The Dormancy Effect Of The Serum Containing Yiqi Yangyin Decoction On Lung Cancer A549 Cells

Objective: To study the effects of the serum containing Yiqi Yangyin decoction on growth inhibition?cell cycle changes?the expression of PCNA mRNA and protein and the expression of VEGF mRNA and protein of non-small cell lung cancer A549 cell line.To explore the mechanism of the serum containing Yiqi Yangyin decoction on tumor dormancy induction.Methods: Twenty SD rats were randomly divided into 4 groups: control group,high dose group,medium dose group and low dose group,with 5 rats in each group.The Yiqi Yangyin decoction is boiled and concentrated.The medium dose is the equivalent dose,that is,the daily dose of 10.26 g/100 g rat is required;the high dose is multiplied by 1.5,that is,the daily dose is 15.39 g/100 g rats,low dose divided by 1.5,that is,a daily dose of 6.84 g / 100 g rats.The control group was given the same amount of normal saline and was continuously intragastrically administered for 3 days.The eye blood was collected 2 hours after the last administration,and the drug-containing serum was isolated,and the serum collected from the same group of 5 mice was thoroughly mixed.After A549 cells were pre-cultured for 24 h into logarithmic growth phase,the serum containing the control group,high,medium and low dose groups were added to the medium and cultured for 24 h.The cell inhibition rate was detected by CCK-8 method,and the cell cycle was detected by flow cytometry,the expression levels of PCNA and VEGF mRNA were detected by real-time PCR,and the expression levels of PCNA and VEGF protein were detected by Western blotting.Results: 1.Cell proliferation inhibition experiment: High?medium and low dose groups of drug-containing serum were added to the culture medium and acted on A549 cells for 24 hours.CCK-8 method was used to detect the proliferation inhibition effect of drug-containing serum on A549 cells.Compared with the control group,the high ? medium and low dose groups all have inhibitory effects with statistical significance(P(27)0.01).The inhibitory rates of the high,medium and low dose groups are 46.09%,57.45% and 70.27% respectively.2.Cell cycle block experiment: Compared with the control group,the G0/G1 phase cell ratio of A549 cells treated with different doses of drug-containing serum increased,and the difference was statistically significant(P(27)0.05).Cell division cycle stagnated in G0/G1 phase after A549 cells were treated with high?medium and low dose groups.3.Detection of expression level of PCNA mRNA and protein: Compared with the control group,the expression level of PCNA mRNA in the low-dose and medium-dose groups decreased with statistical significance(P(27)0.05).The expression level of PCNA mRNA in the high-dose treatment group increased slightly after the treatment,but the difference was not significant(P(29)0.05).Compared with the control group,the expression of PCNA protein in each dose of drug-containing serum treatment group was decreased,and the difference was statistically significant(P(27)0.01).4.Detection of VEGF mRNA and protein expression level: Compared with the control group,the treatment groups with medium and low doses of the serum containing Yiqi Yangyin decoction had no significant effect on VEGF mRNA expression in A549 cells(P(29)0.05),while the relative expression of VEGF protein in cells of high dose treatment group was increased,and the difference was statistically significant(P(27)0.01).Conclusion:An appropriate concentration of Yiqi Yangyin may inhibit cell proliferation by decreasing the expression of PCNA,and inducing A549 cells to enter cell dormancy.The effect of Yiqi Yangyin decoction on the dormancy of A549 cells may be independent of the down-regulation of angiogenic pathway VEGF factor.