The Effect Of Citreoviridin On Proliferation And Apoptosis Of Lung Cells

ObjectivesTo observe the effect of citreoviridin(CIT)on the proliferation and apoptosis of lung cancer cells(A549).Methods1.A549 lung cancer cells were cultured in 1640 medium with 10% fetal bovine serum.When growing in a logarithmic phase,the cells were randomLy divided into the following groups: control group(0 mg/L CIT),low-dosage group(0.3 mg/L CIT),and high-dosage group(0.6 mg/L CIT).Then the cells were treated by citreoviridin according to the above dosages of CIT.After being treated for 24 h,48h,and 72 h by CIT,the cells were collected,and MTT method was applied to detect proliferation of each group of cells.The results between different groups were compared,and selected the best time as the PI labeled time according to the results.2.A549 lung cancer cells were cultured and stratified as mentioned above.Accorded the previous step to select the PI labeled time.the cells were collected,and fluorescein PI labeling method and flow cytometry(FCM)assay were used to detect the cell cycle distribution of A549 cells in each group,then further comparison was complemented.3.A549 lung cancer cells were cultured and stratified as mentioned above.After being treated for 24 h and 48 h,the cells were collected,and Annexin V-FITC/PI double staining method and FCM assay were performed to detect the apoptosis rates of each group,and further comparison between different groups was complemented.Results1.The MTT assay showed that the survival rates after 24 h CIT-treatment of CIT were(1.000±0.076%),(0.814±0.104%),and(0.753±0.097%)in control group,low-dosage group,and high-dosage group respectively,the low dose group and high dose group was lower than the control group(P<0.05);The survival rates of 48 h CIT-treatment were(1.000±0.010%),(0.840±0.045)%,and(0.783±0.043%)in control group,low-dosage group,and high-dosage group,respectively,the low dose group and high dose group was lower than the control group(P<0.05),and the high dose group was lower than the low dose group(P<0.05);The survival rates of 72 h CIT-treatment were(1.000±0.066%),(0.890±0.054%),and(0.809±0.074%)in control group,low-dosage group,and high-dosage group,respectively,the low dose group and high dose group was lower than the control group(P<0.05),and the high dose group was lower than the low dose group(P<0.05);selected the 24 h as the PI labeled time according to the results.2.The results of PI labeling and FCM detection showed that the G0/G1 stage after 24 h CIT-treatment were: the control group(62.27±1.72%),low dose group(68.93±1.03%),high dose group(73.92±1.81%),the low dose group and high dose group was higher than the control group(P<0.05),and the high dose group higher than the low dose group(P<0.05);the S stage after 24 h CIT-treatment were: the control group(8.41±0.58%),low dose group(7.30±0.83%),high dose group(5.09±1.95%),the high dose group lower than the control group(P<0.05);the G2 stage after 24 h CIT-treatment were: the control group(29.32±1.39%),low dose group(23.77±1.19%),high dose group(20.99±2.90%),the low dose group and high dose group was lower than the control group(P<0.05),and the high dose group was lower than the low dose group(P<0.05).3.Annexin V-FITC/PI staining method and FCM assay illustrated that the early apoptosis rate after 24 h CIT-treatment were: the control group(1.57±0.39%),low dose group(4.27±0.76%),high dose group(5.66±0.53%),the low dose group and high dose group was higher than the control group(P<0.05),and the high dose group was higher than the low dose group(P <0.05;the late apoptosis rate after 24 h CIT-treatment were: the control group(2.19±044%),low dose group(3.19±0.83%),high dose group(4.74±2.66%),there were no significant difference between the groups;the early apoptosis rate after 48 h CIT-treatment were(3.76±0.81%),low dose group(7.46±0.23%),high dose group(10.40±2.5%),the low dose group and high dose group was higher than the control group(P<0.05);the late apoptosis rate after 48 h CIT-treatment were: the control group(3.75±1.21%),low dose group(5.01±1.59%),high dose group(6.77±1.68%),the low dose group and high dose group was higher than the control group(P<0.05),and the high dose group was higher than the low dose(P<0.05);the 24 h total apoptosis rate compared with the total 48 h apoptosis rate: the total 48 h apoptosis rate was higher than the 24 h total apoptosis rate in low dose groups(P<0.05).the total 48 h apoptosis was not significantly different with the 24 h total apoptosis rate in high dose groups(P>0.05).Conclusions1.CIT inhibits the proliferation of A549 lung cancer cells and the strength increased with CIT levels.2.CIT inhibits A549 lung cell proliferation by blocking cell cycle in the G0/G1 phase.3.CIT may induce the apoptosis of lung cancer cells,the strength increased with CIT levels.