In this study, utilizing well established acquired chemoresistant cancer cell lines and their respective parental ones, we provide a series of complementary evidences to show that Smo may promote acquired chemoresistance by activating Gli through Gαi and Gβγ-JNK signaling axis, therefore revealing that GPCR-like signaling elicited from Smo is involved in the canonical Hedgehog-Gli signaling pathway activation and the acquired chemoresistance. The gene discussed is SMO; the disease is cancer.