human lung cancer cell lines significantly suppressed malignant progression. We further found that miR-190b targeted the Hus1 gene and knockout of Hus1 in KP model dramatically suppressed lung tumorigenesis. Collectively, our study developed an in vivo miRNA knockout platform for functionally screening in GEMM and identified miR-190b as a new tumor suppressor in lung cancer.
微小 RNA（miRNAs）在各种癌症的发展中发挥着重要作用，包括肺癌，这是全球破坏性疾病之一。miRNAs 在新发生的肺肿瘤发生中的作用在很大程度上仍然未知。我们在这里开发了一种 CRISPR/Cas9 介导的双重向导 RNA（dgRNA）系统，以在基因工程小鼠模型（GEMM）中敲除 miRNAs。通过对人类肺癌 miRNA 数据库的生物信息学分析，我们鉴定了 16 个与恶性进展相关的下调 miRNAs，并使用 dgRNA 系统在 KrasG12D/Trp53L/L（KP）小鼠模型中进行了单独敲除。通过这种体内敲除筛选，我们鉴定了 miR-30b 和 miR-146a，它们先前被报道为肿瘤抑制因子，以及 miR-190b，这是一种在肺癌发展中具有新的肿瘤抑制作用的 miRNA。在 KP 模型以及人类肺癌细胞系中过表达 miR-190b 显著抑制恶性进展。我们进一步发现 miR-190b 靶向 Hus1 基因，并且在 KP 模型中敲除 Hus1 显著抑制肺肿瘤发生。总之，我们的研究开发了一种用于 GEMM 中功能筛选的体内 miRNA 敲除平台，并鉴定了 miR-190b 作为肺癌中的一种新的肿瘤抑制因子。
