To explore the role of Na+/H+ exchanger-1 (NHE-1 ) gene expression in intracellular PH (pHi) regulation and the modulation of apoptosis of human lung cancer cells. Methods: After the human lung cell line A549 cells we...To explore the role of Na+/H+ exchanger-1 (NHE-1 ) gene expression in intracellular PH (pHi) regulation and the modulation of apoptosis of human lung cancer cells. Methods: After the human lung cell line A549 cells were incubated with a 21-mer antisense phosphorothioate oligodeoxynucleotide (ASODN) complementary to NHE-1 gene, the acidification effects of AS-ODN on A549 cells was investigated with fluorescent probe labeling and the apoptotic rate of these cells determined with in situ cell apoptosis detection. Results: Intracellular acidification (pHi from 7. 12 to 6. 55) and cell apoptosis of A549 cells induced by AS-ODN were correlated to dose of the agent. The apoptotic rate was found to be 30. 1 %, 91. 2% and 99. 6% respectively at the end of incubation time of 24, 48 and 72 h with 20μmol/L AS--ODN. Under the electron microscope, morphological changes of cell apoptosis were seen. Conclusion: Intracellular acidification (pHi<6. 55) can trigger apoptosis of human lung cancer cells and NHE-1 gene expression plays an important role in the regulation of apoptosis of the human lung cancer cells through preventing intracellular acidification.展开更多
文摘To explore the role of Na+/H+ exchanger-1 (NHE-1 ) gene expression in intracellular PH (pHi) regulation and the modulation of apoptosis of human lung cancer cells. Methods: After the human lung cell line A549 cells were incubated with a 21-mer antisense phosphorothioate oligodeoxynucleotide (ASODN) complementary to NHE-1 gene, the acidification effects of AS-ODN on A549 cells was investigated with fluorescent probe labeling and the apoptotic rate of these cells determined with in situ cell apoptosis detection. Results: Intracellular acidification (pHi from 7. 12 to 6. 55) and cell apoptosis of A549 cells induced by AS-ODN were correlated to dose of the agent. The apoptotic rate was found to be 30. 1 %, 91. 2% and 99. 6% respectively at the end of incubation time of 24, 48 and 72 h with 20μmol/L AS--ODN. Under the electron microscope, morphological changes of cell apoptosis were seen. Conclusion: Intracellular acidification (pHi<6. 55) can trigger apoptosis of human lung cancer cells and NHE-1 gene expression plays an important role in the regulation of apoptosis of the human lung cancer cells through preventing intracellular acidification.