| 摘要: |
| ［摘要］ 目的 探讨胶质母细胞瘤（GBM）中癌-睾丸抗原OY-TES-1的表达对M2型巨噬细胞极化的影响。方法 体外培养人GBM细胞U251与人外周血单核细胞THP-1，构建稳定下调OY-TES-1的U251稳转株（U251-SH-OY-TES-1组），将转染无关序列的U251作为对照（U251-SH-NC组）。48 h后取两组上清液与THP-1细胞共培养，加入U251-SH-NC组上清液的THP-1细胞为SH-NC组，加入U251-SH-OY-TES-1组上清液的THP-1细胞为SH-OY-TES-1组。通过实时荧光定量聚合酶链反应（RT-qPCR）和蛋白免疫印迹法分别检测两组OY-TES-1 mRNA和蛋白相对表达量，确定转染效率。通过酶联免疫吸附试验（ELISA）检测培养液中人白细胞介素-10（IL-10）、人转化生长因子-β（TGF-β）表达水平。通过流式细胞术检测CD206表达水平。结果 RT-qPCR与蛋白免疫印迹法检测结果显示，U251-SH-OY-TES-1组OY-TES-1 mRNA和蛋白的相对表达量低于U251-SH-NC组，差异有统计学意义（P<0.05）。ELISA结果显示，两组培养液中TGF-β、IL-10水平随培养时间的增加呈上升趋势。第0天、第2天SH-OY-TES-1组培养液中TGF-β水平显著低于SH-NC组（P<0.05），IL-10水平比较差异无统计学意义（P>0.05）。第4天SH-OY-TES-1组培养液中TGF-β、IL-10水平显著低于SH-NC组（P<0.05）。流式细胞术结果显示，与SH-NC组比较，SH-OY-TES-1组CD206水平下降，差异有统计学意义（P<0.05）。结论 GBM中OY-TES-1高表达能够促进M2型巨噬细胞的极化。 |
| 关键词: 胶质母细胞瘤 OY-TES-1 M2型巨噬细胞 极化 |
| DOI：10.3969/j.issn.1674-3806.2024.08.07 |
| 分类号: |
| 基金项目:国家自然科学基金地区科学基金项目（编号：81960453）；广西自然科学基金项目（编号：2023GXNSFAA026120） |
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| Effect of cancer-testis antigen OY-TES-1 expression on the polarization of M2 macrophages in glioblastoma |
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ZHAO Zhenkai1, LIANG Guo1, LI Feng1, NONG Weixia1,2, ZHANG Qingmei1,2, LUO Bin1, XIE Xiaoxun1,2
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1.Teaching and Research Section of Histology and Embryology, Guangxi Medical University, Nanning 530021, China; 2.Key Laboratory of Basic Research on Regional Diseases in Guangxi Colleges and Universities, Nanning 530021, China
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| Abstract: |
| ［Abstract］ Objective To explore the effect of cancer-testis antigen OY-TES-1 expression on the polarization of M2 macrophages in glioblastoma(GBM). Methods Human GBM cell U251 and human peripheral blood mononuclear cell THP-1 were cultured in vitro to construct a stable transfection cell line U251 that stably down-regulated OY-TES-1(U251-SH-OY-TES-1 group), and U251 cells transfected with unrelated sequences were used as the controls(U251-SH-NC group). Forty-eight hours later, the supernatant of the two groups was co-cultured with THP-1 cells. THP-1 cells to which the supernatant of the U251-SH-NC group was added were taken as the SH-NC group, and THP-1 cells to which the supernatant of the U251-SH-OY-TES-1 group was added were taken as the SH-OY-TES-1 group. The relative expression levels of OY-TES-1 mRNA and protein in the two groups were detected by using real-time fluorescence quantitative polymerase chain reaction(RT-qPCR) and Western blotting, respectively, and the transfection efficiency was determined. The expression levels of human interleukin-10(IL-10) and human transforming growth factor-β(TGF-β) in the culture solution were detected by enzyme-linked immunosorbent assay(ELISA). The expression level of CD206 was detected by flow cytometry. Results The detection results of RT-qPCR and Western blotting method showed that the relative expressions of OY-TES-1 mRNA and protein in the U251-SH-OY-TES-1 group were lower than those in the U251-SH-NC group, and the differences were statistically significant(P<0.05). The results of ELISA showed that the levels of TGF-β and IL-10 in the culture solution of the two groups increased with the increase of culture time. On day 0 and day 2, the levels of TGF-β in the culture solution of the SH-OY-TES-1 group were significantly lower than those of the SH-NC group(P<0.05), and there were no significant differences in the levels of IL-10 on day 0 and day 2 between the two groups(P>0.05). On day 4, the levels of TGF-β and IL-10 in the culture solution of the SH-OY-TES-1 group were significantly lower than those of the SH-NC group(P<0.05). The results of flow cytometry showed that compared with that in the SH-NC group, the level of CD206 in the SH-OY-TES-1 group decreased, and the difference was statistically significant(P<0.05). Conclusion The high expression of OY-TES-1 in GBM can promote the polarization of M2 macrophages. |
| Key words: Glioblastoma(GBM) OY-TES-1 M2 macrophage Polarization |
