重楼皂苷Ⅱ干预后肺癌细胞增殖、迁移和侵袭 能力变化及其与 miR-16-5p 表达的关系.

Objective To investigate the inhibitory effects of polyphyllin Ⅱ on the proliferation, migration, and in‐ vasion of lung cancer cells and their possible mechanism. Methods Lung cancer A549 cells in the logarithmic growth phase were divided into the high concentration group, medium concentration group, low concentration group, and control group. Polyphyllin Ⅱ of 3, 2, 1 µmol/L and the same amount of dimethyl sulfoxide solution were added for 24 h, respec‐ tively. Cell proliferation capacity （represented by cell proliferation activity) was detected by CCK-8. Cell migration ability （expressed as cell mobility) was measured by cell wound healing. Cell invasion ability （represented by the number of cells invaded) was detected by Transwell invasion assay. The expression levels of apoptosis-related proteins ［Cleave-Caspase3, Bax, Bcl-2, matrix metalloproteinase （MMP-2)］ were detected by Western blotting. Real-time fluorescence quantita‐ tive PCR was used to detect the expression of miR-16-5p. A549 cells in logarithmic growth phase were divided into the miR-16-5p inhibitor group, miR-16-5p NC group, and negative control group. Cells in the miR-16-5p inhibitor group and miR-16-5p NC group were transfected with miR-16-5p inhibitor and miR-16-5p inhibitor NC labeled with green fluores‐ cence and then were added with 2 µmol/L polyphyllin Ⅱ solution, respectively. Cells in the negative control group did not receive cell transfection and were only added with the same amount of dimethyl sulfoxide solution. At 24 h after interven‐ tion, the expression of miR-16-5p, apoptosis-related proteins and cell biological behavior were detected by the above meth‐ ods. Results The cell proliferation viability, cell migration rates and the numbers of invasive cells in the high, medium and low concentration groups and the control group sequentially increased, and significant differences were found between groups （all P<0. 05). Compared with the control group, the expression levels of Cleave-Caspase-3, Bax and miR-16-5p in‐ creased in the high, medium and low concentration groups, and the expression levels of Bcl-2 and MMP-2 protein de‐ creased, with the most significant changes in the high concentration group （all P<0. 05). The relative miR-16-5p expres‐ sion levels in the miR-16-5p inhibitor group, miR-16-5p NC group and negative control group were 0. 07 ± 0. 05, 1. 57 ± 0. 07 and 1. 00 ± 0. 03, respectively； the relative miR-16-5p expression was lower in the miR-16-5p inhibitor group than in the miR-16-5p NC group and negative control group （all P<0. 05). Cell proliferation viability, cell migration rates, the numbers of invading cells and the expression levels of Bcl-2 and MMP-2 protein were lower in the miR-16-5p NC group than in the miR-16-5p inhibitor group and negative control group； Cleave-Caspase-3 and Bax protein expression levels were higher than those of the miR-16-5p inhibitor group and negative control group （all P<0. 05)； there were no statistical‐ ly significant differences in the above indicators between the miR-16-5p inhibitor group and the negative control group （all P>0. 05). Conclusion Polyphyllin Ⅱ inhibited the proliferation, migration and invasion of lung cancer cells, and the mechanism may be related to the regulation of miR-16-5p expression. [ABSTRACT FROM AUTHOR]