| 宋羽霄,唐家琢,张志敏,等.肺腺癌细胞对RAW264.7巨噬细胞分化的影响[J].肿瘤学杂志,2022,28(6):452-458. |
| 肺腺癌细胞对RAW264.7巨噬细胞分化的影响 |
| Effect of Lung Adenocarcinoma Cells on the Polarization of Mouse Macrophage RAW264.7 Cells |
| 投稿时间：2021-12-05 |
| DOI：10.11735/j.issn.1671-170X.2022.06.B003 |
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| 中文关键词: 肺腺癌 RAW264.7 巨噬细胞 M2极化 补体C1q |
| 英文关键词:lung adenocarcinoma RAW264.7 macrophages M2 polarization complement C1q |
| 基金项目:希思科-BMS肿瘤免疫治疗研究基金(Y-BMS2019-003)；湖北省卫健委-武大人民联合科研课题 |
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| 中文摘要: |
| 摘 要：［目的］ 体外探究Lewis肺腺癌（Lewis lung carcinoma，LLC）细胞培养上清液对小鼠RAW264.7巨噬细胞极化的影响及其可能机制。［方法］ RAW264.7细胞按如下分组：空白对照组、IL-4处理组、LLC细胞培养上清液条件培养基（LLC-CM）培养组和TC-1细胞培养上清液条件培养基（TC-1-CM）培养组。各组培养48h后收集上清液和细胞，酶联免疫吸附测定（ELISA）检测空白对照组、IL-4组、LLC-CM组和TC-1-CM组巨噬细胞及LLC细胞和TC-1细胞培养上清液中补体C1q的含量，实时荧光定量PCR（qRT-PCR）检测巨噬细胞CD68、CD80、CD206、Arg-1、IL-10和IL-27 mRNA，Western blot检测CD68、CD80、CD206、Arg-1、JAK2、p-JAK2、STAT5和p-STAT5的表达。［结果］ 巨噬细胞经IL-4处理后，上清液中补体C1q含量较空白对照组升高（P<0.001）；与空白对照组及IL-4处理组相比，LLC-CM组巨噬细胞培养上清液中补体C1q含量明显升高（P均<0.001）。与空白对照组相比，LLC-CM组巨噬细胞CD80表达明显降低（P<0.001），CD206、Arg-1、IL-10和IL-27表达明显升高（P均<0.001），p-JAK2、p-STAT5（P均<0.001）含量降低。此外，LLC-CM组巨噬细胞p-JAK2、p-STAT5表达量低于IL-4处理组（P均<0.001）。［结论］ LLC细胞培养上清液可以通过抑制RAW264.7细胞JAK2/STAT5通路诱导其发生M2型极化并促进补体C1q高表达，高水平补体C1q可能参与促进RAW264.7细胞的M2型极化。 |
| 英文摘要: |
| Abstract：［Objective］ To explore the effect of Lewis lung adenocarcinoma(LLC) cells on the polarization of the mouse macrophages in vitro and its mechanism. ［Methods］ Mouse macrophage RAW264.7 cells were cultured with conditioned medium(control group) and the cultured cells were treated with IL-4(IL-4 group)， LLC cells culture supernatant(LLC-CM group) or TC-1 cells culture supernatant group(TC-1-CM group)， respectively. The content of complement C1q in the culture supernatant of 4 groups were detected with ELISA； the expression of CD68， CD80， CD206， Arg-1， IL-10， IL-27 mRNA was detected with real-time fluorescence quantitative RT-PCR(qRT-PCR)； and the expression of CD68， CD80， CD206， Arg-1， JAK2， p-JAK2， STAT5 and p-STAT5 proteins were detected by Western blot. ［Results］ After RAW264.7 cells were treated with IL-4， the content of complement C1q in the supernatant was higher than that in the blank control group(P<0.001). Compared with the blank control group and the IL-4 group， the content of complement C1q in LLC-CM group was significantly increased(P<0.001). Compared with the blank control group， the expressions of CD80 in LLC-CM group were significantly decreased(P<0.001)， while the expressions of CD206， Arg-1， IL-10 and IL-27 were significantly increased(all P<0.001)， and the levels of p-JAK2 and p-STAT5 were reduced(both P<0.001). Compared with the IL4 group， the levels of p-JAK2 and p-STAT5 in the LLC-CM group were decreased(both P<0.001). ［Conclusion］ LLC cell culture supernatant can induce M2 polarization in RAW264.7 cells by inhibiting the JAK2/STAT5 pathway and promoting the expression of complement C1q. |
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