| Abstract: Objective: To investigate the possible effects and mechanisms about compound radix sophora flavescentis injection on proliferation and the expression of heparanase(Hpa) of human lung adenocarcinoma cell line A549 in vitro. Methods: (1) Culture of A549 cells: A549 cell line was inoculated in 25ml culture bottle of RPMI 1640 medium which containing 10% neonatal calf serum in the 5%CO2, 37°C incubator. (2) A549 cells of logarithmic period were used in this study. Time period was divided into three according to the treatment of the compound radix sophora flavescentis injection in A549 cells. Each time period was divided into 15 groups by the drug concentration: 0.02mg/ml(Group2), 0.04mg/ml(Group3), 0.06mg/mI(Group4), 0.08mg/ml(Group5), O.lmg/ml (Group6), 0.2mg/ml(Group7), 0.3mg/ml(Group8), 0.4mg/ml(Group9), 0.5mg/ml(Group l0), 1.0mg/ml(Group 11), 1.5mg/ml(Groupl2), 2.0mg/ml (Groupl3), 2.5mg/ml(Groupl4), control group(Groupl), and 6 holes in each group. The absorbance was detected by WST-1 in all groups after administration of compound radix sophora flavescentis injection, and calculated their cell growth inhibition rates. (3) The expressions of Hpa were dectected by immunocytochemical SP(streptavidin-peroxidase)assay in 1 and 6-14 groups in each time period and compared between the experimental groups and the control group. (4) The invasion ability of these cells were dectected by in 1 and 6-14 groups in each time period and compared between the experimental groups and the control group. Results: (1) The absorbances of A549 cells were decreased after administration of the compound radix sophora flavescentis injection for 24 hours. The control group compared with the experimental groups (Group2 to Group7 and Group 11 to Group 14) were statistically significant (P<0.05).The proliferation of cells could be inhibited under these concentration of the compound radix sophora flavescentis injection in the experimental groups(Group2 to Group7 and Group 11 to Group 14), but could not in the other groups(Group8 to GrouplO) (P>0.05). There were significant differences in the adjacent experimental groups (Groupll to Groupl4) (P<0.05), and no significant differences in the adjacent experimental groups(Group2 to Group7and Group8 to GrouplO) (P>0.05). In summary, it would appear a first suppressed when the concentration of the experimental group increased, and then halted, but finally appeared again, furthermore the inhibition became more obviously at a certain concentration. (2) The role of compound radix sophora flavescentis injection in the A549 cell after 48 hours. The control group compared with the experimental groups(Group4 to Group5 and Group6 to Group 14) were statistically significant (P<0.05), and were not statistically significant in the comparisons of the control group and the experimental groups(Group2 to Group3) (P>0.05). The result suggested the proliferation of cells could be inhibited at this concentration of the compound radix sophora flavescentis injection. There were significant differences in the Group5 and the experimental groups(Group2 to Group3) (P<0.05). The results suggested the proliferation of A549 cells couldn't be inhibited at these concentrations of the drug. There were differences in the neighboring groups(Group4 and Group6 to Groupl4, Group5 and Group2 to Group3) (P<0.05). Thus, the proliferation of cells could be inhibited when the compound radix sophora flavescentis injection reached a certain concentration, and the inhibition would become more obviously with the drug concentration increasing. (3) The role of compound radix sophora flavescentis injection in the A549 cell after 72 hours. The control group respectively compared with the experimental groups(Group7 to Groupl4) were statistically significant (P<0.05). The results suggested the proliferation of cells could be inhibited at this concentration of the compound radix sophora flavescentis injection. (4) The expression of Hpa was decreased in the nucleus of the A549 cell affected by the compound radix sophora flavescentis injection after 24 hours. The control group respectively compared with the experimental groups(Group 10 to Groupl2) was statistically significant (P<0.05) , but no difference in the experimental groups(Group6 to Group9 and Group 13 to Group 14)and the adjacent experimental groups(Group 10 to Group 12 and Group6 to Groupl4) (P>0.05). The expression of heparanase in these cells could be inhibited at a certain concentration of the drug. (5) The expression of Hpa wasdecreased in the nucleus of the A549 cell affected by the compound radix sophora flavescentis injection after 48 hours. The comparisons in the control group and the experimental groups(Group6 to Group 10) and the adjacent experimental groups(Group6, Group7 and Group8 to Group 10) were statistically significant (P<0.05), but were not statistically significant in the adjacent experimental groups(Group8 to Group 10 and Group6 to Group7). The expression of heparanase in these cells could be inhibited at a certain concentration of the drug. (6) The expression of Hpa was decreased in the nucleus of the A549 cell affected by the compound radix sophora flavescentis injection after 72 hours. The comparisons in the control group and the experimental groups(Group6 to Group7) and the adjacent experimental groups(Group6, Group? and Group8 to GrouplO) were statistically significant (P<0.05), but not statistically significant in the control group and experimental groups(Group8 to Group 10) and the adjacent experimental groups(Group6 and Group7) (P>0.05). The expression of heparanase in these cells could be inhibited at a low concentration of the drug. (7) The number of A549 in migration was decreased under the compound radix sophora flavescentis injection after 24 hours used the Transwell chambers assay. The control group and experimental groups(Group 10 to Group 12) were statistically significant (P < 0.05), and not statistically significant in the other groups (.P>0.05). The comparisons of adjacent experimental groups(Group 10 to Group 12 and Group6 to Group9 and Group 13 to Group 14) were statistically significant (P < 0.05). The migration of A549 could be inhibited at a certain concentration of the drug. (8) The number of A549 in migration was decreased under the compoundradix sophora flavescentis injection after 48 hours used the Transwell chambers assay. The control group respectively compared with experimental groups(Group7 to GrouplO) was statistically significant (P<0.05), and opposite result with Group6 (P>0.05). The Group7 compared with experimental groups(Group8 to GrouplO) were statistically significant, and were not significant in the adjacent experimental groups(Group8 to GrouplO) (P > 0.05). This result suggested the number of A549 migration could be inhibited at a certain concentration of the drug. (9) The number of A549 in migration was decreased under the compound radix sophora flavescentis injection after 72 hours used the Transwell chambers assay. The control group compared with experimental groups(Group6 to Group7) were statistically significant (P<0.05), the same as to the comparisons of djacent experimental groups(Group6 to Group7 and Group8 to GrouplO) (P<0.05). This result suggested that the number of A549 migration could be inhibited at a low concentration of the drug. Conclusion: (1)The proliferation of lung adenocarcinoma A549 cells could be inhibited by the compound radix sophora flavescentis injection, and the inhibition intensity was related to its time and concentration. (2) The inhibition of Compound radix sophora flavescentis injection in the lung adenocarcinoma A549 cells might not completely through the inhibition of the expression of Hpa. (3) The ability of invasion and metastasis in lung adenocarcinoma A549 cell related to the expression of Hpa.
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