Fig. 1: In vivo genome-wide CRISPR-Cas9 screening identifies HLF as a lung metastasis suppressor in ccRCC.
From: SWI/SNF ATPase silenced HLF potentiates lung metastasis in solid cancers
a Schematic outline of whole-genome CRISPR screen to identify essential genes regulating tumor metastasis in xenograft mouse models for kidney cancer. Created in BioRender. Zhou, J. (2025) https://BioRender.com/x5scj3g. b Venn diagram showing the number of overlapping genes essential for UMRC2 and A498 cells metastasizing to the lung, and shared genes were ranked based on their essentiality scores for lung metastasis. The top ten genes with highest essentiality scores for both cell lines were denoted. c, d Quantification of wound healing assay (n = 3 independent cell cultures) (c) and transwell invasion assay (n = 6 independent cell cultures) (d) in 786-O cells transduced with pLX304-empty vector (Ctrl_EV) or target genes cloned into pLX304 backbone. e Schematic showing primary tumor (PT) in kidney and corresponding lung metastatic tumors (Lung mets), representative images of HLF RNAScope assay and HLF IHC assay in patient renal PTs and Lung mets, and corresponding H-score. N = 6 patient samples (n = 3 paired / n = 3 unpaired ccRCC tumors), scale bar, 20 µm for RNAScope (10 µm for zoomed-in views), 50 µm for IHC. Created in BioRender. Zhou, J. (2025) https://BioRender.com/x5scj3g. f Schematic of tumor thrombus (TT) from kidney primary tumor (PT) (left) and HLF mRNA level in TTNM (tumor thrombus, patients with no metastasis) compared with TTM (patients with metastasis), n = 68 in TTNM group, n = 72 in TTM group. Created in BioRender. Zhou, J. (2025) https://BioRender.com/x5scj3g. g Overall survival of HLF from the cancer genome atlas kidney renal clear cell carcinoma (TCGA-KIRC) dataset. Sample sizes are indicated in the figure. h The mRNA levels of HLF in primary versus metastatic tumors across the indicated cancers were analyzed using the TNMplot web tool, sample sizes are indicated in the figure. Data are mean ± s.e.m. (c, e, f), box plots show the median and interquartile range, and whiskers show the data range (d, h), the error band in the Kaplan-Meier survival plot represents the confidence interval of the estimated survival probability at each time point (g). One-way ANOVA followed by a post hoc Dunnett-t-test (d), two-sided Mann-Whitney U test (e), unpaired two-tailed t-test (f) or Dunn’s test following Kruskal–Wallis test (h). Source data are provided as a Source Data file.
