National Engineering Research Center for Nanomedicine, College of Life Science and Technology, Huazhong University of Science and Technology, 430074, Wuhan, China.
Hubei Bioinformatics & Molecular Imaging Key Laboratory, Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, 430074, Wuhan, China.
Oncogene. 2019 Oct;38(42):6818-6834. doi: 10.1038/s41388-019-0925-0. Epub 2019 Aug 12.
Tumor-repopulating cells (TRCs) are cancer stem cell (CSC)-like cells with highly tumorigenic and self-renewing abilities, which were selected from tumor cells in soft three-dimensional (3D) fibrin gels with unidentified mechanisms. Here we evaluated the transcriptome alteration during TRCs generation in 3D culture and revealed that a variety of molecules related with integrin/membrane and stemness were continuously altered by mechanical environment. Some key regulators such as MYC/STAT3/hsa-miR-199a-5p, were changed in the TRCs generation. They regulated membrane genes and the downstream mechanotransduction pathways such as Hippo/WNT/TGF-β/PI3K-AKT pathways, thus further affecting the expression of downstream cancer-related genes. By integrating networks for membrane proteins, the WNT pathway and cancer-related genes, we identified key molecules in the selection of TRCs, such as ATF4, SLC3A2, CCT3, and hsa-miR-199a-5p. Silencing ATF4 or CCT3 inhibited the selection and growth of TRCs whereas reduction of SLC3A2 or hsa-miR-199a-5p promoted TRCs growth. Further studies showed that CCT3 promoted cell proliferation and stemness in vitro, while its suppression inhibited TRCs-induced tumor formation. We also contemplated CCT3 as a stemness-related gene. Our findings provide insights in the mechanism of TRCs selection through transcriptome analysis.
肿瘤再生细胞（TRCs）是具有高致瘤性和自我更新能力的癌症干细胞（CSC）样细胞，它们是通过未明机制从软三维（3D）纤维蛋白凝胶中的肿瘤细胞中选择出来的。在这里，我们评估了 3D 培养中 TRC 生成过程中的转录组变化，并揭示了多种与整合素/膜和干性相关的分子被机械环境持续改变。一些关键调节剂，如 MYC/STAT3/hsa-miR-199a-5p，在 TRC 生成过程中发生了变化。它们调节膜基因和下游的机械转导途径，如 Hippo/WNT/TGF-β/PI3K-AKT 途径，从而进一步影响下游与癌症相关的基因表达。通过整合膜蛋白、WNT 途径和与癌症相关的基因网络，我们鉴定出了 TRC 选择中的关键分子，如 ATF4、SLC3A2、CCT3 和 hsa-miR-199a-5p。沉默 ATF4 或 CCT3 抑制了 TRC 的选择和生长，而减少 SLC3A2 或 hsa-miR-199a-5p 则促进了 TRC 的生长。进一步的研究表明，CCT3 促进了体外细胞增殖和干性，而其抑制则抑制了 TRC 诱导的肿瘤形成。我们还将 CCT3 视为与干性相关的基因。我们的研究结果通过转录组分析为 TRC 选择的机制提供了新的见解。
