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AKAP12 promotes cancer stem cell-like phenotypes and activates STAT3 in colorectal cancer

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Abstract

Background

Cancer stem cells (CSCs) have unique biological characteristics, including tumorigenicity, immortality, and chemoresistance. Colorectal CSCs have been identified and isolated from colorectal cancers by various methods. AKAP12, a scaffolding protein, is considered to act as a potential suppressor in colorectal cancer, but its role in CSCs remains unknown. In this study, we investigated the function of AKAP12 in Colorectal CSCs.

Methods

Herein, Colorectal CSCs were enriched by cell culture with a serum-free medium. CSC-associated characteristics were evaluated by Flow cytometry assay and qPCR. AKAP12 gene expression was regulated by lentiviral transfection assay. The tumorigenicity of AKAP12 in vivo by constructing a tumor xenograft model. The related pathways were explored by qPCR and Western blot.

Results

The depletion of AKAP12 reduced colony formation, sphere formation, and expression of stem cell markers in colorectal cancer cells, while its knockdown decreased the volume and weight of tumor xenografts in vivo. AKAP12 expression levels also affected the expression of stemness markers associated with STAT3, potentially via regulating the expression of protein kinase C.

Conclusion

This study suggests Colorectal CSCs overexpress AKAP12 and maintain stem cell characteristics through the AKAP12/PKC/STAT3 pathway. AKAP12 may be an important therapeutic target for blocking the development of colorectal cancer in the field of cancer stem cells.

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Data availability

The analyzed data sets generated during the study are available from the corresponding author on reasonable request.

Abbreviations

CSCs:

Cancer stem cells

CCSCs:

Colorectal cancer stem cells

AKAP12:

A-kinase anchor protein 12

STAT3:

Signal transduction and transcription activator 3

Lv-hAKAP12:

Lentiviruses overexpressing AKAP12

Lv-Null:

Negative control

Lv-siAKAP12:

Lentiviruses with AKAP12-siRNA

Lv-siCont:

Negative control

PKC:

Protein kinase C

qPCR:

Quantitative polymerase chain reaction

RT-PCR:

Reverse transcription PCR

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Acknowledgements

All authors sincerely thank the Central Laboratory of Shanghai Tenth People's Hospital for providing experimental equipment and experimental platform for this research.

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Authors and Affiliations

  1. Department of Laboratory Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, 200032, People’s Republic of China

    Ke Li, Xuan Wu, Weifeng Wang & Weiwei Liu

  2. Department of Laboratory Medicine, Shanghai Tenth People’s Hospital Affiliated to Tongji University, Shanghai, 200070, People’s Republic of China

    Xuan Wu & Yuan Li

  3. Department of Laboratory Medicine, Huashan Hospital, Shanghai Medical College, Fudan University, Shanghai, 200040, People’s Republic of China

    Ting-Ting Hu

  4. Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA

    Frank J. Gonzalez & Weiwei Liu

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  1. Ke Li
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Correspondence to Weiwei Liu.

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12094_2023_3230_MOESM1_ESM.tif

Sup. Fig. 1:The proportion of cells staying in S-phase and G2M phase was increased in HCT116-hAKAP12 cells, Supplementary file1 (TIF 1007 KB)

12094_2023_3230_MOESM2_ESM.tif

Sup. Fig. 2:The proportion of cells staying in S-phase and G2M phase was increased in LoVo-hAKAP12 cells, Supplementary file2 (TIF 124 KB)

12094_2023_3230_MOESM3_ESM.tif

Sup. Fig. 3:Statistical graph of the corresponding WB results in Figure 5. A-D correspond to Figure 5B, D, F and H respectively, Supplementary file3 (TIF 201 KB)

Supplementary file4 (DOCX 80 KB)

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Li, K., Wu, X., Li, Y. et al. AKAP12 promotes cancer stem cell-like phenotypes and activates STAT3 in colorectal cancer. Clin Transl Oncol 25, 3263–3276 (2023). https://doi.org/10.1007/s12094-023-03230-5

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  • DOI: https://doi.org/10.1007/s12094-023-03230-5

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