The recent exponential increase in the number of next generation sequencing studies provides a new source of data for the discovery of functional genomics based markers. The RNA-seq and small RNA-seq provide a new source for the discovery of differentially expressed SSRs (simple sequence repeats) as biomarkers in various diseases. In the present study, for the first time, we applied RNA-seq SSR to find new biomarkers for pancreatic cancer (PC) diagnosis. Analysis of RNA-seq data revealed a significant alternation in the frequency of SSR motifs during cancer progression. In particular, RNA-seq SSR showed an increase in the frequencies of GCC/GGC and GCG/CGC motifs in PC samples compared to healthy pancreas. These findings were further confirmed using meta-analysis of EST-SSR data in 11 different cancers. Interestingly, the genes containing GCC/GGC and GCG/CGC motifs in their sequences were involved in many cancer-related biological processes, particularly regulation processes. The small RNA-seq data were also mined for the conserved patterns in SSR frequencies (sRNA-seq SSR) during cancer progression. Based on the results, we suggest the potential use of GCC/GGC and GCG/CGC motifs as biomarkers in PC. Based on the findings of this study, it seems that RNA-seq SSR and sRNA-seq SSR could open a new paradigm in the diagnostic and even therapeutic strategies for PC along the other types of cancers.