In vivo miRNA knockout screening identifies miR-190b as a novel tumor suppressor
Autoři:
Hui Hong aff001; Shun Yao aff003; Yuanyuan Zhang aff005; Yi Ye aff003; Cheng Li aff007; Liang Hu aff001; Yihua Sun aff001; Hsin-Yi Huang aff003; Hongbin Ji aff003; Liang Hu aff003
Působiště autorů:
Department of Thoracic Surgery, Fudan University Shanghai Cancer Center, Shanghai, China
aff001; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, China
aff002; State Key Laboratory of Cell Biology, Shanghai Institute of Biochemistry and Cell Biology, Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences; Shanghai, China
aff003; University of Chinese Academy of Sciences, Beijing, China
aff004; BIOPIC and School of Life Sciences, Peking University, Beijing, China
aff005; School of Life Science and Technology, Shanghai Tech University, Shanghai, China
aff006; Peking-Tsinghua Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, School of Life Sciences, Peking University, Beijing, China
aff007; Center for Statistical Science, Center for Bioinformatics, Peking University, Beijing, China
aff008
Vyšlo v časopise:
In vivo miRNA knockout screening identifies miR-190b as a novel tumor suppressor. PLoS Genet 16(11): e32767. doi:10.1371/journal.pgen.1009168
Kategorie:
Research Article
doi:
https://doi.org/10.1371/journal.pgen.1009168
Souhrn
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.
Klíčová slova:
Breast cancer – Carcinogenesis – Genetically modified animals – Guide RNA – Lung and intrathoracic tumors – MicroRNAs – Mouse models – Secondary lung tumors
Zdroje
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