HUP-T3

ANP32E Downregulation Enhances the Susceptibility of PDAC Cells to Gemcitabine In Vitro

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive and fatal malignancy, although gemcitabine is administered as a single or combined therapeutic agent. Previous studies have demonstrated that ANP32E overexpression promoted PDAC cell proliferation. However, whether it affects treatment outcome and clinical prognosis is still unclear. In the present study, we aimed to determine whether ANP32E is negatively associated with the treatment outcome of gemcitabine.

We downregulated ANP32E expression via specific siRNAs in two PDAC cell lines, HUP-T3 and SU86.86, followed by the detection of gemcitabine-induced cytotoxicity via MTS assays. As shown by the results of the qRT‒PCR (Fig. 1A) and Western blot (Fig. 1B) assays, ANP32E was efficiently knocked down in both cell lines. The MTS assay findings indicated that the downregulation of ANP32E significantly increased gemcitabine-induced cytotoxicity in Hup-T3 and SU86.86 cells (Fig. 1C). In addition, plate cloning experiments revealed that knocking down ANP32E effectively reduced the colony formation ability of PDAC cells and increased their sensitivity to gemcitabine (Fig. 1D).

Owing to the migratory role of ANP32E in PDAC cells, we used a wound healing assay and a transwell migration assay to assess changes in the migration of PDAC cells after the downregulation of ANP32E. The results from the wound healing assay revealed that the migration of pancreatic cancer cells decreased after ANP32E was knocked down. The migration of PDAC cells diminished with increasing concentrations of the drug after the administration of gemcitabine at different concentrations. The antimigration effect was more obvious after the knockdown of ANP32E (Fig. 2A). Similarly, in the Transwell migration assay, downregulation of ANP32E expression in PDAC cells significantly reduced the migratory activity of PDAC cells and increased their sensitivity to gemcitabine (Fig. 2B).