顺铂对肿瘤间充质干细胞中IL-6表达的调控及p38MAPK通路参与机制

doi:10.6040/j.issn.1671-7554.0.2018.1233

摘要/Abstract

摘要： 目的探讨顺铂(CDDP)对肿瘤间充质干细胞(MSCs)中白细胞介素-6(IL-6)表达的调控作用与参与机制。方法采用CDDP分别处理小鼠Lewis肺癌(LLC)细胞和MSCs,聚合酶链反应阵列(PCR Array)检测、筛选其IL-6表达改变,并通过实时荧光定量聚合酶链反应(qRT-PCR)、酶联免疫吸附实验(ELISA)等方法进一步验证。Western blotting检测CDDP处理后MSCs中p38MAPK通路的表达改变;并通过该通路特异性抑制剂抑制其激活,ELISA检测IL-6的表达。结果 CDDP处理LLC细胞后,IL-6的表达未见有统计学意义的改变(P>0.05)。PCR Array结果显示,CDDP处理MCSs 24 h后,各白细胞介素因子有不同程度的表达改变,其中IL-6表达上升最为显著;进一步研究证实,CDDP作用于MCSs 3、6和24 h后,IL-6 mRNA和蛋白表达均升高,差异具有统计学意义(IL-6 mRNA:P=0.218;P=0.007;P<0.001;蛋白表达:P=0.001;P<0.001;P<0.001)。CDDP处理后MSCs中p38和pp38的表达均明显增强,且pp38/p38比值明显升高;采用SB203580抑制p38MAPK通路后,CDDP所致的IL-6上调表达被部分逆转。结论 CDDP对LLC细胞中IL-6的表达无统计学意义影响,但可显著上调MSCs中IL-6的表达。CDDP处理后,MSCs中p38MAPK通路表达上调,抑制该通路后可下调IL-6的表达,提示CDDP可能通过p38MAPK通路调控IL-6在MSCs的表达。

关键词: 肺癌, 顺铂, 白细胞介素6, 间充质干细胞, p38MAPK通路

Abstract: Objective To investigate the effect of cisplatin(CDDP)on interleukin(IL)-6 expression in tumor mesenchymal stem cells(MSCs)and explore its possible mechanisms. Methods Lewis lung carcinoma(LLC)cells or MSCs were treated with CDDP, and their IL gene expression was detected by polymerase chain reaction array(PCR Array)and further verified by quantitative real-time polymerase chain reaction(qRT-PCR)and enzyme-linked immunosorbent assay(ELISA). After CDDP treatment, the activation of p38MAPK pathway in MSCs was analyzed by Western blotting. CDDP activated p38MAPK pathway was inhibited by the specific inhibitor, and IL-6 expression in MSCs was measured by ELISA. Results CDDP treatment showed no significant effect on the expression of IL-6 in LLC cells(P>0.05). However, IL gene expression was changed in MSCs treated with CDDP for 24 h, and IL-6 expression was increased most significantly. Further studies confirmed the increase of IL-6 mRNA and protein in MSCs after CDDP treatment for 3 h, 6 h and 24 h(P<0.05). Higher expressions of p38 and pp38 were detected in MSCs after CDDP treatment. Moreover, the ratio of pp38/p38 was significantly increased. After the p38MAPK pathway was inhibited by SB203580, the up-regulation of IL-6 caused by CDDP was partially reversed. Conclusion CDDP has no significant effect on IL-6 expression in LLC cells, but it significantly up-regulates the expression of IL-6 in MSCs. CDDP treatment activates the p38MAPK pathway in MSCs, and inhibiting this pathway resultes in down-regulation of IL-6. These results indicate that CDDP might regulate the expression of IL-6 in MSCs via the p38MAPK pathway.

Key words: Lung cancer, Cisplatin, Interleukin-6, Mesenchymal stem cells, p38MAPK pathway

中图分类号:

R734.2

张志慧,刘延国,肖丰启,马道新,王秀问. 顺铂对肿瘤间充质干细胞中IL-6表达的调控及p38MAPK通路参与机制[J]. 山东大学学报 (医学版), 2019, 57(3): 1-6.

ZHANG Zhihui, LIU Yanguo, XIAO Fengqi, MA Daoxin, WANG Xiuwen. Effect of cisplatin on IL-6 expression in tumor mesenchymal stem cells and underlying mechanism of p38MAPK pathway[J]. Journal of Shandong University (Health Sciences), 2019, 57(3): 1-6.

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