摘要
目的探讨神经纤毛蛋白-1(NRP-1)在皮肤黑色素瘤中细胞株和肿瘤组织中的表达及NRP-1单克隆抗体对皮肤黑色素瘤中细胞株增殖和凋亡的影响。方法 Western blot检测各细的NRP-1表达情况。免疫组化法检测NRP-1蛋白在人正常皮肤组织和皮肤黑色素瘤中的表达;CCK-8试剂盒检测NRP-1单克隆抗体对SK-MEL-1细胞株增殖的影响;流式细胞术检测NRP-1单克隆抗体对SK-MEL-1细胞株凋亡的影响。结果 Western blot检测结果显示,NRP-1在4种人黑色素瘤细胞株中的表达量均明显高于人正常皮肤黑色素细胞株(P<0.05)。免疫组化结果提示,NRP-1在6例正常皮肤组织平均表达指数为0.33±0.52;12例皮肤黑色素瘤组织平均表达指数为7.32±1.68(P <0.05)。细胞增殖实验结果提示,低、中、高浓度组的细胞相对增殖活性均<100%,即低、中、高浓度的NRP-1单克隆抗体均对细胞增殖有抑制作用。流式细胞术结果提示PBS组的细胞凋亡率为(1.43±0.15)%,低浓度组细胞凋亡率为(5.57±1.44)%;终浓度组细胞凋亡率为(13.86±3.84)%,高浓度组细胞凋亡率为(36.03±7.51)%。抗体处理组的细胞凋亡率均较PBS组高(P<0.05)。结论 NRP-1在人皮肤黑色素瘤细胞株和肿瘤组织中均有高表达。NRP-1单克隆抗体可显著抑制人皮肤黑色素瘤细胞株的增殖和促进细胞凋亡。
Objective To explore the expression of neuropilin-1(NRP-1) in cell lines and tumor tissues in cutaneous melanoma and the effect of NRP-1 monoclonal antibody on the proliferation and apoptosis of cell lines in cutaneous melanoma.Methods Western blot test was carried out to detect the expression of NRP-1 in each cell.Immunohistochemical method was used to detect the expression of NRP-1 protein in human normal skin tissues and cutaneous melanoma.CCK-8 kit was used to detect the effect of NRP-1 monoclonal antibody on the proliferation of SK-MEL-1 cell line.Flow cytometry was performed to detect the effect of NRP-1 monoclonal antibody on the apoptosis of SK-MEL-1 cell line.Results Western blot results showed that the expression level of NRP-1 in four human melanoma cell lines was significantly higher than that of the human normal skin melanin cell lines(P <0.05).The results of immunohistochemistry showed that the average expression index of NRP-1 was 0.33±0.52 in 6 cases with normal skin tissues and 7.32±1.68 in 12 cases with cutaneous melanoma tissues(P <0.05).The results of cell proliferation experiments suggested that the relative proliferative activity of cells in the low,medium,and high concentration groups was less than 100%,that is,the lowmedium-and high-concentration NRP-1 monoclonal antibodies had an inhibitory effect on cell proliferation.The results of flow cytometry indicated that the apoptosis rate was(1.43±0.15)% for the PBS group,(5.57±1.44)% for the low-concentration group,(13.86±3.84)% for the final concentration group,and(36.03±7.51)% for the high concentration group.The apoptosis rate of the antibody treatment group was higher than that of the PBS group(P <0.05).Conclusion NRP-1 is highly expressed in human cutaneous melanoma cell lines and tumor tissues.NRP-1 monoclonal antibody can significantly inhibit the proliferation of human cutaneous melanoma cell lines and promote cell apoptosis.
作者
洪小兰
吴海丽
谢彩丽
沈翠蓉
钱江
Hong Xiaolan;Wu Haili;Xie Caili;Shen Cuirong;Qian Jiang(Hospital 909 of PLA(The Affiliated Dongnan Hospital of Xiamen University),Zhangzhou,Fujian,363000,China;Chinese Pexple's Liberation Army Xiamen Special Service Convalescen Center,Xiamen,Fujian,361000,China)
出处
《西南国防医药》
CAS
2021年第4期310-314,共5页
Medical Journal of National Defending Forces in Southwest China
