Our present study utilized the two-electrode voltage clamp in Xenopus oocyte, Ca imaging in cultured breast cancer cells MCF-7 and MDA-MB-231, molecular biology, and molecular dynamics (MD) simulation to investigate a functionally autoinhibitory intramolecular interaction between TRPV6 helices S5 and S6 and how GOF of TRPV6 and pathogenic mutation R532Q contribute to breast cancer cell progression through a PI3K/Akt/GSK-3β pathway. Here, TRPV6 is linked to breast cancer.