Dataset for: TRPM7-mediated spontaneous Ca2+ entry regulates the proliferation and differentiation of human leukaemia cell line K562

AIM: Continuous Ca2+ influx is essential to maintain intracellular Ca2+ homeostasis and its dysregulation leads to a variety of cellular dysfunctions. In this study, we explored the functional roles of spontaneous Ca2+ influx for the proliferation and differentiation of a human erythromyeloid leukaemia cell line K562. METHODS: mRNA/protein expressions were assessed by the real-time RT-PCR, Western blotting and immunocytochemical staining. Intracellular Ca2+ concentration ([Ca2+]i) and ionic currents were measured by fluorescent imaging and patch clamping techniques, respectively. Cell counting/viability and colorimetric assays were applied to assess proliferation rate and haemoglobin synthesis, respectively. RESULTS: Elimination of extracellular Ca2+ decreased basal [Ca2+]i in proliferating K562 cells. Cation channel blockers such as SK&F96365, 2-APB, Gd3+ and FTY720 dose-dependently decreased basal [Ca2+]i. A spontaneously active inward current (Ispont) contributive to basal [Ca2+]i was identified by the nystatin-perforated whole-cell recording. Ispont permeated Ca2+ comparably to Na+, and was greatly eliminated by siRNA targeting TRPM7, a melastatin member of the transient receptor potential (TRP) superfamily. Consistent with these findings, TRPM7 immune-reactivity was detected by Western blotting, and immunofluorescence representing TRPM7 was found localized to the K562 cell membrane. Strikingly, all these procedures, i.e. Ca2+ removal, TRPM7 blockers and siRNA-mediated TRPM7 knockdown significantly retarded the growth and suppressed hemin-induced γ-globin and haemoglobin syntheses in K562 cells, respectively, both of which appeared associated with the inhibition of ERK activation. CONCLUSIONS: These results collectively suggest that spontaneous Ca2+ influx through constitutively active TRPM7 channels may critically regulate both proliferative and erythroid differentiation potentials of K562 cells.