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目的:探讨重组人血管内皮抑制素(recombinant human endostatin,rh-endostatin)联合达卡巴嗪(Dacarbazine)能否增强抗黑素瘤作用,并初步分析其作用机制。方法:分别采用MTT法和FCM法检测重组人血管内皮抑制素单药及其联合达卡巴嗪对黑素瘤B16F10细胞增殖和凋亡的影响;Western印迹法检测重组人血管内皮抑制素对黑素瘤细胞中细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)磷酸化的影响;建立C57BL/6黑素瘤荷瘤小鼠模型,研究重组人血管内皮抑制素联合达卡巴嗪能否增强体内抗黑素瘤的作用。结果:重组人血管内皮抑制素、达卡巴嗪以及两药联合处理对黑素瘤B16F10细胞增殖的抑制率分别为(9.67±2.89)%、(22.67±3.06)%和(30.33±1.16)%,差异有统计学意义(P<0.05);重组人血管内皮抑制素组黑素瘤细胞的ERK分子磷酸化水平明显降低。接种肿瘤18d时,重组人血管内皮抑制素组、达卡巴嗪组和两药联合组荷瘤小鼠的肿瘤体积分别为(6.45±1.24)cm3、(8.94±2.04)cm3和(4.56±0.98)cm3,两药联合组的肿瘤体积明显低于达卡巴嗪单药组(P<0.05)。结论:重组人血管内皮抑制素联合达卡巴嗪体外可增强抗黑素瘤B16F10细胞增殖的作用,体内可增强达卡巴嗪对荷瘤小鼠黑素瘤生长的抑制作用,并提高荷瘤小鼠的存活率。推测抑制黑素瘤细胞中ERK分子酪氨酸磷酸化可能是重组人血管内皮抑制素发挥其抗黑素瘤作用的机制之一。
Objective: To investigate whether recombinant human endostatin (rh-endostatin) combined with Dacarbazine can enhance the anti-melanoma effect and to analyze its mechanism. Methods: MTT assay and FCM assay were used to detect the effect of recombinant human endostatin monotherapy combined with dacarbazine on the proliferation and apoptosis of melanoma B16F10 cells. Western blotting was used to detect the effect of recombinant human endostatin on melanoma To study the effect of recombinant human endostatin combined with dacarbazine on the phosphorylation of extracellular signal-regulated kinase (ERK) in C57BL / 6 melanoma-bearing mice. Anti-melanoma effect. Results: The inhibitory rates of recombinant human endostatin, dacarbazine and the combination of the two drugs on the proliferation of melanoma B16F10 cells were (9.67 ± 2.89)%, (22.67 ± 3.06)% and (30.33 ± 1.16)%, respectively (P <0.05). The phosphorylation level of ERK in melanoma cells of recombinant human endostatin group was significantly decreased. On the 18th day after tumor inoculation, the tumor volumes in the recombinant human endostatin group, the dacarbazine group and the two drug-bearing tumor-bearing mice were (6.45 ± 1.24) cm3, (8.94 ± 2.04) cm3 and (4.56 ± 0.98) cm3. The tumor volume of the combination group was significantly lower than that of the dacarbazine single group (P <0.05). CONCLUSION: Recombinant human endostatin combined with dacarbazine can enhance the proliferation of anti-melanoma B16F10 cells in vitro, which can enhance the inhibitory effect of dacarbazine on the growth of melanoma in mice and increase the tumor-bearing mice Survival rate. Presumably inhibition of ERK tyrosine phosphorylation in melanoma cells may be one of the mechanisms by which human recombinant human endostatin exerts its anti-melanoma effect.