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Application of the Cameleon Monitoring System of Ca2+ in the Process of H2O2 Induced Apoptosis of A549 Cells
Zhang Siyang1, Yu Miao1, Gao Jian1, Qiu Xueshan2, Cui Zeshi1*
1Center of Laboratory Technology and Experimental Medicine, China Medical University, Shenyang 110001, China; 2Department of Pathology, China Medical University, Shenyang 110001, China
Abstract: The aim was to evaluate the Cameleon monitoring system of Ca2+ in the process of H2O2 induced
the apoptosis of lung adenocarcinoma A549 cells. The cytoplastic Ca2+ concentration ([Ca2+]i) was determined in
real-time， and the correlations between [Ca2+]i and cell apoptosis as well as proline-rich tyrosine kinase 2 (Pyk2)
phosphorylation were explored. The Ca2+ indicator Cameleon YC3.6 was transfected into A549 cells， and cells were
stimulated with 50 mmol/L H2O2 24 h later. Laser scanning confocal microscope was applied to perform real-time
monitoring on the variation of [Ca2+]i in selected cells. Western blot assay was used to evaluate the activation of
Pyk2-tyr402， and DAPI staining was used to observe apoptosis in H2O2 treated cells. Our results showed that the
cytoplastic free [Ca2+]i was rapidly elevated in H2O2 stimulated A549 cells. Pyk2-tyr402 was significantly phosphorylated，
and the apoptotic rate was increased in H2O2 treated cells， compared with untreated ones (P<0.01). In summary，
H2O2 promoted Ca2+ release in A549 cells， and induced cell apoptosis probably by activating Pyk2.
the apoptosis of lung adenocarcinoma A549 cells. The cytoplastic Ca2+ concentration ([Ca2+]i) was determined in
real-time， and the correlations between [Ca2+]i and cell apoptosis as well as proline-rich tyrosine kinase 2 (Pyk2)
phosphorylation were explored. The Ca2+ indicator Cameleon YC3.6 was transfected into A549 cells， and cells were
stimulated with 50 mmol/L H2O2 24 h later. Laser scanning confocal microscope was applied to perform real-time
monitoring on the variation of [Ca2+]i in selected cells. Western blot assay was used to evaluate the activation of
Pyk2-tyr402， and DAPI staining was used to observe apoptosis in H2O2 treated cells. Our results showed that the
cytoplastic free [Ca2+]i was rapidly elevated in H2O2 stimulated A549 cells. Pyk2-tyr402 was significantly phosphorylated，
and the apoptotic rate was increased in H2O2 treated cells， compared with untreated ones (P<0.01). In summary，
H2O2 promoted Ca2+ release in A549 cells， and induced cell apoptosis probably by activating Pyk2.
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