A transcript analysis was performed using qPCR with primers against a set of 12 genes, including 5 putative CSC markers, LGR5, CD44, ALDH1, CD24, and CD54; 4 stemness regulators, NANOG, OCT4, SOX2 and AICDA; and 3 EMT inducers, PRRX1, TWIST1, and BMI1. The qPCR results showed a significant up-regulation of only LGR5, CD44 and PRRX1 (>2-fold), and not the other genes, in cancer tissues (Fig 1A). This evidence concerns the gene TWIST1 and cancer.