Caffeine induced Ca2+ release and capacitative Ca2+ entry in human embryonic kidney (HEK293) cells.

The potential role of endogenous ryanodine receptor (RyR) in modulating Ca2+ handling in HEK293 cells is controversial. Using Fura2/AM, here we provide evidence that caffeine can induce Ca2+ release from inositol 1,4,5-trisphosphate receptor-sensitive stores and Ca2+ entry in early passage numbers of HEK293 cells, but not in late passage ones. Ryanodine blocks caffeine-mediated effect, whereas 4-chloro-m-cresol can mimic these effects. In contrast, an increase in cyclic AMP or activation of voltage-dependent Ca2+ channels does not induce detectable alteration in intracellular Ca2+. Importantly, immunoblotting and staining have revealed that endogenous RyR expression is more abundant in the early than in the late passage cells. Additionally, similar to carbachol, Ca2+ entry in response to caffeine is blocked by capacitative Ca2+ entry inhibitors. These results indicate that the endogenous RyR in HEK293 cells can function as Ca2+ release channels and mediate capacitative Ca2+ entry, but they may be reduced due to cell passage.