| 王 浩,黄常新,朱 影.序贯处理交联—免疫共沉淀法制备肺癌干细胞伴侣分子抗原肽瘤苗的研究[J].肿瘤学杂志,2015,21(12):982-987. |
| 序贯处理交联—免疫共沉淀法制备肺癌干细胞伴侣分子抗原肽瘤苗的研究 |
| Study of Lung Cancer Stem Cells Partner Molecular Antigen Peptide Tumor Vaccine Through the Method of Sequential Processing and Crosslinking-immunprecipitation |
| 投稿时间：2015-07-15 |
| DOI：10.11735/j.issn.1671-170X.2015.12.B008 |
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| 中文关键词: 肺癌干细胞 抗癌瘤苗 碘 鸦胆子油乳 交联—免疫共沉淀 |
| 英文关键词:lung cancer stem cells anti-cancer vaccine iodine Brucea javanica oil emulsion crosslinking-immunprecipitation |
| 基金项目:浙江省科技厅公益技术研究社会发展项目（2011C33030） |
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| 中文摘要: |
| 摘 要：［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。 |
| 英文摘要: |
| Abstract：［Purpose］ To explore the prepare method of lung cancer stem cells partner molecular tumor antigen peptide.［Methods］ Cancer stem cells were cultured and enriched by the conventional microsphere separation process. Lung cancer stem cells were sequential processed with Brucea oil emulsion and 125I seed，in order to induce express of chaperone antigenic peptide. Normal lung cancer cells were also selected as a control at the same time. Isolation and identification of chaperone peptide complex through crosslinking-immunprecipitation made into lung cancer stem cells and normal lung tumor vaccines. The efficacy of treatment of lung cancer stem cells and lung cancer cells through tumor-burdened experiment on mice was compared.［Results］ Lung cancer cells microspheres were successfully cultured by lung cancer stem cell microspheres cultivate separation. Experimental results showed that the United of Brucea oil emulsion and particles of radioactive iodine 125 could induce cancer cells and cancer stem cells chaperone antigenic peptides synthesis，and the expression was proportional to the concentration of Brucea oil emulsion. The expression of HSP70，HSP90 was different in the saline，lung cancer cells and cancer stem cells groups(P<0.001). The effect on inhibiting tumor growth was also significantly different (P<0.05). The effect of prolong the mice survival was also significantly different(P<0.05).［Conclusion］ Brucea oil emulsion combined with radioactive iodine particles can fully induce the synthesis of 125I lung cancer cells and cancer stem cells chaperone antigenic peptides and isolation and identification of lung cancer cells and cancer stem cells. Lung cancer stem cells express more chaperone antigenic peptides than cancer cells，also can improve the immunogenicity of the tumor cells. Tumor vaccines could make stronger effect on inhibiting tumor growth and prolonging the survival period of mice. |
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