目的:研究胰岛素对人胰腺癌细胞侵袭与迁移的作用及其可能机制。方法 :RT-qPCR、Western blot检测胰腺癌细胞株胰岛素受体(insulin receptor,IR)的表达。CCK8检测不同浓度胰岛素刺激下胰腺癌细胞的活性并确定其最适浓度;适宜浓度胰岛素刺激下,CCK8检测胰腺癌细胞增殖能力的改变;通过Transwell实验,检测胰岛素刺激条件下胰腺癌细胞侵袭与迁移能力的变化;Western blot检验胰岛素刺激后胰腺癌细胞基质金属蛋白酶(matrix metalloprotease,MMP)-2、7、9的表达变化。结果:胰腺癌细胞株PANC-1和Miapaca-2的IR表达相对高(P〈0.05);胰岛素浓度为100 nmol/L时,对上述两细胞活性影响最大(P〈0.05);胰岛素能明显促进PANC-1和Miapaca-2的增殖、迁移与侵袭(P〈0.05);胰岛素刺激后胰腺癌细胞迁移与侵袭的改变可能与其MMP-2、7、9的表达增加相关(P〈0.05)。结论:胰岛素可能通过增强MMP-2、7、9表达促进胰腺癌细胞株PANC-1和Miapaca-2的迁移及侵袭。
Objective:To explore the effect of insulin on the pancreatic cancer cell lines and the potential mechanisms.Methods:The expression of insulin receptor (IR) was evaluated by q RT-PCR and Western blot.We examined the effect of insulin with different concentrations on the viability of pancreatic cancer cell lines to choose optimal concentration.After insulin treatment,the viability of pancreatic cancer cells was measured by CCK-8 assay; Invasion and migration of cells were measured by a Transwell chamber assay;Western blot was used to observe the expression of matrix metalloprotease (MMP)-2,MMP-7 and MMP-9.Results: Pancreatic cancer cell lines PANC-1 and Miapaca-2 showed relatively higher expression of IR (P0.05).The cell lines had significantly enhanced viability (P0.05) when treated with insulin of 100 nmol/L.CCK-8 assay showed that the proliferation of the cell lines was increased significantly.Transwell chamber assay showed that insulin significantly enhanced the invasion and migration of pancreatic cancer cells (P 0.05).Western blot proved that the expression of MMP-2,7 and 9 was remarkably increased.Conclusion: Insulin can enhance the invasion and migration of pancreatic cancer cell lines by up-regulating MMP-2,7 and 9.