Jiangsu Center for Drug Screening, China Pharmaceutical University, Nanjing, 210009, China.
State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, People's Republic of China.
Clin Transl Oncol. 2020 Sep;22(9):1591-1602. doi: 10.1007/s12094-020-02303-z. Epub 2020 Feb 13.
Tumor metastasis is a terrifying characteristic of cancer. Numerous studies have been conducted to overcome metastasis by targeting tumor microenvironment (TME). However, due to complexity of tumor microenvironment, it remained difficult for accurate targeting. Dwarf-lillytruf tuber monomer-13 (DT-13) possess good potential against TME.
As TME is supportive for tumor metastasis, alternatively it is a challenging for therapeutic intervention. In our present study, we explored molecular mechanism through which TME induced cell migration and how DT-13 interferes in this mechanism.
We used a novel model of co-culture system which is eventually developed in our lab. Tumor cells were co-cultured with hypoxia induced cancer-associated fibroblasts (CAF) or with chemically induced cancer-associated adipocytes (CAA). The effect of hypoxia in conditioned medium for CAF was assessed through expression of α-SMA and HIF by western blotting while oil red staining was done to assess the successful chemical induction for adipocytes (CAA), the effect of TME through conditioned medium on cell migration was analyzed by trans-well cell migration, and cell motility (wound healing) analyses. The expression changes in cellular proteins were assessed through western blotting and immunofluorescent studies.
Our results showed that tumor microenvironment has a direct role in promoting breast cancer cell migration by stromal cells; moreover, we found that DT-13 restricts this TME regulated cell migration via targeting stromal cells in vitro. Additionally we also found that DT-13 targets NMII-A for its effect on breast cancer cell migration for the regulation of stromal cells in TME.
肿瘤转移是癌症的一个可怕特征。为了克服肿瘤微环境（TME）的影响，已经进行了许多研究。然而，由于肿瘤微环境的复杂性，对其进行准确靶向仍然很困难。矮百合菝葜单体-13（DT-13）对 TME 具有很好的靶向潜力。
由于 TME 支持肿瘤转移，因此对其进行治疗干预具有挑战性。在本研究中，我们探讨了 TME 诱导细胞迁移的分子机制，以及 DT-13 如何干扰这一机制。
我们使用了一种新型共培养系统模型，该模型最终由我们实验室开发。将肿瘤细胞与缺氧诱导的癌症相关成纤维细胞（CAF）或化学诱导的癌症相关脂肪细胞（CAA）共培养。通过 Western blot 检测 CAF 条件培养基中 α-SMA 和 HIF 的表达，评估缺氧对 CAF 的影响；通过 Transwell 细胞迁移和细胞迁移（划痕愈合）分析，评估 TME 通过条件培养基对细胞迁移的影响；通过 Western blot 和免疫荧光研究评估细胞蛋白表达变化。
结果表明，肿瘤微环境通过基质细胞直接促进乳腺癌细胞迁移；此外，研究发现 DT-13 通过靶向体外 TME 调节的细胞迁移中的基质细胞来限制这种 TME 调节的细胞迁移。此外，研究还发现 DT-13 通过靶向 NMII-A 来调节 TME 中的基质细胞，从而影响乳腺癌细胞迁移。
