Hydrogen-Induced Apoptosis And Proliferation Inhibition In Lung Adenocarcinoma A549 Cells And Its Mechanism

Objective:1.To investigate the role of hydrogen in promoting apoptosis and inhibiting the proliferation of lung adenocarcinoma A549 cells and its mechanism.2.The expression levels of P21,CDK4,caspase-7and caspase-9 were identified by western blot in control group,20% hydrogen group,40% hydrogen group and 60% hydrogen group.3.Immunohistochemical experiments verified whether there were differences in the expression of P21,CDK4,caspase-7,and caspase-9 in tumor tissues in the control group,hydrogen group and in the cispltain group.Methods:1.Human lung cancer A549 cells were cultured to logarithmic growth phase,and then were intervened with 20%,40% and 60% hydrogen respectively.Each intervention lasted for 2 hours and lasted for 3 days.The growth of cells was observed under a microscope,the images were collected before intervention every day.Cells were collected after the intervention.Cell apoptosis and cell cycle were checked up by the flow cytometry.2.Conventional cultured adenocarcinoma A549 cells were used as the control group.Adenocarcinoma A549 cells treated according to the first step were used as the experience group.After treatment,the protein was collected and extracted.The expression of P21,CDK4,caspase-7 and caspase-9 was detected by Western blot.3.The expression of P21,CDK4,caspase-7,and caspase-9 in tumor tissues was detected by immunohistochemistry.Results:1.20%,40% and 60% hydrogen can promote the apoptosis and inhibit the proliferation of lung cancer A549 cells.2.The expression of P21 in A549 cells treated with 40% hydrogen and 60% hydrogen showed an upward trend compared with the control group.The expression of CDK4 in A549 cells treated with 60% hydrogen showed a downward trend compared with the control group.The expression of caspase 7 in A549 cells treated with 20% hydrogen,40% hydrogen and 60% hydrogen showed an upward trend compared with the control group.The expression of caspase 9 in A549 cells treated with 40% hydrogen and 60% hydrogen showed an upward trend compared with the control group.The statistical data showed that the difference was statistically significant(P < 0.05).The expression of P21 in the control group and 20% hydrogen group showed no statistical significance(P > 0.05).The expression of CDK4 in control group,20% hydrogen group and 40% hydrogen group had no statistical difference(P > 0.05),There was no significant difference in the expression of caspase 9 between the control group and the 20% hydrogen group(P > 0.05).3.We used the immunohistochemistry to detect the expression of P21,CDK4,caspase 7,caspase9 in tumor tissues.The results showed that the expression of P21,caspase7,caspase9 in the hydrogen and cisplatin groups was higher than expression of P21,caspase7,caspase9 in the control group.We used the immunohistochemistry to detect the expression of CDK4 as the same.The results showed that the expression of CDK4 in the hydrogen and in the cisplatin group was lower than that in the control group.The difference between them had significant(P<0.05).Conclusion:1.High concentration of hydrogen can promote the apoptosis of lung cancer A549 cells and inhibit the proliferation of lung cancer A549 cells.2.High concentration of hydrogen could increase the expression of P21,caspase7,caspase9 protein and decrease the expression of CDK4 protein in tumor tissues and lung cancer A549 cell lines.It is suggested that hydrogen may play a role in promoting apoptosis and inhibiting proliferation of lung cancer A549 cells by increasing the expression of P21,caspase7,caspase9 protein and decreasing the expression of CDK4 protein.