Aim: To determine the effect of long-term inactivation of tyrosine kinases on voltage-gated calcium currents in pancreatic β-cells and to evaluate the function of tyrosine kinases in pancreatic β-cells. Methods: Primarily cultured mouse pancreatic islets and β-cells were pretreated by 0.1 mmol/L genistein for 12 hours. Voltage-gated calcium currents and action potentials were recorded with patch clamp techniques in the configuration of perforated whole-cell recording. RT-PCR method was used to evaluate the changes in expression of voltage-gated calcium channels α1 subunit. Results: After treatment by genistein for 12 hours, the whole-cell voltage-gated calcium currents were significantly diminished(-13.83±1.515 pA/pF vs -7.012±1.502 pA/pF, P<0.01, n=6). The amplitudes of action potentials in genistein-treated β-cells were also significantly attenuated (38.50±7.46 mV vs 15.95±4.39 mV,P<0.01, n=6). The expression of voltage-gated calcium channels α1 subunit in mouse islets was significantly decreased to 0.792±(0.078) of that in control conditions (P<0.01,n=5). Conclusion: Genistein treatment decreases expression and current of voltage-ga-(ted) calcium currents in mouse pancreatic β-cells, which suggests that inhibition of tyrosine kinases activity plays an important role in the dysfunction of pancreatic β-cells.