Fig. 10

Role of SP1 in CSC-mediated regulation of ZFAS1. A qRT-PCR analysis of S1  SP1 expression in NREC and lung cancer cells. SP1 expression is much higher in lung cancer cells compared with NREC. Short term CSC exposure does not appear to increase SP1 expression in these cells. B qChIP analysis of SP1 occupancy in the ZFAS1 enhancer in lung cancers and paired normal adjacent lung tissues from smokers and nonsmokers. C qChIP analysis demonstrating that CSC exposure induces recruitment of SP1 to the putative enhancer of ZFAS1 in SAEC and Calu-6 cells. D qChIP analysis demonstrating that knockdown of SP1 decreases endogenous ZFAS1 expression and markedly attenuates CSC-mediated upregulation of ZFAS1 in NREC and lung cancer cells. E, F qRT-PCR analysis demonstrating that mithramycin mediates dose-dependent reduction of ZFAS1 with a concomitant increase in ZNFX1 expression in A549 lung cancer cells in vitro (E) and in vivo (F). Data are mean ± SEM; T-test; n = 3; * p < 0.05; ** p < 0.01