Schlafen 3 Mediates the Differentiating Effects of Cdx2 in Rat IEC-Cdx2L1 Enterocytes

Abstract

Aim Mature, differentiated enterocytes are essential for normal gut function and critical to recovery from pathological conditions. Little is known about the factors that regulate intestinal epithelial cell differentiation in the adult intestine. The transcription factor, Cdx2, involved in enterocytic differentiation, remains expressed in the adult. Since we have implicated Slfn3 in differentiation in vivo and in vitro, we examined whether it also mediated differentiation in the IEC-Cdx2-L1 cell model of differentiation. Materials and Methods IEC-Cdx2-L1 cells, permanently transfected with Cdx2 under the control of isopropyl-β-D-thiogalactoside (IPTG), were stimulated to differentiate by 16-day exposure to IPTG. Transcript levels of Cdx2, Slfn 3 and villin were determined by quantitative reverse transcriptase-polymerase chain reaction of mRNA isolated from IPTG-treated and control cells. Slfn3 expression was lowered with specific siRNA to investigate the role of Slfn3 in Cdx2-driven villin expression in IPTG-differentiated cells. Results Slfn3 and villin expression was significantly greater in IPTG-treated cells. Slfn3 siRNA lowered Slfn3 expression and abolished the IPTG-induced rise in villin expression (p<0.05 by ANOVA); Cdx2 expression was unaffected by Slfn3 siRNA. Discussion The data indicate that the presence of Slfn3 is required for Cdx2 to induce villin expression and thus, differentiation. However, Slfn3 must also promote differentiation independently of Cdx2 since IEC-6 cells that do not normally express Cdx2 can be differentiated by a variety of Slfn3-dependent mechanisms.

Schlafen 12 mediates the effects of butyrate and repetitive mechanical deformation on intestinal epithelial differentiation in human Caco-2 intestinal epithelial cells

“…Indeed, Schlafen 3 is required for the induction of villin expression by the differentiating transcription factor Cdx2 in rat IEC-6-Cdx2-L1 cells. [50]. Thus, there may be good reason to link Schlafen 3 to rodent enterocyte differentiation.…”

Schlafen 3 knockout mice display gender-specific differences in weight gain, food efficiency, and expression of markers of intestinal epithelial differentiation, metabolism, and immune cell function

“…However, there were no significant differences between male Slfn3KO and WT mice and no significant differences in Notch1 expression for either sex between Slfn3KO and WT mice (Fig 8A and 8B). Additionally, since we have shown that Slfn3 can affect the expression of Cdx2 in IEC-6 cells and Boyd et al listed Notch2 as a target gene of Cdx2 [1, 31] we explored whether the expression of Cdx2 was affected in Slfn3KO mice. In agreement with the Notch2 expression, Cdx2 was also increased in Slfn3KO female mice in comparison to WT females, while there was no significant change between the male genotypes (Fig 8C).…”

Schlafens: Emerging Proteins in Cancer Cell Biology

“…Slfn3 is expressed preferentially in CD4 + CD25 + Treg cells and downregulates during T cell activation by TGF-β [ 42 ]. Outside the immune system, Slfn3 also regulates enterocytic differentiation [ 28 , 43 , 44 ] and colonic mucosal growth during aging [ 45 ]. Slfn3 knockout mice show gender-specific differences in weight gain, intestinal epithelial differentiation, and impaired enterocyte differentiation after bowel resection [ 46 ].…”