The nuclear receptor-mediated genomic pathways of the animal steroid hormones are well known. However, the cell membrane receptor-mediated nongenomic pathways of the animal steroid hormones are little understood. In this study, we report the participation of a G-protein alpha q (Gαq)1 subunit in the 20E nongenomic pathway in the cell membrane and regulating gene expression during molting and metamorphosis in a lepidopteran insect, Helicoverpa armigera. 20E-induced phosphorylation of Gαq was detected using two-dimensional electrophoresis techniques. Knockdown of Gαq by injecting double-stranded RNA suppressed the development of larvae, delayed metamorphosis, and inhibited 20E-induced gene expression. Gαq was distributed throughout the cell, and migrated toward the plasma membrane upon 20E induction. Gαq was necessary in the 20E-induced intracellular Ca2+ release and extracellular Ca2+ influx. The protein kinase C (PKC) inhibitor could repress 20E-induced phosphorylation of cyclin-dependent kinase 10 (CDK10) and transcription factor ultraspiracle (USP1). PKC inhibitor could repress the Gαq phosphorylation and membrane trafficking. These results suggest that Gαq participates in 20E signaling in the cell membrane at the pre-genomic stage by modulating the increase of the intracellular Ca2+ and phosphorylation of CDK10 and USP1 in 20E transcription complex to regulate gene transcription.