Characterization of the inflammatory response during Ehrlich ascitic tumor development
Characterization of the inflammatory response during Ehrlich ascitic tumor development
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DOI：
10.1016/j.vascn.2014.09.001
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发表时间：
2015-01-01
影响因子：
1.9
通讯作者：
Neves, Josiane S.
中科院分区：
文献类型：
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作者：
Fernandes, Patricia Dias;Guerra, Fabiana S.;Neves, Josiane S.
前言：埃利希瘤是一种具有侵袭性的乳腺腺癌。小鼠腹腔接种埃利希瘤，瘤体呈腹水状生长。由于炎症调节肿瘤进展，我们进一步研究了EAT生长过程中的炎症反应。研究方法：Balb/C小鼠腹腔内接种5 × 10（5）个埃利希细胞，每2天后采集血样用于血红蛋白、血细胞比容、血小板和白细胞计数。收集腹水用于蛋白浓度和细胞计数。采用流式细胞仪（FACS）、酶联免疫吸附试验（ELISA）检测前列腺素E2（PGE 2）和细胞因子，硝酸盐转化法检测一氧化氮（NO），免疫印迹法检测环氧合酶1（COX 1）、COX 2和诱导型一氧化氮合酶（iNOS）。结果：腹腔接种EAT后，腹水体积和蛋白浓度迅速增加。腹水中的细胞数量保持稳定，直到第8天（滞后期），随后急剧增加。随着肿瘤的进展，血液白细胞增加，红细胞减少。表型分析表明，在滞后期，F4/80（+）细胞的百分比保持与对照水平相似，约7%的该群体也对GR 1标记物呈阳性。这些双阳性细胞（可能是炎性单核细胞）在第6天显著增加。F4/80-GR 1（+）细胞（可能是中性粒细胞）的百分比较低，并且在肿瘤进展期间没有显著变化。在分析的时间点未检测到CD 4（+）和CD 8（+）细胞。iNOS和COX 1表达在第2天后增加，在第10天达到峰值水平。COX 2酶的表达没有随时间发生显著变化。观察到PGE 2和NO水平持续升高。IL-10和MCP-1在第14天达到峰值，IL-1 β逐渐增加直至第10天。IFN-γ水平低，直到第10天，之后逐渐增加。讨论内容：这些数据扩展了埃利希腹水瘤生长过程中炎症反应的表征，进一步验证了其作为抗肿瘤药物筛选的有用模型。(C)2014爱思唯尔公司All rights reserved.
Introduction: Ehrlich tumor is a mammary adenocarcinoma with aggressive behavior. Inoculated in mice peritoneal cavity, the Ehrlich tumor grows in ascitic form (EAT). Since inflammation modulates tumor progression we further investigated the inflammatory response during EAT growth. Methods: Balb/C mice were intraperitoneal inoculated with 5 x 10(5) Ehrlich cells and after every 2 days, blood samples were collected for hemoglobin, hematocrit, platelets and leukocytes counts. The ascitic fluid was collected for protein concentration and cell count. Phenotype analysis of the peritoneal cells was made by FACS, prostaglandin E2 (PGE2) and cytokines by ELISA, nitric oxide (NO) by nitrate conversion protocol, and cyclooxygenase-1 (COX1), COX2 and inducible nitric oxide synthase (iNOS) by immunoblotting. Results: Following EAT inoculation into the peritoneal cavity there was a rapid increase in ascitis volume and protein concentration. The cell number in ascitis remained stable until day 8 (lag phase) followed by a sharp increase. As tumor progressed, blood leukocytes increased and erythrocyte decreased. Phenotypic analysis showed that during the lag phase the percentage of F4/80(+) cells remained similar to control levels and around 7% of this population was also positive for the GR1 marker. These double-positive cells (probably inflammatory monocytes) markedly increased at day 6. The percentage of F4/80-GR1(+) cells (probably neutrophils) was low and did not significantly vary during tumor progression. CD4(+) and CD8(+) cells were not detected in the time points analyzed. iNOS and COX1 expression increased after day 2 reaching peak levels on day 10. COX2 enzyme expression did not change significantly over time. Sustained increase in PGE2 and NO levels was observed. IL-10 and MCP-1 peaked at day 14 and IL-1 beta increased progressively till day 10. IFN-gamma levels were low till day 10, increasing progressively after that. Discussion: These data extended the characterization of the inflammatory response during Ehrlich ascitis tumor growth, further validating it as a useful model for antitumor drugs screening. (C) 2014 Elsevier Inc. All rights reserved.