Figure 3.

Knockdown of lncRNA XIST inhibits EMT and stem cell formation in CRC cells. (a) HCT116 cells underwent morphological change from a spindle shape to a rounded or cobblestone-like shape upon knockdown of lncRNA XIST. (b) Knockdown of lncRNA XIST significantly increased the expression of E-cadherin, whereas it decreased the expression of N-cadherin in SW620 cells as demonstrated by immunofluorescence. (c) Real-time PCR analysis showed that knockdown of lncRNA XIST significantly reduced the expression of mesenchymal markers (N-cadherin, Vimetin, Snail, Slug), whereas it increased the expression of epithelial markers (E-cadherin, α-catenin, β-catenin) (*P<0.05) (n=3 independent experiments). (d) Real-time PCR analysis showed knockdown of lncRNA XIST could markedly decrease the expression of many stemness-associated genes (Nanog, Oct-4 and SOX2) and surface antigens associated with cancer stem cells (CD24, CD44, CD133, CD155 and CD166) (*P<0.05) (n=3 independent experiments). (e) Knockdown of lncRNA XIST significantly reduced the sphere formation in HCT116 and SW620 cells (*P<0.05). Error bars represent the mean±S.D. values