(−)-Englerin A-evoked Cytotoxicity Is Mediated by Na(+) Influx and Counteracted by Na(+)/K(+)-ATPase

(−)-Englerin A ((−)-EA) has a rapid and potent cytotoxic effect on several types of cancer cell that is mediated by plasma membrane ion channels containing transient receptor potential canonical 4 (TRPC4) protein. Because these channels are Ca(2+)-permeable, it was initially thought that the cytotoxicity arose as a consequence of Ca(2+) overload. Here we show that this is not the case and that the effect of (−)-EA is mediated by a heteromer of TRPC4 and TRPC1 proteins. Both TRPC4 and TRPC1 were required for (−)-EA cytotoxicity; however, although TRPC4 was necessary for the (−)-EA-evoked Ca(2+) elevation, TRPC1 was not. TRPC1 either had no role or was a negative regulator of Ca(2+) entry. By contrast, both TRPC4 and TRPC1 were necessary for monovalent cation entry evoked by (−)-EA, and (−)-EA-evoked cell death was dependent upon entry of the monovalent cation Na(+). We therefore hypothesized that Na(+)/K(+)-ATPase might act protectively by counteracting the Na(+) load resulting from sustained Na(+) entry. Indeed, inhibition of Na(+)/K(+)-ATPase by ouabain potently and strongly increased (−)-EA-evoked cytotoxicity. The data suggest that (−)-EA achieves cancer cell cytotoxicity by inducing sustained Na(+) entry through heteromeric TRPC1/TRPC4 channels and that the cytotoxic effect of (−)-EA can be potentiated by Na(+)/K(+)-ATPase inhibition.