| 王德佳,杨 波,高 星,刘惠苗,解冰川.IL-6在胶质母细胞瘤的表达及作用机制研究[J].现代生物医学进展英文版,2019,19(4):651-656. |
| IL-6在胶质母细胞瘤的表达及作用机制研究 |
| Expression and Potential Mechanisms of IL-6 in Malignant Glioma Cells |
| Received:April 08, 2018 Revised:April 30, 2018 |
| DOI：10.13241/j.cnki.pmb.2019.04.010 |
| 中文关键词: 白介素6 胶质母细胞瘤 p38MAPK MK2 |
| 英文关键词: IL-6 GBM p38MAPK MK2 |
| 基金项目:河北省卫生厅科研基金项目(20130580) |
| Author Name | Affiliation | E-mail | | WANG De-jia | Department of neurologyneurology, Central Hospital of Chengde, Chengde, Hebei, 067000, China | kloofishgood@163.com | | YANG Bo | Department of reproductive medicine, Central Hospital of Chengde, Chengde, Hebei, 067000, China | | | GAO Xing | Department of neurologyneurology, Central Hospital of Chengde, Chengde, Hebei, 067000, China | | | LIU Hui-miao | Department of neurologyneurology, First Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050031, China | | | XIE Bing-chuan | Department of neurologyneurology, First Hospital of Hebei Medical University, Shijiazhuang, Hebei, 050031, China | |
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| 中文摘要: |
| 摘要 目的：探讨白介素6(IL-6)在胶质母细胞瘤(GBM)中的表达，并探讨其高表达的作用机制，以期阐明GBM发生发展潜在分子机制。方法：采用免疫组化检测表皮生长因子变体3(EGFRvIII)阳性和阴性GBM组织IL-6的相对表达。以恶性胶质瘤细胞U87MG为研究对象，构建表达EGFRvIII的U87MG-EGFRvIII细胞，用IL-1β分别处理U87MG、U87MG-EGFRvIII细胞，ELISA检测IL-6分泌量。采用EGFR下游效应通路p38MAPK、MK2、MEK1/2、JNK抑制剂SB、sc-48、PD、SP预处理细胞1小时，IL-1β刺激细胞后，检测各组IL-6分泌量变化。将IL-1β处理后的U87MG-EGFRvIII细胞记为IL-1β组，以不做任何处理细胞记为Control组，用联合SB、sc-48处理的IL-1β细胞依次命名为IL-1β+SB和IL-1β+sc-48组，western blot检测p38MAPK-MK2通路蛋白和IL-6蛋白表达，qPCR检测IL-6 mRNA表达。结果：IL-6在EGFRvIII阳性GBM组织中普遍高表达，在EGFRvIII阴性GBM组织中普遍低表达。EGFRvIII可在未受IL-1β刺激的恶性胶质瘤细胞中上调IL-6基础分泌，也可在IL-1β刺激情况下进一步促进IL-6分泌。在U87MG细胞中，所有通路抑制剂对IL-6分泌均无影响；在U87MG-EGFRvIII细胞中p38 MAPK-MK2通路抑制剂SB和sc-48明显抑制IL-1β诱导的IL-6分泌，而MEK1/2、JNK抑制剂PD和SP则无明显影响。IL-1β能够诱导p38MAPK-MK2通路激活，诱导细胞内IL-6表达增加，联合SB、sc-48处理细胞后，p38MAPK-MK2通路活性降低，细胞内IL-6表达降低。结论：癌基因EGFRvIII能够上调恶性胶质瘤细胞中IL-6基础分泌，IL-1β可进一步刺激IL-6分泌，其机制可能与p38MAPK-MK2通路激活有关。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the mechanism of IL-6 overexpression in malignant glioma cells and clarify the potential molecular mechanism of GBM development. Methods: The relative level of IL-6 in EGFRvIII positive and negative GBM tissues were detected by immunohistochemistry. U87MG-EGFRvIII cells expressing EGFRvIII were constructed from U87MG malignant glioma cells. U87MG and U87MG-EGFRvIII cells were treated with IL-1β, and the secretion of IL-6 was detected by ELISA. The cells were pretreated with p38MAPK, MK2, MEK1/2, JNK inhibitor SB, sc-48, PD and SP for 1 hour, the changes of IL-6 secretion were measured after IL-1β treated. IL-1β-treated U87MG-EGFRvIII cells were recorded as IL-1β group, and U87MG-EGFRvIII cells were recorded as control group with no treatment. IL-1β cells treated with SB or sc-48 were named as IL-1β + SB and IL-1β + sc-48 group, respectively. The expression of p38MAPK-MK2 pathway protein and IL-6 protein were detected by western blot. The expression of IL-6 mR- NA was detected by qPCR. Results: IL-6 is generally high expressed in EGFRvIII-positive GBM tissues compared with that in EGFRvIII-negative GBM tissues. U87MG cells expressing the oncogene EGFRvIII were successfully constructed. The oncogene EGFRvIII can upregulate IL-6 basal secretion in glioma cells, and further promote IL-6 secreting under the stimulation of IL-1β. In U87MG cells, all inhibitors had almost no effect on IL-6 secretion; p38 MAPK-MK2 pathway inhibitors SB, sc-48 significantly inhibited IL-1β-induced IL-6 secretion in U87MG-EGFRvIII cells while MEK1/2, JNK inhibitor PD, SP had no significant effect. IL-1β-treated cells could in- duce the activation of p38MAPK-MK2 pathway and induce the expression of intracellular IL-6. The activity of p38MAPK-MK2 pathway and the expression of IL-6 were inhibited after IL-1β combinated with SB and sc-48. Conclusion: The oncogene EGFRvIII can upregulate IL-6 basal secre- tion in glioma cells with no IL-1β treatment, and further promote the secretion of IL-6 under the stimulation of IL-1β, which may be related to the activation of p38MAPK-MK2 pathway. Our research revealing that glioma cells may form inflammatory microenvironment through endogenous (EGFRvIII) and exogenous (IL-1β) factors, and provides the possibility for further improvement of brain-permeable and anti-inflammatory inhibitors targeting p38 MAPK, MK2 to improve the progression of aggressive gliomas. |
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