Abstract
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.
Highlights
Cancer stem cells (CSCs) are a small subpopulation of cancer cells identified in both hematologic malignancies and solid tumor types with high tumorigenicity and high capabilities of self-renewal [1]
These findings suggest that HeLa Tumor-repopulating cells (TRCs) in 3D soft fibrin gels are unique in promoting multicellular tumor spheroid formation and growth with enhanced self-renewing capacities
The silence of SLC3A2 increased the colony number and size (Fig. 4g). These results suggested that ATF4 and CCT3 promoted the selection of HeLa TRCs, while SLC3A2 suppressed it
Summary
Cancer stem cells (CSCs) are a small subpopulation of cancer cells identified in both hematologic malignancies and solid tumor types with high tumorigenicity and high capabilities of self-renewal [1]. The process of TRCs selection/transformation is a very good model to study the origin, feature and mechanism of CSCs. a comprehensive study on expression and regulation of the origin of TRCs may give us insights into the acquisition of cancer stemness and potential targets for prevention and treatment of cancer. The transcriptome profiling and regulatory networks of the TRCs selection process could provide a comprehensive landscape to the underlying molecular and regulatory mechanisms and provide new insights to cancer stemness. To better understand the TRCs selection and transformation, we sequenced the transcriptome and miRNA for samples that collected during the soft fibrin gels selection process, followed by systematical bioinformatics analysis on expression and regulation. It showed a comprehensive view of the gene expression and regulation for TRCs selection and provided potential targets for cancer therapy
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
