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Distinguishing high-metastasis-potential circulating tumor cells through fluidic shear stress in a bloodstream-like microfluidic circulatory system

Oncogene volume 43, pages 2295–2306 (2024)Cite this article

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Abstract

Circulating tumor cells (CTCs) play a critical role as initiators in tumor metastasis, which unlocks an irreversible process of cancer progression. Regarding the fluid environment of intravascular CTCs, a comprehensive understanding of the impact of hemodynamic shear stress on CTCs is of profound significance but remains vague. Here, we report a microfluidic circulatory system that can emulate the CTC microenvironment to research the responses of typical liver cancer cells to varying levels of fluid shear stress (FSS). We observe that HepG2 cells surviving FSS exhibit a marked overexpression of TLR4 and TPPP3, which are shown to be associated with the colony formation, migration, and anti-apoptosis abilities of HepG2. Furthermore, overexpression of these two genes in another liver cancer cell line with normally low TLR4 and TPPP3 expression, SK-Hep-1 cells, by lentivirus-mediated transfection also confirms the critical role of TLR4 and TPPP3 in improving colony formation, migration, and survival capability under a fluid environment. Interestingly, in vivo experiments show SK-Hep-1 cells, overexpressed with these genes, have enhanced metastatic potential to the liver and lungs in mouse models via tail vein injection. Mechanistically, TLR4 and TPPP3 upregulated by FSS may increase FSS-mediated cell survival and metastasis through the p53-Bax signaling pathway. Moreover, elevated levels of these genes correlate with poorer overall survival in liver cancer patients, suggesting that our findings could offer new therapeutic strategies for early cancer diagnosis and targeted treatment development.

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Fig. 1: Responses of HepG2 cells to FSS.
Fig. 2: Exploration of differentially expressed genes in FSS-treated HepG2 cells by RNA-seq and RT-qPCR.
Fig. 3: Effect of downregulating TLR4 or TPPP3 on the survival in circulation, colony formation, migration, and anti-anoikis of HepG2 cells.
Fig. 4: Responses of SK-Hep-1 cells to FSS.
Fig. 5: Responses of TLR4 or TPPP3-overexpressing SK-Hep-1 cells in vitro and in vivo.
Fig. 6: Clinical significance of TPPP3 and TLR4 in liver cancer.

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All data supporting the findings of this work are available from the main text article and its Supplementary Information files or from the corresponding author on reasonable request.

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Acknowledgements

This work was supported by grants from Hetao Shenzhen-Hong Kong Science and Technology Innovation Cooperation Zone Shenzhen Park Project (HZQB-KCZYZ-2021017), National Key Research and Development Program of China (2018YFA0901104), and City University of Hong Kong (project #9680217 and # 9678223). The authors thank Wang Peng, Zesheng Wang, Tongxu Si, and NG Ka Ki for their assistance in experiments.

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  1. These authors contributed equally: Wenxiu Li, Zhengjun Guo, Zhihang Zhou.

Authors and Affiliations

  1. Department of Precision Diagnostic and Therapeutic Technology, City University of Hong Kong Shenzhen Futian Research Institute, Shenzhen, 518057, China

    Wenxiu Li, Zhengjun Guo, Zhengdong Zhou, Huimin He, Jiayu Sun, Xiaoyu Zhou, Y. Rebecca Chin, Liang Zhang & Mengsu Yang

  2. Department of Biomedical Sciences, and Tung Biomedical Sciences Centre, City University of Hong Kong, 83 Tat Chee Avenue, Kowloon, Hong Kong SAR, 999077, China

    Wenxiu Li, Zhengdong Zhou, Huimin He, Jiayu Sun, Xiaoyu Zhou, Y. Rebecca Chin, Liang Zhang & Mengsu Yang

  3. Department of Cancer Center, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

    Zhengjun Guo

  4. Department of Gastroenterology, The Second Affiliated Hospital of Chongqing Medical University, Chongqing, 400010, China

    Zhihang Zhou

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Contributions

Wenxiu Li is responsible for conducting the experiments, collecting data, plotting figures, and writing the manuscript. Zhengjun Guo and Zhihang Zhou are responsible for conducting animal experiments and reviewing the manuscript. Zhengdong Zhou and Huimin He contributed to result discussion and writing the manuscript. Jiayu Sun, Xiaoyu Zhou, Y. Rebecca Chin, Liang Zhang provided feedback on the manuscript. Mengsu Yang is responsible for designing, supervising, and writing this study.

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Correspondence to Mengsu Yang.

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All animal experiments were conducted according to the animal protocol approved by the Animal Care and Ethics Committee of City University of Hong Kong (A-0644). All experiments about human samples were approved by the Ethical Committee of the Second Affiliated Hospital of Chongqing Medical University (2022)80) and carried out in accordance with the Declaration of Helsinki. Written informed consent was obtained from all participants.

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Li, W., Guo, Z., Zhou, Z. et al. Distinguishing high-metastasis-potential circulating tumor cells through fluidic shear stress in a bloodstream-like microfluidic circulatory system. Oncogene 43, 2295–2306 (2024). https://doi.org/10.1038/s41388-024-03075-4

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