多巴胺对胶质瘤细胞U251增殖、侵袭及VEGF表达影响

Effects of dopamine on proliferation, invasion, and expression of vascular endothelial growth factor in glioma cell line U251

目的探讨多巴胺对胶质瘤细胞U251增殖、侵袭及血管内皮生长因子(VEGF)表达的影响及意义。方法将人胶质瘤细胞株U251分为5组,分别加入多巴胺0(对照)、10、25、50、100μmol/L,分别于培养12、24、36、48h时,采用细胞计数试剂盒(CCK-8)法检测细胞增殖情况,采用流式细胞仪和Transwell实验检测培养48h后细胞周期及侵袭能力变化,采用Westernblot检测细胞中细胞周期蛋白D1(cyclinD1)、细胞周期蛋白依赖性激酶4(CDK-4)、CDK-6及VEGF、基质金属蛋白酶2(MMP-2)、MMP-9表达。结果与对照组[(92.36±1.37)%]比较,多巴胺培养48h时,10、25、50和100μmol/L组U251细胞增殖百分比[(46.82±1.17)%、(41.96±1.06)%、(37.52±0.98)%、(30.57±1.11)%]明显下降(P<0.05),呈剂量依赖性(P<0.05);与对照组比较,多巴胺10、25、50和100μmol/L组U251细胞G0/G1期比例明显增多(P<0.05),S期细胞比例明显减少(P<0.05),具有剂量依赖性;与对照组[(1099.82±33.57)个]比较,多巴胺10、25、50和100μmol/L组U251细胞侵袭数目[分别为(776.52±25.16)、(555.43±22.57)、(442.33±12.03)、(336.82±14.11)个]均明显减少(P<0.05),呈一定剂量依赖性(P<0.05);与对照组比较,多巴胺10、25、50和100μmol/L组U251细胞中cyclinD1、CDK4、CDK6及VEGF、MMP-2、MMP-9蛋白表达量均明显降低(P<0.05)。

Objective To explore effects and significances of dopamine on the proliferation, invasion, and expression of vascular endothelial growth factor (VEGF) in glioma cell line U251.Methods Human glioma cells line U251 were divided into 5 groups administered with dopamine at doses of 0 (control), 10, 25, 50, and 100 μmol/L. CCK-8 method was used to detect cell proliferation at 0, 12, 24, 36 and 48 hours of dopamine treatment. Flow cytometry and Transwell assay were used to detect changes of cell cycle and invasive ability 48 hours after dopamine treatment. The expressions of cyclin D1, cyclin dependent kinase 4 (CDK-4), cyclin dependent kinase 6 (CDK-6), VEGF, matrix metalloproteinase 2 (MMP-2), and matrix metalloproteinase 9 (MMP-9) were detected with Western blot.Results Compared to the control cells, the U251 cells with 48 hours' treatment of dopamine at doses of 10, 25, 50, and 100 μmol/L showed following significant variations in significant dose-dependent manners: decreased cell proliferation percentage (46.82 ±1.17%, 41.96 ±1.06%, 37.52 ±0.98%, and 30.57 ±1.11% vs. 92.36 ±1.37%, P < 0.05 for all);increased percentage of cells in G0/G1 phase (P < 0.05 for all);decreased percentage of cells in S phase (P < 0.05 for all);reduced number of invasive cells (776.52 ±25.16, 555.43 ±22.57, 442.33 ±12.03, and 336.82 ±14.11 vs. 1 099.82 ±33.57, P < 0.05 for all);and decreased intracellular expressions of cyclin D1, CDK4, CDK6, VEGF, MMP-2, and MMP-9 (all P < 0.05).Conclusion Dopamine could inhibit the proliferation and invasion of glioma cell line U251 and the effects may be related to the inhibited intracellular expressions of cyclin D1, CDK-4, CDK-6, VEGF, MMP-2, and MMP-9.