Involvement of Connexin43 hemichannel in ATP release after -irradiation

Ohshima, Yasuhiro; Tsukimoto, Mitsutoshi*; Harada, Hitoshi*; Kojima, Shuji*

We have recently reported that -irradiation induces ATP release from B16 melanoma cells, which is dependent on P2X receptor. However, the mechanism of ATP release caused by irradiation remains unclear. We here show the involvement of Cx43 in P2X receptor-dependent ATP release after 0.5 Gy -irradiation. Inhibitors of gap junction hemichannels and an inhibitory peptide for Cx43 (gap26), but not an inhibitory peptide for pannexin1 (Panx1), significantly blocked -irradiation-induced ATP release from B16 melanoma cells. We confirmed high expression of Cx43 mRNA in B16 melanoma cells. These results suggest involvement of Cx43 in the radiation-induced ATP release. We found that tyrosine phosphorylation after 0.5 Gy -irradiation was significantly blocked by P2X receptor antagonist, but not gap26, suggesting that tyrosine phosphorylation is a downstream event from P2X receptor. Since tyrosine kinase inhibitor significantly suppressed radiation-induced ATP release, tyrosine phosphorylation appears to play an important role in the Cx43-mediated ATP release downstream of P2X receptor. In conclusion, the Cx43 hemichannel, which lies downstream of P2X receptor, is involved in ATP release in response to radiation.