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SHMT2 promotes cell viability and inhibits ROS-dependent, mitochondrial-mediated apoptosis via the intrinsic signaling pathway in bladder cancer cells

Cancer Gene Therapy volume 29, pages 1514–1527 (2022)Cite this article

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Abstract

Mitochondrial serine hydroxymethyltransferase (SHMT2) catalyzes the conversion of serine to glycine and concomitantly produces one-carbon units to support cell growth and is upregulated in various cancer cells. SHMT2 knockdown triggers cell apoptosis; however, the detailed mechanism of apoptosis induced by SHMT2 inactivation remains unknown. Here, we demonstrate that SHMT2 supports the proliferation of bladder cancer (BC) cells by maintaining redox homeostasis. SHMT2 knockout decreased the pools of purine and one-carbon units and delayed cell cycle progression in a manner that was rescued by formate, demonstrating that SHMT2-mediated one-carbon units are essential for BC cell proliferation. SHMT2 deficiency promoted the accumulation of intracellular reactive oxygen species (ROS) by decreasing the NADH/NAD+, NADPH/NADP+, and GSH/GSSG ratios, leading to a loss in mitochondrial membrane potential, release of cytochrome c, translocation of Bcl-2 family protein and activation of caspase-3. Notably, blocking ROS production with the one-carbon donor formate and the ROS scavenger N-acetyl-cysteine (NAC) effectively rescued SHMT2 deficiency-induced cell apoptosis via the intrinsic signaling pathway. Treatment with the SHMT inhibitor SHIN1 resulted in a significant inhibitory effect on cell proliferation and induced cell apoptosis. Formate and NAC rescued SHIN1-induced cell apoptosis. Our findings reveal an important mechanism by which the loss of SHMT2 triggers ROS-dependent, mitochondrial-mediated apoptosis, which gives insight into the link between serine metabolism and cell apoptosis and provides a promising target for BC treatment and drug discovery.

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Fig. 1: SHMT2 expression is upregulated in bladder cancer.
Fig. 2: SHMT2 promotes the proliferation, migration, and invasion of BC cells.
Fig. 3: SHMT2 is essential to one-carbon units for cell cycle progression.
Fig. 4: SHMT2 deficiency leads to increased ROS levels and mitochondrial apoptosis.
Fig. 5: SHMT2 deficiency induces ROS-dependent mitochondrial-mediated apoptosis.
Fig. 6: SHIN1 inhibits the proliferation, migration, and invasion of BC cells.
Fig. 7: SHIN1 induces ROS-dependent cell apoptosis via the intrinsic signaling pathway.

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All data generated and analyzed during this study are included in this article and its supplementary information files.

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Acknowledgements

We are grateful to Tong Zhao for the flow cytometry analysis and Shuo Wang for the critical reading of the manuscript and recommendations. This work was supported by grants from the Innovation Academy for Strategic Priority Research Program of the Chinese Academy of Sciences (XDA17010503), the National Natural Science Foundation of China (32071460), and the Innovation Academy for Green Manufacture, Chinese Academy of Sciences (IAGM-2019-A02).

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Authors and Affiliations

  1. CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 100101, Beijing, China

    Yun Zhang, Zhe Liu, Xueliang Wang, Hui Jian, Haihan Xiao & Tingyi Wen

  2. Innovation Academy for Green Manufacture, Chinese Academy of Sciences, 100190, Beijing, China

    Yun Zhang & Tingyi Wen

  3. University of Chinese Academy of Sciences, 100049, Beijing, China

    Zhe Liu, Xueliang Wang, Hui Jian & Haihan Xiao

  4. Savaid Medical School, University of Chinese Academy of Sciences, 100049, Beijing, China

    Tingyi Wen

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YZ and ZL designed and conducted the experiments; XLW aided for metabonomic data analysis and Figure drawing; HJ and HHX aided for experiment coordination; YZ analyzed the data and wrote the manuscript; TYW conceived of and supervised the project, analyzed data, made Figures, and revised the paper; and all authors read and approved the final manuscript.

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Correspondence to Tingyi Wen.

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Zhang, Y., Liu, Z., Wang, X. et al. SHMT2 promotes cell viability and inhibits ROS-dependent, mitochondrial-mediated apoptosis via the intrinsic signaling pathway in bladder cancer cells. Cancer Gene Ther 29, 1514–1527 (2022). https://doi.org/10.1038/s41417-022-00470-5

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