Figure 5.

TNF serves as a major effector in fetal/regenerative/revival cells which exhibit the potential to convert back again into stem-like CRC cells

(A) The protein-protein interaction network of the 2D HC gene set was constructed by STRING and visualized in Cytoscape. Each circle represents a hub node, and the diameter of the circle corresponds with the degree of each node. The largest node, TNF, and the other 11 largest nodes are indicated with gene names.

(B) The quantitative polymerase chain reaction (qPCR) of indicated genes, including typical YAP target genes (F3, CYR61 and CTGF), a fetal marker gene (TACSTD2), and stem cell marker genes (OLFM4, SMOC2 and LGR5) in human colorectal cancer (CRC) organoids cultured under different conditions is shown. The average Ct value of each gene is normalized by the average Ct value of GAPDH, and the normalized value is plotted on the Y axis. The average expression level of seven cultures of 3D MG, 3D MG-TNF, 3D MG-supernatant and 2D HC is presented for each gene. Data are represented as mean ± SD. ∗p < 0.05, paired t-test. 3D MG-TNF; Matrigel CRC organoids were exposed to TNF (50 ng/mL) for 10 days 3D MG-Supernatant; Matrigel CRC organoids were cultured in the 2D HC medium supernatant for 5 days.

(C) The qPCR results for the indicated genes evaluated in (B) are shown. The average expression level of seven cultures of 3D MG, 2D HC and 3D MG from 2D HC is presented for each gene. Data are represented as mean ± SD. ∗p < 0.05, paired t-test. 3D MG from 2D HC; Matrigel CRC organoids which is passaged from 2D HC.