Bian Huan, Zhang Shuai, Wu Huanhuan, Wang Yixiang
Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology, 22 Zhongguancun Avenue South, Haidian District, Beijing, 100081, China.
Tumour Biol. 2015 Jun;36(6):4467-77. doi: 10.1007/s13277-015-3088-9. Epub 2015 Mar 10.
The influence of tumor surrounding microenvironment is often neglected when immunohistochemistry is performed to investigate tumor properties and search biomarkers of cancer. This study was designed to evaluate whether the influence of tumor microenvironment on biological features of tumor cells should be taken into account for interpretation of the immunohistochemistry data of tumor specimens. In this study, we showed an example by using three tumor cell lines (HeLa, WSU-HN6, and Tca83) to establish tumor-caused bone destruction models in nude mice and then to investigate the influence of bone marrow microenvironment (BMM) on biological features of tumor cells. Immunohistochemistry results showed that, compared with tumor cells located outside of BMM, tumor cells located inside of BMM presented huge differences in the expression of inflammation-related proteins including tumor necrosis factor-α (TNF-α), TNF receptor-associated factor protein-6 (TRAF-6), phosphorylated-NF-κB p65 (p-p65), interleukin (IL)-6 and IL-11, matrix metalloproteinases including MMP-1, MMP-2, MMP-9, and MMP-13; and osteogenesis-related proteins including runt-related transcription factor 2 (RUNX2), bone sialoprotein (BSP), and osteocalcin (OCN) in all the models. However, when we compared the cell line pair derived from different sites (outside and inside of BMM, respectively) of the same HeLa tumor sample by real-time PCR, Western blot, and immunocytochemistry, the differences aforementioned in tumor tissues were not found. In addition, we verified that normal human bone marrow could not cause the above changes detected in vivo. Our results suggested that tumor-modified microenvironment could give the new biological features of the invaded tumor cells. Therefore, we should consider the influence of the surrounding microenvironment on tumor cells when we analyze tumor properties using immunohistochemistry.
在通过免疫组织化学研究肿瘤特性并寻找癌症生物标志物时，肿瘤周围微环境的影响常常被忽视。本研究旨在评估在解释肿瘤标本的免疫组织化学数据时，是否应考虑肿瘤微环境对肿瘤细胞生物学特性的影响。在本研究中，我们通过使用三种肿瘤细胞系（HeLa、WSU-HN6和Tca83）在裸鼠中建立肿瘤导致的骨破坏模型，进而研究骨髓微环境（BMM）对肿瘤细胞生物学特性的影响，展示了一个实例。免疫组织化学结果显示，与位于BMM外的肿瘤细胞相比，位于BMM内的肿瘤细胞在所有模型中，炎症相关蛋白（包括肿瘤坏死因子-α（TNF-α）、TNF受体相关因子蛋白-6（TRAF-6）、磷酸化核因子κB p65（p-p65）、白细胞介素（IL）-6和IL-11）、基质金属蛋白酶（包括MMP-1、MMP-2、MMP-9和MMP-13）以及成骨相关蛋白（包括 runt相关转录因子2（RUNX₂）、骨唾液蛋白（BSP）和骨钙素（OCN））的表达存在巨大差异。然而，当我们通过实时PCR、蛋白质印迹和免疫细胞化学比较来自同一HeLa肿瘤样本不同部位（分别为BMM外和BMM内）的细胞系对时，未发现肿瘤组织中上述差异。此外，我们证实正常人骨髓不会引起体内检测到的上述变化。我们的结果表明，肿瘤修饰的微环境可赋予侵袭性肿瘤细胞新的生物学特性。因此，在使用免疫组织化学分析肿瘤特性时应考虑周围微环境对肿瘤细胞的影响。
