Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2 Induced Apoptosis of A549 Cell
Background and objective Lung cancer is a common malignant tumor all over the world, and Ca2+ is a critical regulator for apoptosis of cancer cells. The monitoring of cytoplastic Ca2+ level in real-time will contribute to further investigate the molecular mechanisms of apoptosis mediated by Ca2+ in...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | zho |
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Chinese Anti-Cancer Association; Chinese Antituberculosis Association
2014-03-01
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| Series: | Chinese Journal of Lung Cancer |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.3779/j.issn.1009-3419.2014.03.03 |
| _version_ | 1818700478869929984 |
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| author | Siyang ZHANG Chunyan LI Jian GAO Xueshan QIU Zeshi CUI |
| author_facet | Siyang ZHANG Chunyan LI Jian GAO Xueshan QIU Zeshi CUI |
| author_sort | Siyang ZHANG |
| collection | DOAJ |
| description | Background and objective Lung cancer is a common malignant tumor all over the world, and Ca2+ is a critical regulator for apoptosis of cancer cells. The monitoring of cytoplastic Ca2+ level in real-time will contribute to further investigate the molecular mechanisms of apoptosis mediated by Ca2+ in lung cancer cells. To evaluate the Ca2+ indicator fluo-3 and fluo-4 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 were investigated. The differences in fluorescence intensity and measured value were compared between the two Ca2+ indicators. Methods Cells were loaded with the Ca2+ indicator fluo-3 or fluo-4 for 1 h, and then stimulated with 50 mM H2O2. Laser scanning confocal microscope was applied to perform real-time monitoring on the variation of [Ca2+]i in selected cells. DAPI staining was used to observe apoptosis in H2O2 treated cells. Results Our results showed that the fluorescence intensity of fluo-4 was stronger than that of fluo-3 in the same condition of dye concentration, loading time and image acquisition parameters before or after H2O2 stimulation. The cytoplastic [Ca2+]i was rapidly elevated in H2O2 stimulated A549 cells. The range of [Ca2+]i in selected cells loaded with fluo-3 was 112.2 nM-1,069.6 nM, and that in selected cells loaded with fluo-4 was 7.6 nM-505.4 nM. Moreover, the apoptotic rate was significantly increased in H2O2 treated cells, compared with untreated ones (P<0.01). Conclusion In summary, H2O2 promoted Ca2+ release in A549 cells, and induced cell apoptosis. Ca2+ indicator fluo-4 was probably more applicable to measure [Ca2+]i in cells with less content of Ca2+. |
| first_indexed | 2024-12-17T15:05:35Z |
| format | Article |
| id | doaj.art-982a3b25344945338603febb5b2d1ad6 |
| institution | Directory Open Access Journal |
| issn | 1009-3419 |
| language | zho |
| last_indexed | 2024-12-17T15:05:35Z |
| publishDate | 2014-03-01 |
| publisher | Chinese Anti-Cancer Association; Chinese Antituberculosis Association |
| record_format | Article |
| series | Chinese Journal of Lung Cancer |
| spelling | doaj.art-982a3b25344945338603febb5b2d1ad62022-12-21T21:43:48ZzhoChinese Anti-Cancer Association; Chinese Antituberculosis AssociationChinese Journal of Lung Cancer1009-34192014-03-0117319720210.3779/j.issn.1009-3419.2014.03.03Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2
Induced Apoptosis of A549 CellSiyang ZHANG0Chunyan LI1Jian GAO2Xueshan QIU3Zeshi CUI4Center of Laboratory Technology and Experimental Medicine, China Medical University,
Shenyang 110001, ChinaCenter of Laboratory Technology and Experimental Medicine, China Medical University,
Shenyang 110001, ChinaCenter of Laboratory Technology and Experimental Medicine, China Medical University,
Shenyang 110001, ChinaDepartment of Pathology, China Medical University,
Shenyang 110001, ChinaCenter of Laboratory Technology and Experimental Medicine, China Medical University,
Shenyang 110001, ChinaBackground and objective Lung cancer is a common malignant tumor all over the world, and Ca2+ is a critical regulator for apoptosis of cancer cells. The monitoring of cytoplastic Ca2+ level in real-time will contribute to further investigate the molecular mechanisms of apoptosis mediated by Ca2+ in lung cancer cells. To evaluate the Ca2+ indicator fluo-3 and fluo-4 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 were investigated. The differences in fluorescence intensity and measured value were compared between the two Ca2+ indicators. Methods Cells were loaded with the Ca2+ indicator fluo-3 or fluo-4 for 1 h, and then stimulated with 50 mM H2O2. Laser scanning confocal microscope was applied to perform real-time monitoring on the variation of [Ca2+]i in selected cells. DAPI staining was used to observe apoptosis in H2O2 treated cells. Results Our results showed that the fluorescence intensity of fluo-4 was stronger than that of fluo-3 in the same condition of dye concentration, loading time and image acquisition parameters before or after H2O2 stimulation. The cytoplastic [Ca2+]i was rapidly elevated in H2O2 stimulated A549 cells. The range of [Ca2+]i in selected cells loaded with fluo-3 was 112.2 nM-1,069.6 nM, and that in selected cells loaded with fluo-4 was 7.6 nM-505.4 nM. Moreover, the apoptotic rate was significantly increased in H2O2 treated cells, compared with untreated ones (P<0.01). Conclusion In summary, H2O2 promoted Ca2+ release in A549 cells, and induced cell apoptosis. Ca2+ indicator fluo-4 was probably more applicable to measure [Ca2+]i in cells with less content of Ca2+.http://dx.doi.org/10.3779/j.issn.1009-3419.2014.03.03Fluo-3Fluo-4Ca2+H2O2Apoptosis |
| spellingShingle | Siyang ZHANG Chunyan LI Jian GAO Xueshan QIU Zeshi CUI Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2 Induced Apoptosis of A549 Cell Chinese Journal of Lung Cancer Fluo-3 Fluo-4 Ca2+ H2O2 Apoptosis |
| title | Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2
Induced Apoptosis of A549 Cell |
| title_full | Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2
Induced Apoptosis of A549 Cell |
| title_fullStr | Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2
Induced Apoptosis of A549 Cell |
| title_full_unstemmed | Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2
Induced Apoptosis of A549 Cell |
| title_short | Application of the Ca2+ Indicator Fluo-3 and Fluo-4 in the Process of H2O2
Induced Apoptosis of A549 Cell |
| title_sort | application of the ca2 indicator fluo 3 and fluo 4 in the process of h2o2
induced apoptosis of a549 cell |
| topic | Fluo-3 Fluo-4 Ca2+ H2O2 Apoptosis |
| url | http://dx.doi.org/10.3779/j.issn.1009-3419.2014.03.03 |
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