To explore the relationship between Hsc70 and Hsp90 with ERR, 20E primary and secondary response genes, and how do the ERR and 20E-related genes involved in the 20E signaling pathway and male fertility, adult male crickets Teleogryllus emma Ohmschi & Matsumura were divided into the experimental group, negative group, and control group. Crickets of the experimental group were injected with dsTeHsc70 and dsTeHsp90, and the negative group received EGFP-dsRNA, then the efficiency of TeHsc70- and TeHsp90-RNAi and expression of TeERR and 20E-related genes were detected in the experimental group after the crickets treated for 48, 72 and 96 h. Results showed that the TeEcR, TeERR, and TeUsp were highly expressed in free-treated adult testes, and TeERR expression was significantly higher than TeEcR and TeUsp (P<0.05). In addition, expressions of the TeHsc70 and TeHsp90 mRNA is significantly down-regulated(P<0.05) after the crickets treated for 96 h using 2 000 ng dsTeHsc70+dsTeHsp90. Furthermore, expression of the TeERR and 20E primary and secondary response genes, TeEcR, TeUsp, TeE75B, and TeHR3 mRNA, could also be significantly down-regulated (P<0.01). All these results indicate that Hsc70 and Hsp90 are involved in the 20E signal transduction pathway and maintain the development and morphogenesis of testis in crickets T. emma.