Gemcitabine is a nucleoside analog that is indicated in the treatment of pancreatic cancer. In order to provide a better use of this drug, the search for immunohistological markers is a hot topic in the field of pancreatic cancer. In particular, the use of nucleoside transporter hENT1 and the intracellular target of gemcitabine RRM1 are current subjects for discussion. We have analyzed the majority of studies of hENT1 and RRM1 on pancreatic cancer, and will discuss the further directions that might be followed in order to integrate these proteins in routine clinical practice. The data that is currently available would benefit from the completion of well-designed randomized trials in order to confirm the clinical value of hENT1 and RRM1 as biomarkers in pancreatic cancer patients.