Increased mRNAs of cancer upregulated gene (CUG)2 were detected in many different tumor tissues using the Affymetrix microarray and the oncogenic capability of the CUG2 gene was further reported. The mechanism by which CUG2 overexpression promotes cancer stem cell (CSC)-like phenotypes, on the other hands, is still unknown. With recent studies showing that pyruvate kinase muscle 2 (PKM2) is overexpressed in clinical tissues from gastric, lung, and cervical cancer patients, we hypothesized that PKM2 plays an important role in CSC-like phenotypes caused by CUG2 overexpression. Herein, compared with the control cells, enhanced PKM2 protein levels and translocation of PKM2 into the nucleus were detected for CUG2-overexpressing lung carcinoma A549 and immortalized bronchial BEAS-2B cells. In the CUG2-overexpressing cells, the expression of c-Myc, CyclinD1, and PKM2 was increased. Furthermore, EGFR and ERK inhibitors as well as the suppression of Yap1 and NEK2 expression reduced PKM2 protein levels. Interestingly, the knockdown of -catenin expression failed to reduce the PKM2 protein levels. Furthermore, the reduction of PKM2 expression with its siRNA hindered CSC-like phenotypes such as faster wound healing, aggressive transwell migration, increased size and number of sphere formation. The introduction of mutant S37A PKM2-green fluorescence protein (GFP) into cells, that were unable to move to the nucleus, in particular, did not confer CSC-like phenotypes, whereas the forced expression of wild-type PKM2 promoted those phenotypes. Overall, CUG2-induced increased expression of nuclear PKM2 contributes to CSC-like phenotypes via upregulation of c-Myc and CyclinD1 as a co-activator.