Skip to main content

Advertisement

SpringerLink
Log in

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

  • RESEARCH ARTICLE
  • Published:
Clinical and Translational Oncology Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

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

References

  1. Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2021. Cancer J Clin. 2021;71(1):7–33.

    Article  Google Scholar 

  2. Marquardt S, Solanki M, Spitschak A, Vera J, Putzer BM. Emerging functional markers for cancer stem cell-based therapies: understanding signaling networks for targeting metastasis. Semin Cancer Biol. 2018;53:90–109.

    Article  CAS  PubMed  Google Scholar 

  3. Di Franco S, Bianca P, Sardina DS, Turdo A, Gaggianesi M, Veschi V, et al. Adipose stem cell niche reprograms the colorectal cancer stem cell metastatic machinery. Nat Commun. 2021;12(1):5006.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Najafi M, Farhood B, Mortezaee K. Cancer stem cells (CSCs) in cancer progression and therapy. J Cell Physiol. 2019;234(6):8381–95.

    Article  CAS  PubMed  Google Scholar 

  5. Dianat-Moghadam H, Heidarifard M, Jahanban-Esfahlan R, Panahi Y, Hamishehkar H, Pouremamali F, et al. Cancer stem cells-emanated therapy resistance: Implications for liposomal drug delivery systems. J Controll Releas : Off J Controll Releas Soc. 2018;288:62–83.

    Article  CAS  Google Scholar 

  6. Jarrett SG, Wolf Horrell EM, D’Orazio JA. Retraction: AKAP12 mediates PKA-induced phosphorylation of ATR to enhance nucleotide excision repair. Nucleic Acids Res. 2020;48(20):11814.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Wu X, Luo Y, Wang S, Li Y, Bao M, Shang Y, et al. AKAP12 ameliorates liver injury via targeting PI3K/AKT/PCSK6 pathway. Redox Biol. 2022;53: 102328.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Liu W, Guan M, Hu T, Gu X, Lu Y. Re-expression of AKAP12 inhibits progression and metastasis potential of colorectal carcinoma in vivo and in vitro. PLoS ONE. 2011;6(8): e24015.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Suren D, Yildirim M, Alikanoglu AS, Kaya V, Yildiz M, Dilli UD, et al. Lack of relation of AKAP12 with p53 and Bcl-2 in colorectal carcinoma. Asian Pac J Cancer Prev : APJCP. 2014;15(8):3415–8.

    Article  PubMed  Google Scholar 

  10. Su B, Bu Y, Engelberg D, Gelman IH. SSeCKS/Gravin/AKAP12 inhibits cancer cell invasiveness and chemotaxis by suppressing a protein kinase C- Raf/MEK/ERK pathway. J Biol Chem. 2010;285(7):4578–86.

    Article  CAS  PubMed  Google Scholar 

  11. Lee SW, Jung KH, Jeong CH, Seo JH, Yoon DK, Suh JK, et al. Inhibition of endothelial cell migration through the downregulation of MMP-9 by A-kinase anchoring protein 12. Mol Med Rep. 2011;4(1):145–9.

    CAS  PubMed  Google Scholar 

  12. Yang JM, Lee HS, Seo JH, Park JH, Gelman IH, Lo EH, et al. Structural environment built by AKAP12+ colon mesenchymal cells drives M2 macrophages during inflammation recovery. Sci Rep. 2017;7:42723.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Bots M, Verbrugge I, Martin BP, Salmon JM, Ghisi M, Baker A, et al. Differentiation therapy for the treatment of t (8; 21) acute myeloid leukemia using histone deacetylase inhibitors. Blood. 2014;123(9):1341–52.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Rasanen K, Lehtinen E, Nokelainen K, Kuopio T, Hautala L, Itkonen O, et al. Interleukin-6 increases expression of serine protease inhibitor Kazal type 1 through STAT3 in colorectal adenocarcinoma. Mol Carcinog. 2016;55(12):2010–23.

    Article  PubMed  Google Scholar 

  15. Heichler C, Scheibe K, Schmied A, Geppert CI, Schmid B, Wirtz S, et al. STAT3 activation through IL-6/IL-11 in cancer-associated fibroblasts promotes colorectal tumour development and correlates with poor prognosis. Gut. 2020;69(7):1269–82.

    Article  CAS  PubMed  Google Scholar 

  16. Venkatesan N, Wong JF, Tan KP, Chung HH, Yau YH, Cukuroglu E, et al. EZH2 promotes neoplastic transformation through VAV interaction-dependent extranuclear mechanisms. Oncogene. 2017. https://doi.org/10.1038/onc.2017.309.

    Article  PubMed  Google Scholar 

  17. Wang W, Guo C, Zhu P, Lu J, Li W, Liu C, et al. Phosphorylation of STAT3 mediates the induction of cyclooxygenase-2 by cortisol in the human amnion at parturition. Sci Signal. 2015;8(400):ra106.

    Article  PubMed  Google Scholar 

  18. Turtoi A, Mottet D, Matheus N, Dumont B, Peixoto P, Hennequiere V, et al. The angiogenesis suppressor gene AKAP12 is under the epigenetic control of HDAC7 in endothelial cells. Angiogenesis. 2012;15(4):543–54.

    Article  CAS  PubMed  Google Scholar 

  19. Mostafa MR, Yahia RS, Abd El Messih HM, El-Sisy E, El Ghannam DM. Gravin gene expression in acute myeloid leukemia. Med Oncol (Northwood, London, England). 2013;30(2):548.

    Article  Google Scholar 

  20. Peixoto P, Blomme A, Costanza B, Ronca R, Rezzola S, Palacios AP, et al. HDAC7 inhibition resets STAT3 tumorigenic activity in human glioblastoma independently of EGFR and PTEN: new opportunities for selected targeted therapies. Oncogene. 2016;35(34):4481–94.

    Article  CAS  PubMed  Google Scholar 

  21. Jo U, Whang YM, Kim HK, Kim YH. AKAP12alpha is associated with promoter methylation in lung cancer. Cancer Res Treat : Off J Korean Cancer Assoc. 2006;38(3):144–51.

    Article  Google Scholar 

  22. Cosenza M, Civallero M, Pozzi S, Marcheselli L, Bari A, Sacchi S. The combination of bortezomib with enzastaurin or lenalidomide enhances cytotoxicity in follicular and mantle cell lymphoma cell lines. Hematol Oncol. 2015;33(4):166–75.

    Article  CAS  PubMed  Google Scholar 

  23. Shaheen S, Ahmed M, Lorenzi F, Nateri AS. Spheroid-formation (Colonosphere) assay for in vitro assessment and expansion of stem cells in colon cancer. Stem Cell Rev. 2016;12(4):492–9.

    Article  CAS  PubMed Central  Google Scholar 

  24. Yang Z, He L, Lin K, Zhang Y, Deng A, Liang Y, et al. The KMT1A-GATA3-STAT3 Circuit is a novel self-renewal signaling of human bladder cancer stem cells. Clin Cancer Res : Off J Am Assoc Cancer Res. 2017. https://doi.org/10.1158/1078-0432.CCR-17-0882.

    Article  Google Scholar 

  25. Shien K, Papadimitrakopoulou VA, Ruder D, Behrens C, Shen L, Kalhor N, et al. JAK1/STAT3 Activation through a proinflammatory cytokine pathway leads to resistance to molecularly targeted therapy in non-small cell lung cancer. Mol Cancer Ther. 2017;16(10):2234–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Hatano Y, Fukuda S, Hisamatsu K, Hirata A, Hara A, Tomita H. Multifaceted interpretation of colon cancer stem cells. Int J Mol Sci. 2017. https://doi.org/10.3390/ijms18071446.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Wang R, Wei J, Zhang S, Wu X, Guo J, Liu M, et al. Peroxiredoxin 2 is essential for maintaining cancer stem cell-like phenotype through activation of Hedgehog signaling pathway in colon cancer. Oncotarget. 2016;7(52):86816–28.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Najafi M, Mortezaee K, Majidpoor J. Cancer stem cell (CSC) resistance drivers. Life Sci. 2019;234: 116781.

    Article  CAS  PubMed  Google Scholar 

  29. Srivastava AK, Banerjee A, Cui T, Han C, Cai S, Liu L, et al. Inhibition of miR-328-3p impairs cancer stem cell function and prevents metastasis in ovarian cancer. Can Res. 2019;79(9):2314–26.

    Article  CAS  Google Scholar 

  30. Wang P, Wan WW, Xiong SL, Feng H, Wu N. Cancer stem-like cells can be induced through dedifferentiation under hypoxic conditions in glioma, hepatoma and lung cancer. Cell Death Discov. 2017;3:16105.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Zeuner A, Todaro M, Stassi G, De Maria R. Colorectal cancer stem cells: from the crypt to the clinic. Cell Stem Cell. 2014;15(6):692–705.

    Article  CAS  PubMed  Google Scholar 

  32. Soheilifar MH, Moshtaghian A, Maadi H, Izadi F, Saidijam M. BMI1 Roles in cancer stem cells and its association with MicroRNAs dysregulation in cancer: emphasis on colorectal cancer. Int J Cancer Manag. 2018. https://doi.org/10.5812/ijcm.82926.

    Article  Google Scholar 

  33. Zhu Y, Huang S, Chen S, Chen J, Wang Z, Wang Y, et al. SOX2 promotes chemoresistance, cancer stem cells properties, and epithelial-mesenchymal transition by β-catenin and Beclin1/autophagy signaling in colorectal cancer. Cell Death Dis. 2021;12(5):449.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Zhang M, Peng R, Wang H, Yang Z, Zhang H, Zhang Y, et al. Nanog mediated by FAO/ACLY signaling induces cellular dormancy in colorectal cancer cells. Cell Death Dis. 2022;13(2):159.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Yan X, Liu L, Li H, Qin H, Sun Z. Clinical significance of Fusobacterium nucleatum, epithelial-mesenchymal transition, and cancer stem cell markers in stage III/IV colorectal cancer patients. Onco Targets Ther. 2017;10:5031–46.

    Article  PubMed  PubMed Central  Google Scholar 

  36. Liu W, Guan M, Su B, Ye C, Li J, Zhang X, et al. Quantitative assessment of AKAP12 promoter methylation in colorectal cancer using methylation-sensitive high resolution melting: correlation with Duke’s stage. Cancer Biol Ther. 2010;9(11):862–71.

    Article  CAS  PubMed  Google Scholar 

  37. Maki T, Choi YK, Miyamoto N, Shindo A, Liang AC, Ahn BJ, et al. A-kinase anchor protein 12 is required for oligodendrocyte differentiation in adult white matter. Stem Cells (Dayton, Ohio). 2018. https://doi.org/10.1002/stem.2771.

    Article  PubMed  Google Scholar 

  38. Deng Y, Gao J, Xu G, Yao Y, Sun Y, Shi Y, et al. HDAC6-dependent deacetylation of AKAP12 dictates its ubiquitination and promotes colon cancer metastasis. Cancer Lett. 2022;549: 215911.

    Article  CAS  PubMed  Google Scholar 

  39. Liu W, Kovacevic Z, Peng Z, Jin R, Wang P, Yue F, et al. The molecular effect of metastasis suppressors on Src signaling and tumorigenesis: new therapeutic targets. Oncotarget. 2015;6(34):35522–41.

    Article  PubMed  PubMed Central  Google Scholar 

  40. Akakura S, Gelman IH. Pivotal role of AKAP12 in the regulation of cellular adhesion dynamics: control of cytoskeletal architecture, cell migration, and mitogenic signaling. J Sign Transduct. 2012;2012: 529179.

    Google Scholar 

  41. Yao C, Su L, Shan J, Zhu C, Liu L, Liu C, et al. IGF/STAT3/NANOG/Slug signaling axis simultaneously controls epithelial-mesenchymal transition and stemness maintenance in colorectal cancer. Stem Cells. 2016;34(4):820–31.

    Article  PubMed  Google Scholar 

  42. Jin W. Role of JAK/STAT3 signaling in the regulation of metastasis, the transition of cancer stem cells, and chemoresistance of cancer by epithelial-mesenchymal transition. Cells. 2020. https://doi.org/10.3390/cells9010217.

    Article  PubMed  PubMed Central  Google Scholar 

  43. Lin L, Liu A, Peng Z, Lin HJ, Li PK, Li C, et al. STAT3 is necessary for proliferation and survival in colon cancer-initiating cells. Can Res. 2011;71(23):7226–37.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

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

Funding

Not applicable.

Author information

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

Authors
  1. Ke Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xuan Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yuan Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ting-Ting Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Weifeng Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Frank J. Gonzalez
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Weiwei Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Weiwei Liu.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

The research involving animals strictly abides by the ethical regulations of Longhua Hospital affiliated to Shanghai University of Traditional Chinese Medicine and has been approved by the ethics committee.

Consent for publication

Not applicable.

Informed consent statement

For this type of study, informed consent statement is not required.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

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)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

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

Keywords

Search

Navigation