Title:Genome-Wide CRISPR-Cas9 Screening and Identification of Potential
Genes Promoting Prostate Cancer Growth and Metastasis
Volume: 23
Issue: 1
关键词:
全基因组，CRISPR-Cas9, DU145，前列腺癌，转移，筛选。
摘要:
Objective: Identification and validation of genes that functionally account for the growth
and metastasis of prostate cancer.
Methods: DU145-KO cell line was constructed by transfecting DU145 cells with lentivirus packaged
with the genome-wide knock-out library. The DU145-KO cells were transplanted into the armpits of
immunocompromised Nu/Nu mice, followed by the tissue collection from the lung at week 3 (early
lung tissue) or week 7 (late lung tissue with micro-metastasis), as well as from primary tumor site at
week 7 (late primary tumor) after inoculation. Lung metastasis was retrieved at various time points
for DNA sequencing analysis to identify enriched sgRNAs, thus candidate genes/miRNAs. Further
bioinformatics analysis and limited functional validation studies were carried out.
Results: DU145-KO cells promoted the formation of transplanted tumors in mice and promoted the
growth and metastasis of primary tumors, compared to the controls (DU145-NC cells). The analysis
of sequence data showed that the abundance of sgRNAs significantly changed in the primary tumor
and micro-metastasis site. Fifteen target genes (C1QTNF9B, FAM229A, hsa-mir-3929, KRT23,
TARS2, CRADD, GRIK4, PLA2G15, LOXL1, SLITRK6, CDC42EP5, SLC2A4, PTGDS, MYL9
and ACOX2 for the enriched sgRNAs) have been selected for experimental validation, which showed
that knock-out of any of these genes led to the enhanced potential of invasion and metastasis of
DU145 cells.
Conclusion: Genome-wide CRISPR-Cas9 knock-out screening technology combined with highthroughput
sequencing analysis identified genes that potentially relate to prostate tumor invasion and
metastasis. Analysis of these genes provided insights into biological pathways relevant to the disease
and disclosed innovative markers for diagnosis or prognosis as well as potential targets for therapy.
