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. 2017 May 30;11(8):981–995. doi: 10.1002/1878-0261.12066

Figure 4.

Figure 4

AURKA‐mediated regulation of c‐MYC and HDM2 requires eIF4E. (A) AGS CDDPR Pool 1, AGS CDDPR Pool 2, AGS CDDPR Clone 1, and AGS CDDPR Clone 2 cells were transiently transfected with control siRNA (siControl) or siRNA specific for eIF4E (sieIF4E) for 48 h. Cell lysates were subjected to western blot analysis of the indicated proteins. Knocking down of eIF4E decreased c‐MYC and HDM2 protein expression. (B) AGS Parental cells were transiently transfected by siControl + AdControl, siControl + AdAURKA, sieIF4E + AdControl, or sieIF4E + AdAURKA for 48 h. Cell lysates were then subjected to western blot analyses of the indicated proteins. The data showed that overexpression of AURKA increased protein levels of p‐eIF4E (S209), c‐MYC, and HDM2. Knockdown of eIF4E suppresses AURKA‐induced upregulation of c‐MYC and HDM2 protein expression. (C) AGS CDDPR Pool 1 and AGS CDDPR Clone 1 cells were transfected with siControl or sieIF4E for 48 h and subjected to CellTiter‐Glo viability assay. Knocking down of eIF4E significantly sensitized cells to CDDP (P < 0.05), as indicated by two‐ to threefold decrease in CDDP IC50. (D) AGS CDDPR Pool 1 and AGS CDDPR Clone 1 cells were transfected with siControl or sic‐MYC for 48 h and subjected to CellTiter‐Glo viability assay. Knocking down of c‐MYC significantly sensitized cells to CDDP (P < 0.05), as indicated by twofold decrease in CDDP IC50.