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[目的]探索制备肺癌干细胞伴侣分子肿瘤抗原肽复合物的方法。[方法]采用肿瘤干细胞微球体培养分离法进行肺癌干细胞的培养与富集。选择鸦胆子油乳和125放射性碘粒子序贯处理肺癌干细胞,诱导伴侣分子抗原肽表达,同时选用普通肺癌细胞作为对照。采用交联—免疫沉淀法分离与鉴定伴侣分子抗原肽复合物制作成肺癌干细胞和普通肺癌细胞瘤苗。将肺癌干细胞与普通肺癌细胞进行荷瘤小鼠实验对比治疗效果。[结果]通过肺癌干细胞微球体培养分离法,成功培养出肺癌细胞微球体。结果显示鸦胆子油乳联合125放射性碘粒子可充分诱导肺癌细胞及肺癌干细胞中伴侣分子抗原肽的合成,且表达量与鸦胆子油乳的浓度成正比。肺癌细胞和肺癌干细胞中HSP70、HSP90表达与生理盐水比较,差异均有统计学意义(P<0.001)。两种细胞抑制肿瘤生长作用、延长小鼠生存期上亦均有统计学差异(P<0.05)。[结论]鸦胆子油乳联合125放射性碘粒子序贯处理和交联—免疫共沉淀法可充分诱导、分离与鉴定肺癌细胞及肺癌干细胞伴侣分子抗原肽的合成,肺癌干细胞较肺癌细胞更多地表达伴侣分子抗原肽,可提高肿瘤细胞的免疫原性,制作的瘤苗疗效更好,以期在防治肺癌和其复发上有较好的应用价值。
[Objective] To explore a method for preparing lung cancer stem cell partner tumor antigen peptide complex. [Method] Culture and enrichment of lung cancer stem cells using tumor stem cell microsphere culture separation method. Select Brucea javanica oil and 125 radioactive iodine particles sequential treatment of lung cancer stem cells, inducing chaperone antigen peptide expression, while the selection of ordinary lung cancer cells as a control. The cross-immunoprecipitation method was used to isolate and identify chaperone antigen peptide complexes to make lung cancer stem cells and ordinary lung cancer cell vaccine. Lung cancer stem cells and ordinary lung cancer cells were compared with the experimental treatment of tumor-bearing mice. [Result] Lung cancer cell microspheres were successfully cultivated by lung cancer stem cell microsphere culture separation method. The results showed that Brucea javanica oil combined 125 radioactive iodine particles could induce the synthesis of chaperone antigen peptide in lung cancer cells and lung cancer stem cells, and the expression level was proportional to the concentration of Brucea javanica oil emulsion. The expression of HSP70 and HSP90 in lung cancer cells and lung cancer stem cells were significantly different from those in normal saline (P <0.001). Two kinds of cells inhibit tumor growth and prolong the survival of mice also have statistical significance (P <0.05). [Conclusion] The combination of Brucea javanica oil emulsion and 125 radioactive iodine particle sequential treatment and cross-linking-co-immunoprecipitation method can induce, isolate and identify the synthesis of lung cancer cells and lung cancer stem cell chaperone antigen peptides. Lung cancer stem cells more than lung cancer cells Expression of peptide antigen peptide partners, can improve the immunogenicity of tumor cells, tumor vaccine produced better efficacy, with a view to prevention and treatment of lung cancer and its recurrence has a better value.