Wnt7a is methylated in NSCLC cell lines and tissues.

<p>A) B2B and HBEC cells were treated with 10 uM NNK for 2 hours and Wnt7a expression was measured by QPCR compared to an untreated control. Error bars are the SEM of triplicate experiments. Percent methylation in B2B and HBEC cells was analyzed using the Methyl Profiler system after 2 hr 10 uM NNK treatment compared to untreated cells. B) Mean percent methylation of the Wnt7a promoter was measured by pyrosequencing in NSCLC tissue and compared to matched normal adjacent lung tissue by paired t-test. Mean percent methylation of B2B, H157, and H1299 cell lines was measured by pyrosequencing. C) A Wnt7a promoter luciferase was modified by SssI, HpaII, and HhaI methylating enzymes and transiently transfected into B2B and HBEC cell lines. Fold-change in luciferase activity was measured compared to unmodified Wnt7a promoter luciferase activity. Error bars are the SE of triplicate experiments.