Panoply™ Human DCK Over-expressing Stable Cell Line

Reportedly, reduced expression levels of deoxycytidine kinase (dCK) serve as an indicator of gemcitabine efficacy in pancreatic cancer, a phenomenon attributed to the kinase's influence on gemcitabine metabolism. The transcription factor NF-E2 p45-related factor 2 (NRF2, also known as Nfe2l2)-a master regulator of redox homeostasis-has been reported to strictly govern the expression of numerous reactive oxygen species (ROS) detoxification genes and to play a role in mediating drug resistance. Here, researchers evaluated the impact of dCK on NRF2 transcriptional activity by overexpressing dCK in pancreatic cancer cells. The results demonstrated that dCK exerts a negative regulatory effect on NRF2 transcriptional activity, thereby leading to reduced expression levels of ARE-driven antioxidant genes. Furthermore, dCK was found to negatively regulate intracellular redox homeostasis and ROS generation. In both pancreatic cancer cell lines and patient samples, the researchers observed a negative correlation between the expression levels of dCK and NRF2. In vitro studies using cell lines further suggested that dCK exerts a negative regulatory effect on cellular proliferation and metastasis.

Here, CCK-8 proliferation assays demonstrated that the proliferation of dCK-overexpressing PANC-1 and MIA PaCa-2 cells was inhibited (Figure 1A). Subsequently, the researchers performed colony formation assays, the results of which indicated that the colony-forming ability of dCK-overexpressing PANC-1 and MIA PaCa-2 cells was suppressed (Figure 1B, C). Next, the researchers evaluated the impact of dCK expression on the invasiveness of PANC-1 and MIA PaCa-2 cells, observing that dCK overexpression inhibited the invasive capacity of these cells (Figure 1D, E). Furthermore, apoptosis was promoted in dCK-overexpressing PANC-1 and MIA PaCa-2 cells, suggesting that dCK plays a negative regulatory role in pancreatic cancer cell proliferation (Figure 1F). Finally, the researchers analyzed potential signaling pathways involved in mediating drug resistance and anti-apoptotic effects. These results confirmed that dCK overexpression inhibited the activation of ERK1/2. Additionally, the protein levels of Mcl1-a recognized anti-apoptotic factor known to be involved in mediating resistance to chemotherapy and radiotherapy-were concomitantly reduced (Figure 1G).

Figure 1. dCK suppressed pancreatic cancer cell proliferation. (Hu Q, et al., 2018)