Fig. 5: CAF induced XRCC5 sustains stemness of HCC cells. | Nature Communications

Fig. 5: CAF induced XRCC5 sustains stemness of HCC cells.

From: Multi-omic analysis reveals a CAF-stemness-governed classification in HCC liver transplant recipients beyond the Milan criteria

Fig. 5

A HuH-7 cells and Hep 3B cells with XRCC5 knockdown (shXRCC5-1 and shXRCC5-2) or not were cultured CAF-conditioned medium (CAF+) or not and the expression of XRCC5 was detected using western blotting. Band intensities were semi-quantified using IMAGE LAB software and normalized with β-actin. Values were presented as the means under the bands. B The volume curves of the tumor nodules in groups of HuH-7 cells and HuH-7 cells co-inoculated with CAF in nude mice (n = 7 per group). C Tumor nodules were resected after 20 days and XRCC5 expression were detected by IHC staining. The effect of XRCC5 knockdown or not on stemness of HCC cells were shown by in vitro self-renewal capacity (D), cell migration (E and F), mRNA expression levels of stemness-related genes (G, H) and limiting dilution xenograft formation in NOD/SCID mice (I, shXRCC5 represents XRCC5 knockdown, n = 5 per group), and stem cell frequency was calculated. J In HuH-7 cells and Hep 3B cells with XRCC5 overexpression (OE) or not (vector), XRCC5 was detected using western blotting. The stemness of above cells were shown according to the in vitro self-renewal capacity (K), mRNA expression levels of stemness-related genes (L) and cell migration (M). N IHC staining of XRCC5 in patients with HCC undergoing LT. O Heatmap of differentially expressed genes in the CSC pathway in patients with high or low XRCC5 expression from IHC. P Kaplan‒Meier curves for OS in patients with high or low XRCC5 expression from IHC. The data of cell functional assays and RT-qPCR analysis were presented as mean ± SD of three individual experiments, and the data from animal experiments were presented as mean ± SEM. Two-tailed Student’s t-test was used for comparisons, and P values are shown on the graphs. CAFs cancer-associated fibroblasts, HCC hepatocellular carcinoma; LT liver transplantation, IHC immunohistochemistry, OS overall survival, NC negative control, CI confidence interval. Source data are provided in the Source Data file.

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