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Identification and characterization of colorectal-cancer-associated SNPs on the SMAD7 locus

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Abstract

Purpose

Genome-wide association studies have identified SMAD7 as a colorectal cancer (CRC) susceptibility gene. However, its underlying mechanism has not yet been characterized. This study screened functional SNPs (fSNPs) related to colorectal cancer through Reel-seq and obtained regulatory proteins on functional SNPs.

Methods

The candidate fSNPs on the SMAD7 locus were screened by Reel-seq method. Eight SNPs such as rs8085824 were identified as functional SNPs by luciferase reporter assay and EMSA, SDCP-MS and AIDP-WB revealed that HNRNPK can specifically bind to the rs8085824-C allele. The knockdown of HNRNPK by RNAi proved that HNRNPK could affect cell function by regulating SMAD7.

Results

Eight functional SNPs was found on the SMAD7 locus in linkage disequilibrium (LD) with R2 > 0.8, i.e., rs12953717, rs7227023, rs34007497, rs58920878, rs8085824, rs4991143, rs4939826, and rs7227023. We also identified allele-imbalanced binding of HNRNPK to rs8085824, H1-3 to rs12953717, THOC6 to rs7227023, and DDX21 to rs58920878. Further functional analysis revealed that these proteins are the regulatory proteins that modulate the expression of SMAD7 in the human colorectal cancer cell line DLD1. In particular, we discovered that siRNA knockdown of HNRNPK inhibits cell proliferation and cell clonal formation by downregulating SMAD7, as the decreased cell proliferation and cell clonal formation in the siRNA HNRNPK knockdown cells was restored by SMAD7 overexpression.

Conclusion

Our findings reveal a mechanism which underlies the contribution of the fSNP rs8085824 on the SMD7 locus to CRC susceptibility.

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Data availability

The data underlying this article are available in the article and in its online supplementary material.

References

  • Bogaert J, Prenen HJAOG (2014) Molecular genetics of colorectal cancer. Ann Gastroenterol 27(1):9–14

    PubMed  PubMed Central  Google Scholar 

  • Boulay JL, Mild G, Lowy A, Reuter J, Lagrange M, Terracciano L et al (2003) SMAD7 is a prognostic marker in patients with colorectal cancer. Int J Cancer 104:446–449

    Article  CAS  PubMed  Google Scholar 

  • Cheng X, Zhang F, Gong J, Li Y, Zhou D, Wang J et al (2022) Identification of potential functional variants and genes at 18q211 associated with the carcinogenesis of colorectal cancer. PLoS Genet 18:e1010050

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • De Simone V, Bevivino G, Sedda S, Izzo R, Laudisi F, Dinallo V et al (2017) Smad7 knockdown activates protein kinase RNA-associated eIF2α pathway leading to colon cancer cell death. Cell Death Dis 8:e2681–e2681

    Article  PubMed  PubMed Central  Google Scholar 

  • Edlund S, Lee SY, Grimsby S, Zhang S, Aspenström P, Heldin C-H et al (2005) Interaction between Smad7 and β-catenin: importance for transforming growth factor β-induced apoptosis. Mol Cell Biol 25:1475–1488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • El-Fattah A, Ahmed A, Sadik NAH, Shaker OG, Mohamed Kamal AJMOI (2018) Single nucleotide polymorphism in SMAD7 and CHI3L1 and colorectal cancer risk. 2018:9853192

  • Fitzpatrick-Lewis D, Ali MU, Warren R, Kenny M, Sherifali D, Raina PJCCC (2016) Screening for colorectal cancer: a systematic review and meta-analysis. Clin Colorectal Cancer 15:298–313

    Article  PubMed  Google Scholar 

  • Fortini BK, Tring S, Plummer SJ, Edlund CK, Moreno V, Bresalier RS et al (2014) Multiple functional risk variants in a SMAD7 enhancer implicate a colorectal cancer risk haplotype. PLoS ONE 9:e111914

    Article  PubMed  PubMed Central  Google Scholar 

  • Halder SK, Beauchamp RD, Datta PKJECR (2005) Smad7 induces tumorigenicity by blocking TGF-β-induced growth inhibition and apoptosis. Exp Cell Res 307:231–246

    Article  CAS  PubMed  Google Scholar 

  • Huang Y, Wu W, Nie M, Li C, Wang LJO (2016) SMAD7 polymorphisms and colorectal cancer risk: a meta-analysis of case–control studies. Oncotarget 7:75561

    Article  PubMed  PubMed Central  Google Scholar 

  • Keum N, Giovannucci EJN, r. G. & hepatology. (2019) Global burden of colorectal cancer: emerging trends, risk factors and prevention strategies. Nat Rev Gastroenterol Hepatol 16:713–732

    Article  PubMed  Google Scholar 

  • Liu S, Liu Y, Zhang Q, Wu J, Liang J, Yu S et al (2017) Systematic identification of regulatory variants associated with cancer risk. Genome Biol 18:1–14

    Article  Google Scholar 

  • Nakao AJIOS, Afrakhte M, Moren A, Nakayama T, Christian JL, Heuchel R, Itoh S, Kawabata M, Heldin NE, Heldin CH, Ten Dijke P (1997) A TGFbeta-inducible antagonist of TGF-beta signalling. Nature 389:631–635

    Article  CAS  PubMed  Google Scholar 

  • Rao H-L, Chen J-W, Li M, Xiao Y-B, Fu J, Zeng Y-X et al (2012) Increased intratumoral neutrophil in colorectal carcinomas correlates closely with malignant phenotype and predicts patients’ adverse prognosis. PLoS ONE 7:e30806

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Rich JNJCR (2007) Cancer stem cells in radiation resistance. Cancer Res 67:8980–8984

    Article  CAS  PubMed  Google Scholar 

  • Sanjana NE, Shalem O, Zhang FJNM (2014) Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods 11:783–784

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stolfi C, De Simone V, Colantoni A, Franze E, Ribichini E, Fantini M et al (2014) A functional role for Smad7 in sustaining colon cancer cell growth and survival. Cell Death Dis 5:e1073–e1073

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Tong L, Shen S, Huang Q, Fu J, Wang T, Pan L et al (2020) Proteasome-dependent degradation of Smad7 is critical for lung cancer metastasis. Cell Death Differ 27:1795–1806

    Article  CAS  PubMed  Google Scholar 

  • Venables JP, Koh C-S, Froehlich U, Lapointe E, Couture S, Inkel L et al (2008) Multiple and specific mRNA processing targets for the major human hnRNP proteins. Mol Cell Biol 28:6033–6043

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang Z, Qiu H, He J, Liu L, Xue W, Fox A et al (2020) The emerging roles of hnRNPK. J Cell Physiol 235:1995–2008

    Article  CAS  PubMed  Google Scholar 

  • Zhao Y, Zhao M-F, Jiang S, Wu J, Liu J, Yuan X-W et al (2020) Liver governs adipose remodelling via extracellular vesicles in response to lipid overload. Nat Commun 11:719

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Dr. Gang Li (University of Pittsburgh, USA) for scientific discussions concerning this work.

Funding

This work was supported by Science and Technology Commission of Shanghai Municipality (23141901800, 23141903300) and the National Nature Science Foundation of China (82173099, 32270960).

Author information

Authors and Affiliations

  1. Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai, 200241, China

    Zhao Liu, Yihan Zhao, Hongli Song, Huaxue Miao, Yingying Wang, Chuntian Tu, Juliang Qin, Bing Du, Min Qian & Hua Ren

  2. School of Mathematical Sciences, East China Normal University, Shanghai, 200241, China

    Tianyun Fu

  3. Joint Center for Translational Medicine, Fengxian District Central Hospital, Fengxian District, Shanghai, 201499, China

    Juliang Qin

Authors
  1. Zhao Liu
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  2. Yihan Zhao
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  3. Hongli Song
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  4. Huaxue Miao
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  5. Yingying Wang
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  6. Chuntian Tu
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  7. Tianyun Fu
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  9. Bing Du
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  10. Min Qian
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  11. Hua Ren
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Contributions

MQ and HR supervised the project. ZL, HS, HM, YW and CT performed the experiments and analyzed the data. MQ, YZ and TF supervised the Reel-seq experiments. BD and JQ provided advice and technical expertise. HR and ZL wrote the manuscript. All authors revised the manuscript and approved its final version.

Corresponding authors

Correspondence to Min Qian or Hua Ren.

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Conflict of interest

The authors declare no relevant financial interests and no competing interest.

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Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 Supplementary Figure 1. HNRNPK binding sequence comparison with rs8085824 (TIF 2505 KB)

432_2023_5402_MOESM2_ESM.tif

Supplementary file2 Supplementary Figure 2. The luciferase reporter gene demonstrated that HNRNPK could regulate luciferase activity through rs8085824-C, through knockdown or overexpression of HNRNPK in DLD1 cells(n>3). RLA relative luciferase activity, P value was calculated using Student’s t test with two tails. Error bars, SD. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (TIF 4403 KB)

Supplementary file3 (DOCX 27 KB)

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Liu, Z., Zhao, Y., Song, H. et al. Identification and characterization of colorectal-cancer-associated SNPs on the SMAD7 locus. J Cancer Res Clin Oncol 149, 16659–16668 (2023). https://doi.org/10.1007/s00432-023-05402-w

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