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Mammalian Expression> | … For the 5′TOP activity assay, cells were co-transfected with a monocistronic reporter containing 5′TOP motif of Rps3 or Rpl36 genes (GenScript (Piscataway, NJ, USA) and pRL construct constitutively expressing the Renilla luciferase gene as an internal control for … | Get A Quote |
We previously showed that N6L, a pseudopeptide that targets nucleolin, impairs pancreatic ductal adenocarcinoma (PDAC) growth and normalizes tumor vessels in animal models. In this study, we analyzed the translatome of PDAC cells treated with N6L to identify the pathways that were either repressed or activated. We observed a strong decrease in global protein synthesis. However, about 6% of the mRNAs were enriched in the polysomes. We identified a 5'TOP motif in many of these mRNAs and demonstrated that a chimeric RNA bearing a 5'TOP motif was up-regulated by N6L. We demonstrated that N6L activates the mTOR pathway, which is required for the translation of these mRNAs. An inhibitory synergistic effect in PDAC ce... More