来那度胺通过抑制VEGF蛋白表达抑制人肝癌细胞LM3的迁移和侵袭

目的 研究来那度胺对人肝癌细胞LM3细胞迁移和侵袭能力的影响。方法 采用不同浓度的来那度胺（0、5、10、20、40、80 μmol/L）处理人肝癌细胞LM3，通过检测对LM3细胞活性变化和血管内皮生长因子（VEGF）蛋白表达量变化筛选最适实验药物浓度。通过划痕实验以及Transwell侵袭实验检测来那度胺对LM3细胞迁移率及侵袭数量的影响。采用蛋白印迹法（Western blotting）检测上皮间质转化（epithelial-mesenchymal transition，EMT）相关蛋白标志物E-cadherin、Vimentin、Snail、VEGF的表达情况。结果 确定40 μmol/L来那度胺为最佳浓度。40 μmol/L来那度胺处理LM3细胞48 h后迁移率为（40.75±4.17）%，侵袭数为（43.67±9.03），与对照组迁移率（66.80±3.49）%，侵袭数（135.77±13.67）]比较具有统计学差异（P＜0.05）。在VEGF作用下LM3细胞发生EMT，迁移率（84.66±5.09）%和侵袭数（229.73±15.16）明显增加（均P＜0.05）；同时VEGF表达量也升高（P＜0.05）。而来那度胺作用于VEGF预处理细胞后，VEGF表达量下降（P＜0.05），EMT受到抑制，随之迁移率（57.69±4.63）%和侵袭数（108.63±13.27）降低（P＜0.05）。结论 本研究表明，来那度胺可能通过抑制VEGF蛋白表达从而抑制人肝癌细胞LM3的迁移和侵袭。

Lenalidomide inhibits the migration and invasion of human hepatomacell line LM3 by inhibiting the expression of VEGF

Objective To investigate the effect of lenalidomide on migration and invasion of human hepatoma cell line LM3. Methods LM3 cells were dealt with lenalidomide (0, 5, 10, 20, 40, 80 μmol/L respectively). To select the optimal drug concentration, changes of cell viability and the protein expression of VEGF were tested. The wound healing assay and transwell assay were employed to detect the migration and invasion capability of LM3 cells. Western blotting was used to examine the expression of epithelial-mesenchymal transition (EMT) related molecular-biological factors E-cadherin, Vimentin, Snail, and VEGF. Results The optimal concentration of lenalidomide 40 μmol/L was established in this study. Lenalidomide decreased the migration rate (40.75±4.17)% and invasion number (43.67±9.03) of LM3 cells, with statistical significance compared with those of the control group migration rate (66.80±3.49)%, invasion number (135.77±13.67)] (P<0.05). The migration rate (84.66±5.09)% and invasion number (229.73±15.16) of LM3 cells were significantly increased after activating epithelial-mesenchymal transition (EMT) of LM3 cells by VEGF (P<0.05), meanwhile the expression of VEGF increased (P<0.05). After treatment of VEGF-induced cells with lenalidomide, the expression of VEGF was decreased and EMT was inhibited, thus the migration rate (57.69±4.63)% and invasion number (108.63±13.27) were weakened (P<0.05). Conclusion This study suggests that lenalidomide inhibits the migration and invasion of human hepatoma cell line LM3 by blocking the protein expression of VEGF.