Abstract: Objective To investigate the inhibitory effect of GANT61 on the growth of human lung carcinoma cell lines H1703 and A549 and elucidate its mechanism. Methods H1703 and A549 cells were cultured in vitro. After adding different concentrations of GANT61, the cells growth inhibition rates of H1703 and A549 cell lines were detected by MTS method after 72h. The protein expressions of Gli-1and Gli2 were observed in H1703 and A549 cell lines by Western blot. Transwell invasion assay was performed to evaluate the effect of GANT61 on the invasion ability of H1703 and A549 cell lines. Results MTS method showed that IC50 of GANT61 were 8.6 and 8.2μmol/L in H1703 and A549 cell lines. GANT61 inhibited cell viability of H1703 and A549 cell lines in a dose-dependent manner. GANT61 decreased the expression of Gli1 and Gli2 proteins by Western blot assay. Furthermore, Transwell invasion assay showed that GANT61 suppressed the invasion of H1703 and A549 cell lines significantly. Conclusion GANT61 could effectively inhibit the viability and invasion of H1703 and A549 cell lines, which signified that Gli1 and Gli2 of Hedgehog pathway may play an important role in the carcinogenesis and progression of lung cancer. Gli may become a new antitumor target.