Fig. 7

RT-qPCR analysis of EMT genes. In both 5-FU-treated Caco2 and HT-29 cells, there was a notable shift in the expression of genes related to EMT. In Caco2 cells, TWIST1, ZEB1, and N-cadherin were significantly increased, while SNAIL1, Vimentin, and E-cadherin were decreased following exposure to 5-FU. Importantly, TWIST1 (p-value = 0.0073) and ZEB1 (p-value < 0.0001) were markedly upregulated in 5-FU-exposed Caco2 cells compared to spheroids. Similarly, in HT-29 cells, all EMT-inducing genes were upregulated, while E-cadherin was downregulated. Notably, TWIST1 (p-value < 0.0001) and SNAIL1 (p-value = 0.0049) were significantly elevated in spheroids compared to 5-FU-exposed HT-29 cells. The baseline gene expression in parental cells is represented by a dotted line. Data are presented as mean ± SD from three independent experiments as **** = p-value < 0.0001, ** = p-value < 0.01