Here, we found that Hsp90 inhibitors targeting different sites had different effects on the interaction between Hsp90 and PGK1, which resulted in distinct effects on GSK3β stability, to affect the stemness of BCSCs, suggesting that Hsp90 inhibitors, which maintained GSK3β stability, could be effective in the treatment of breast cancer. The gene discussed is GSK3B; the disease is breast carcinoma.