诺米林对黑色素瘤B16F-10细胞增殖、侵袭和迁移能力的影响

The influence of normirin on the proliferation, invasion and migration of melanoma B16F-10 cells

目的探讨诺米林对黑色素瘤B16F-10细胞增殖、侵袭和迁移能力的影响。方法 CCK-8法检测B16F-10细胞在0～500μmol/L浓度梯度诺米林处理下的细胞活力。B16F-10细胞分为4组:B16F-10对照组,10μmol/L诺米林组,20μmol/L诺米林组和50μmol/L诺米林组。CCK-8法检测细胞增殖,Transwell小室实验检测细胞侵袭,划痕实验检测细胞迁移,Western blotting检测细胞增殖核抗原-67 (Ki-67),血管内皮细胞生长因子(VEGF)和基质金属蛋白酶9 (MMP-9)的表达。结果 <50μmol/L诺米林不会对B16F-10细胞株产生明显的细胞毒性。诺米林组(10、20、50μmol/L)细胞增殖倍数从48 h开始低于对照组(P<0.01)。与对照组比较,诺米林组每个视野下的侵袭细胞数减少(P<0.01),其中对照组、10μmol/L诺米林组、20μmol/L诺米林组、50μmol/L诺米林组的侵袭细胞数分别为(122±15)、(88±20)、(70±13)和(23±9)个。诺米林组划痕愈合率低于对照组(P<0.01),其中对照组、10μmol/L诺米林组、20μmol/L诺米林组、50μmol/L诺米林组的划痕愈合率分别为(95.0±4.7)%、(84.3±7.0)%、(58.2±9.1)%和(36.0±5.9)%。与对照组比较,诺米林组Ki-67,VEGF和MMP-9的蛋白相对表达量降低(P<0.01)。结论诺米林具有抑制黑色素瘤细胞B16F-10的增殖、侵袭和迁移的作用。

Objective To explore the effect of normirin on the proliferation, invasion and migration of melanoma B16F 10 cells. Methods TheactivityofB16F 10 cells treated with 0-500μmol/L concentration of normirin was detected by CCK 8.B16F 10 cells were divided into four groups,including B16F 10 control group,normirin(10μmol/L)group,normirin(20μmol/L) group and normirin (50μmol/L)group.The cell proliferation was detected by CCK 8.Cell invasion was measured by transwell chamber test.Cell migration was analyzed by scratch assay. The expression of antigen identified by monoclonal antibody (Ki 67), vascular endothelial growth factor (VEGF) and matrix metalloprotein 9 (MMP 9) was detected by Western blotting. Results The concentration of normirin less than 50 μmol/L had no obvious cytotoxicity on melanoma cells B16F 10.The cell proliferation in normirin(10,20,50μmol/L)group was lower than that of control group from 48h(P<0 01).Compared with control group,the number of invasive cell sperfieldinnormirin groups de creased(P<0 01),and the invasive numbers of control group,normirin(10,20,50μmol/L)groups were(122±15),(88±20),(70± 13)and(23±9).The wound closure ratein normirin groups was lower than that of control group(P<0 01),and the rates of control, normirin(10,20,50μmol/L)groups were(95 0±4 7)%,(84 3±7 0)%,(58 2±9 1)%and(36 0±5 9)%.Compared with control group,the relative protein levels of Ki 67,VEGFandMMP 9in normirin(10,20,50μmol/L) groups were reduced(P<0 01). Conclusion Normirin can inhibite the proliferation, invasion and migration of melanoma B16F 10 cells.