GANT 61 (GANT61) is an inhibitor for Gli1 and Gli2.

GLI1:5 μM

携带SK-N-AS神经母细胞瘤移植瘤的裸鼠口服饲喂50 mg/kg GANT61,在实验第12天明显抑制肿瘤生长,与对照组相比,肿瘤体积减少63％.在注射GLI1阳性22Rv1前列腺癌细胞的裸鼠中,GANT61诱导肿瘤生长退化,直到观察不到明显的肿瘤.

GANT61以GLI依赖性方式有效抑制体外肿瘤细胞增殖。30 μM GANT61导致急性髓系白血病细胞生长停滞和凋亡。GANT61对人结肠癌细胞系具有强大的细胞毒性,且消除集落生成。GANT61作用于慢性淋巴细胞性白血病细胞,而非正常的B淋巴细胞,诱导细胞凋亡。GANT61在人结肠癌细胞系的早期S期中诱导DNA复制的抑制,导致ATM-Chk2轴的DNA损伤信号传导和诱导细胞死亡。GANT61抑制GLI1的DNA结合能力。GANT61抑制hedgehog信号,IC50为5 μM,比作用于其他通路选择性高（如TNF信号/NFκB激活,糖皮质激素受体基因转录,及Ras-Raf-Mek-Mapk级联）。

Dual Luciferase Assay: HEK293 cells are transfected with GLI1 expression plasmid, together with the reporter plasmids 12×GliBSLuc and R-Luc on 10-cm plates (day 0). Twenty-four hours later, cells are seeded in white 96-well plates with clear bottom at a density of 15,000 cells per well. Cells are allowed to attach, and compounds are added at a final concentration of 10 μM in DMSO (0.5% final DMSO concentration) (day 1.5). Cells are grown for another 24 h, subsequently lysed, and then analyzed by using the Dual Luciferase kit.

BrdU Incorporation Assay. Subconfluent cells are grown in reduced FBS (2.5%) for 48 h in the presence of 5 μM test compound (or DMSO) on white 96-well plates with clear bottom. Subsequently, cells are labeled for 2 h with BrdU, fixed, and analyzed.(Only for Reference)