TNF-α诱导小鼠B16黑色素瘤细胞凋亡及其对小鼠B16细胞移植瘤生长的抑制作用

The Effect of TNF-α on the Apoptosis of Murine B16 Melanoma Cells and the B16 cell Xenograft Growth in Mice

目的:探讨TNF-α对小鼠B16黑色素瘤(MM)细胞凋亡的诱导及其对小鼠B16移植瘤生长的抑制作用。方法:用不同浓度的TNF-α直接作用于培养的小鼠B16黑色素瘤细胞,36h后,采用流式细胞仪(FCM)及原位末端标记法(TUNEL法)检测B16细胞的凋亡率。动物实验观察TNF-α小量瘤体内注射对小鼠B16移植瘤生长的抑制作用。结果:在1000、3000、5000及10000U/mLTNF-α作用下,B16细胞的凋亡指数均明显高于空白对照组(P<0.01),随着TNF-α浓度增加,B16凋亡细胞数呈增加趋势。动物实验结果显示,TNF-α治疗3周后,治疗组荷瘤小鼠MM移植瘤的体积和重量明显低于对照组(P<0.01)。结论:TNF-α能够诱导小鼠B16黑色素瘤细胞发生凋亡,且小剂量TNF-α瘤体内注射能显著抑制小鼠移植性MM的生长。

Objective: To investigate the potential role of tumor necrosis factor-α (TNF-α) in inducing apoptosis of murine B16 melanoma cell and in inhibiting the growth of B16 cell Xenograft in C57BL/6 mice. Methods: Cultured murine B16 melanoma cells were treated with TNF-α in different concentration for 36 hours. Flow cytometry and In situ terminal deoxynucleotidual transferase and nick translation assay (TUNEL) were applied to detect the apoptotic rate of different groups of B16 cells. The inhibitive effect of TNF-α on B16 cell xenograft growth was studied in vivo by admistering TNF-α with a small dose every day locally. Results: TNF-α. could significantly induce apoptosis of B16 cells at the concentration of 1 000、3 000、5 000 and 10 000U/mL(P<0.01), and the apoptotic rate of B16 cells was TNF-α concentration dependent. After treated with TNF-α for three weeks, the volume and the weight of tumor in C57BL/6 mice were significantly less than that of controls (P<0.01). Conclusion: TNF-α can induce the apoptosis of murine B16 melanoma cells, and TNF-α injection with a small dose could inhibit significantly the growth of transplanted MM.