新型精胺氧化酶抑制剂SI-4650对人恶性黑素瘤A375细胞侵袭和迁移的抑制作用研究

Inhibitory effect of a novel spermine oxidase inhibitor SI-4650 on invasion and migration of human malignant melanoma A375 cells

目的研究新型精胺氧化酶抑制剂-SI-4650对人恶性黑素瘤A375细胞侵袭迁移的影响及其作用机制。方法体外培养A375细胞,将A375细胞分为3组:空白对照组(正常培养)和低、高2个浓度实验组(SI-4650:40,80μmol·L-1),均培养48 h。Transwell法检测细胞迁移和侵袭能力,蛋白质印迹法检测上皮-钙黏素(E-cadherin)、神经-钙黏素(N-cadherin)和基质金属蛋白酶-2(MMP-2)的表达水平。结果空白对照组与低、高2个浓度实验组的迁移数分别为268±35,86±26和32±19;这3组的侵袭穿膜细胞数分别为234±28,32±15和9±5;这3组的E-cadherin蛋白相对表达量分别为1.00±0.06,1.55±0.33和3.34±0.50;这3组的N-cadherin蛋白相对表达量分别为1.00±0.04,0.42±0.06和0.15±0.03;这3组的MMP2蛋白相对表达量分别为0.74±0.03,0.63±0.04和0.34±0.01。上述指标:低、高2个浓度实验组与空白对照组相比,差异均有统计学意义(均P<0.001)。结论 SI-4650可高效抑制人恶性黑素瘤A375细胞迁移和侵袭,其机制可能与影响A375细胞上皮-间质转化进程相关。

Objective To investigate the effects of a novel small molecule inhibitor-- SI-4650 screened out by aiming at spermine oxidase(SMO) on the influence of invasion and metastasis of human malignant melanoma A375 cells and its molecular mechanism.Methods A375 cells were divided into 3 groups: blank control group(normal culture), experimental low dose, high dose gyoups(40,80 μmol ·L-1 SI-4650).They were cultured for 48 h.The cell invasion and migration ability were detected by Transwell.The protein expression levels of E-cadherin,N-cadherin,Vimentin,and matrix metalloproteinase-2(MMP-2) were detected by Western blot.Results The number of migration in blank control group and experimental-L,experimental-H groups were 268±35,86±26,32±19;the invasion transmembrance cells in the three groups were 234±28,32±15,9±5;the relative expression of E-cadherin protein in the three groups were 1.00±0.06,1.55±0.33,3.34±0.50;the relative expression of N-cadherin protein in the three groups were 1.00±0.04,0.42±0.06,0.15±0.03;the relative expression of MMP-2 protein in the three groups were 0.74±0.03,0.63±0.04,0.34±0.01.Comparing between two concentration experimental groups and blank control group,the differences of the factors were significant(all P < 0.001).Conclusion The SI-4650 can inhibit the migration and invasion of human malignant melanoma A375 cells,and its mechanism may be related to epithelial to mesenchymal transition.