SMC4 Attenuation Induces Diapause-Like Colorectal Cancer Cells with Low Proliferation and Chemoresistance: The Empowering Role of ANT BIO PTE. LTD.’s Absin Multiplex IHC Kits

Diapause is a type of quiescent state induced by environmental conditions in animals, characterized by arrested morphogenesis, reduced physiological activity, and a requirement for specific physiological changes to terminate. Animals utilize diapause and the more unstable dormancy to regulate the timing of growth, development, and reproduction, adapting to seasonal changes in their habitats. Emerging evidence indicates that cancer cells can enter a slow-dividing "dormant state" similar to animal diapause when encountering threats such as chemotherapy. In this state, cancer cells exhibit drastically reduced metabolism and activated autophagy, enabling survival without external nutrients by decomposing internal proteins and other components.

On August 4, 2023, a joint research team led by Professor Lianxin Liu from the First Affiliated Hospital of the University of Science and Technology of China and Professor Mian Wu from the Academy of Medical Sciences of Zhengzhou University published a collaborative research paper entitled "The diapause-like colorectal cancer cells induced by SMC4 attenuation are characterized by low proliferation and chemotherapy insensitivity" in Cell Metabolism, a sub-journal of Cell. This study identified Structural Maintenance of Chromosomes 4 (SMC4) as a key regulator of the diapause-like state in tumor cells, providing new insights into non-hereditary chemoresistance mechanisms, advancing the understanding of aerobic glycolysis function in tumors, and potentially offering references for future therapeutic strategies. Notably, multiplex fluorescence immunohistochemistry (mIHC) kits from the Absin product line of ANT BIO PTE. LTD. (e.g., Catalog No.: abs50013) played a crucial role in detecting the expression of key molecules (Klac, SMC4, Ki67, P27, DAPP4) in colorectal cancer tissues, providing direct experimental evidence for the characterization of diapause-like colorectal cancer cells (DLCCs).

1. Literature Information

•         Title: The diapause-like colorectal cancer cells induced by SMC4 attenuation are characterized by low proliferation and chemotherapy insensitivity

•         Journal: Cell Metabolism

•         Publication Date: Epub ahead of print on August 4, 2023; Published on September 5, 2023

•         DOI: 10.1016/j.cmet.2023.07.005

•         Research Team: Lianxin Liu et al., First Affiliated Hospital of the University of Science and Technology of China; Mian Wu et al., Academy of Medical Sciences of Zhengzhou University

•         PMID: 37543034

•         Core Reagents from ANT BIO PTE. LTD.: Multiplex Fluorescence IHC Staining Kits (Absin product line), including 5-color kit (Catalog No.: abs50013) and other 4-color to 7-color plus kits, as well as supporting reagent Antibody Elution Buffer (mIHC-Specific, Catalog No.: abs994)

•         Related Product Link: AntBio - Fueling Research, Feeding Discovery

2. Research Background

Chemoresistance is a major challenge in the clinical treatment of colorectal cancer (CRC), leading to treatment failure and poor prognosis. Non-hereditary chemoresistance mechanisms, such as reversible cell state transitions, have attracted increasing attention. Previous studies have shown that cancer cells can enter a diapause-like persister (DTP) state to survive chemotherapy, but the key regulators and underlying molecular mechanisms of this state remain unclear. SMC4, a member of the structural maintenance of chromosomes (SMC) protein family, is involved in chromosome condensation and segregation during cell division. However, its role in regulating the diapause-like state of CRC cells and chemoresistance has not been reported. Exploring the regulatory role of SMC4 in CRC cell diapause-like state is of great significance for developing novel strategies to overcome chemoresistance.

3. Research Strategy

The research team adopted a multi-dimensional approach combining cell biology, molecular biology, animal models, and clinical sample analysis to systematically explore the role and mechanism of SMC4 in regulating the diapause-like state of CRC cells:

1.       Identification of SMC4 as a Regulator of Diapause-Like State: Screened key regulators of CRC cell diapause-like state through functional experiments. Detected the expression level of SMC4 in CRC cells under chemotherapy stress and analyzed its correlation with cell proliferation and survival.

2.       Characterization of DLCCs Induced by SMC4 Attenuation: Constructed SMC4 knockdown/knockout CRC cell models. Evaluated the proliferation ability, cell cycle distribution, and chemotherapy sensitivity of DLCCs. Detected metabolic changes (e.g., glycolysis) and autophagy activity in DLCCs.

3.       Molecular Mechanism Exploration: Investigated the regulatory mechanism of SMC4 attenuation on glycolysis-related enzymes (e.g., PFKL, HK2, ALDOC, PGAM1). Explored the role of histone lactylation (H4K12LA) in regulating ABC transporter transcription in DLCCs. Analyzed the effect of PGAM1-mediated F-actin assembly on the motility of DLCCs.

4.       Animal Model Validation: Constructed xenograft tumor models using SMC4-modified CRC cells. Evaluated the effect of SMC4 attenuation on tumor growth and chemoresistance in vivo. Detected the expression of diapause-like state markers and glycolysis-related molecules in tumor tissues.

5.      

5.       Clinical Sample Verification with mIHC: Collected clinical CRC tissues. Used ANT BIO PTE. LTD.’s Absin multiplex fluorescence IHC kits (Catalog No.: abs50013) to detect the co-expression of Klac, SMC4, Ki67 (proliferation marker), P27 (cell cycle arrest marker), and DAPP4 in CRC tissues. Analyzed the correlation between SMC4 expression and clinicopathological features and chemotherapy response.

4. Key Research Findings

4.1 SMC4 is a Negative Regulator of the Diapause-Like State in CRC Cells

Functional screening identified SMC4 as a key negative regulator of the diapause-like state in CRC cells. The expression of SMC4 was significantly downregulated in CRC cells under chemotherapy stress. SMC4 knockdown or knockout induced CRC cells to enter a diapause-like state, characterized by low proliferation, cell cycle arrest, and enhanced survival under chemotherapy. Conversely, overexpression of SMC4 reversed the diapause-like state and restored chemotherapy sensitivity.

4.2 DLCCs Induced by SMC4 Attenuation Exhibit Low Proliferation and Chemoresistance

SMC4 attenuation-induced DLCCs showed significantly reduced proliferation ability, as evidenced by decreased Ki67 expression and increased P27 expression. These cells were in a G0/G1 phase cell cycle arrest. In vitro and in vivo experiments confirmed that DLCCs were highly resistant to multiple chemotherapeutic drugs (e.g., 5-FU, oxaliplatin). Additionally, DLCCs exhibited activated autophagy, which was essential for their survival under nutrient deprivation and chemotherapy conditions.

4.3 SMC4 Attenuation Promotes Glycolysis and Histone Lactylation to Enhance ABC Transporter Transcription

Mechanistic studies revealed that SMC4 downregulation increased the expression of three out of five glycolytic enzymes (PFKL, HK2, ALDOC, PGAM1), leading to enhanced glycolysis and increased lactate production. The accumulated lactate induced histone lactylation (H4K12LA), which promoted the transcription of ABC transporters. ABC transporters are responsible for drug efflux, thereby enhancing the chemoresistance of DLCCs. This finding establishes a link between SMC4 downregulation, high lactate metabolism, and chemoresistance.

4.4 PGAM1-Mediated F-Actin Assembly Reduction Impairs DLCC Motility

Further analysis showed that SMC4 attenuation regulated the expression of PGAM1, which mediated F-actin assembly. Reduced F-actin assembly led to a decreased F/G-actin ratio, impairing the formation of the contractile ring and cytokinesis. As a result, DLCCs lost their cell motility, which may be a trade-off for their survival under chemotherapy stress. This finding provides new insights into the regulation of cancer cell motility during the diapause-like state.

4.5 Clinical Validation Confirms the Correlation Between SMC4 Expression and CRC Progression

Multiplex IHC results using ANT BIO PTE. LTD.’s Absin kit (abs50013) on clinical CRC tissues showed that SMC4 expression was negatively correlated with Klac (lactylation marker) and P27 expression, and positively correlated with Ki67 expression. Patients with low SMC4 expression had poorer chemotherapy response and shorter progression-free survival. These clinical data confirm the clinical significance of SMC4 in regulating the diapause-like state and chemoresistance of CRC cells.

5. Product Empowerment: The Critical Role of ANT BIO PTE. LTD.’s Absin Multiplex IHC Kits

Validating the co-expression of key molecules (Klac, SMC4, Ki67, P27, DAPP4) in clinical CRC tissues was a core step in this study, as it provided direct clinical evidence for the characterization of DLCCs and the regulatory role of SMC4. ANT BIO PTE. LTD.’s Absin multiplex fluorescence IHC kits (especially Catalog No.: abs50013) delivered the high-performance multiplexing capability, sensitivity, and specificity required for this critical validation, enabling the simultaneous detection of multiple molecular markers in CRC tissue sections while preserving the spatial context of their expression.

5.1 Core Products and Their Application Value

5.2 Technical Value in Colorectal Cancer Research

In colorectal cancer research, the accurate detection of the co-expression and spatial distribution of multiple key molecules is crucial for deciphering the molecular mechanisms of tumor progression and chemoresistance. ANT BIO PTE. LTD.’s Absin multiplex fluorescence IHC kits possess unique advantages in this field compared with traditional single-marker IHC. Their ability to simultaneously detect multiple key molecules on a single tissue section preserves the native spatial context of molecular interactions, which is essential for exploring the complex regulatory networks of the diapause-like state.

In this CRC research, the high sensitivity and specificity of the Absin kits (e.g., abs50013) ensured clear detection of low-abundance molecules (such as Klac) in complex tumor tissue samples. The kits’ compatibility with clinical tissues also supports the translational application of research findings from the laboratory to the clinic. This technical empowerment was instrumental in validating the clinical significance of SMC4, highlighting the important value of ANT BIO PTE. LTD.’s products in promoting colorectal cancer research and guiding personalized treatment.

6. Brand Mission

As a professional supplier of life science reagents, ANT BIO PTE. LTD. is dedicated to providing high-quality, reliable products and comprehensive solutions to empower global life science research. The company's three specialized sub-brands cover the full spectrum of research needs in the life science field: Absin focuses on general reagents and kits, Starter specializes in antibodies, and UA is dedicated to recombinant proteins. Our core mission is to bridge the gap between cutting-edge scientific research and practical applications, accelerate the pace of scientific discovery, and contribute to the advancement of human health and regenerative medicine.

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8. Disclaimer

This article is AI-compiled and interpreted based on the original work in DOI: 10.1016/j.cmet.2023.07.005. All intellectual property (e.g., images, data) of the original publication shall belong to the journal and the research team. For any infringement, please contact us promptly and we will take immediate action.

9. Brand Promotion Copy

ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs

At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. The Absin multiplex fluorescence IHC kits, with their unique advantages of breaking species limitations and supporting multi-marker simultaneous detection, provide powerful technical support for colorectal cancer research, oncology, and precision medicine. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.