反义人IL-10和反义病毒性IL-10抗鼻咽癌细胞成瘤效应的研究

The study of anti-tumor genecity effect of human IL-10 antisense and viral IL-10 antisense on nasopharyngeal carcinoma cells

目的:探讨反义人IL-10和反义病毒性IL-10抑制鼻咽癌细胞成瘤的效果。方法:扩增及克隆hIL-10 cDNA和 vIL-10基因,构建反义hIL-10和反义vIL-10双价真核表达载体pcDNA3/AS hIL-10+AS vIL-10,转染鼻咽癌细胞株SUNE。将转染阳性的SUNE细胞克隆和未转染的SUNE细胞接种SCID小鼠和经健康人外周血白细胞免疫重建的SCID小鼠（hu-PBL-SCID鼠）,观察成瘤效应。结果:ELISA法测得未转染SUNE细胞培养上清液中IL-10和vIL-10的总含量为242±23pg/ml,而转染阳性的SUNE细胞培养上清液中hIL-10和vIL-10的总含量仅为22±6pg/ml。在SCID鼠体内,vIL-10和hIL-10的表达与否,并不影响鼻咽癌细胞的成瘤活性,但在hu-PBL-SCID鼠体内,抑制vIL-10和hIL-10的表达则可明显抑制鼻咽癌细胞的成瘤作用。结论:反义人IL-10和反义病毒性IL-10可显著抑制鼻咽癌细胞在血hu-PBL-SCID鼠体内的成瘤作用,鼻咽癌组织表达vIL-10和hIL-10的确与鼻咽癌免疫耐受有关。

Objective: To study the effectiveness of anti-tumorgenecity of human IL-10 antisense and viral IL-10 antisense on nasopharyn-geal carcinoma(NPC) cells.Methods:Human IL-10 cDNA and viral IL-10 gene had been amplifed and cloned. A two-valent eukaryotic expression vector for antisense nucleic acids of human IL-10 cDNA and viral IL-10 gene had been constructed and designated as pcDNA3/AS hIL-10 + AS vIL-10.The recombinant vector was transfected into a NPC-clerived cell strain-SUNE cells. After that, the transfected and nontransfected SUNE cells were inoculated i.d. in the right flank of SCID mice and hu-PBL-SCID mice to evaluate their tumorgenecity changes.Results:The results of ELISA analysis showed that the concentration of hIL-10 and vIL-10 was 242 ?3 pg'ml-1 in supernatant of the nontransfected SUNE cells, and onry 22 ? pg ?ml-1 of hIL-10 and vIL-10 contained in supernatant of the transfected SUNE cells. After inoculation to SCID mice, the transfected and nontransfected SUNE cell showed the same growth manner. But inoculating to hu-PBL-SCID mice, the transfected SUNE cells grew slowerly than the nontransfected SUNE cell. With statistical analysis, the size and mean weight of the tumors from the hu-PBL-SCID mice inoculated with transfected SUNE cells were significantly lower in contrast to that from the hu-PBL-SCID mice inoculated with nontransfected SUNE cells( P < 0.01). Conclusion: The expression blocking of IL-10 through their antisense nucleic acids can significantly inhibit the tumorgenecity of NPC cells in hu-PBL-SCID mice model but not in SCID mice hIL-10 and vIL-10 expressed in NPC tissues can certainly offer a favourable cytokine milieu for the tumor cells escaping from the attack by immune competent cells.