An Integrated Analysis of NF-κB, MMP-9 Expression, and Tumor-Infiltrating Lymphocytes Density in Correlation with TNM Stage of Colorectal Cancer

ORIGINAL ARTICLE
Heru Fajar Trianto, Gondo Mastutik, Desak Gede Agung Suprabawati, Mitra Handini, Mahyarudin Mahyarudin

 
For citation: Trianto HF, Mastutik G, Suprabawati DGA, Handini M, Mahyarudin M. An Integrated Analysis of NF-κB, MMP-9 Expression, and Tumor-Infiltrating Lymphocytes Density in Correlation with TNM Stage of Colorectal Cancer. International Journal of Biomedicine. 2026;16(2):163-168. doi:10.21103/Article16(2)_OA2
 
Originally published June 5, 2026

Abstract: 

Background: Colorectal cancer progression is influenced by interactions between tumor cells and the stromal area. Nuclear factor kappa B (NF-κB) and matrix metalloproteinase-9 (MMP-9) contribute to cancer expansion, while tumor-infiltrating lymphocyte (TIL) density influences the host immune response. However, their complexity remains unclear. Therefore, this study aimed to integrate an analysis of the correlation between NF-κB, MMP-9, and TIL density with TNM stage in colorectal cancer.
Methods and Results: Fifty colorectal cancer paraffin blocks were analyzed. NF-κB and MMP-9 expression were evaluated by immunohistochemistry, while TIL density was assessed using hematoxylin-eosin (HE) staining. Correlations with TNM stage and its components were analyzed using Spearman’s correlation test. NF-κB expression correlated positively with N stage (p=0.013). MMP-9 expression had a positive correlation with T stage (p=0.000), M stage (p=0.036), and TNM stage (p=0.001). In contrast, TIL density showed an inverse correlation with T stage (p=0.010), N stage (p=0.003), and TNM stage (p=0.001). NF-κB expression correlated positively with MMP-9 (p=0.021), but not with TIL density.
Conclusion: NF-κB and MMP-9 expressions contribute to progression, while TIL density contributes to the antitumor response in colorectal cancer. An integrated analysis of the biomarkers showed a dynamic interaction between tumor molecular changes and the host immune response.

Keywords: 
transcription factor • matrix metalloproteinase • immune response • cancer
References: 
  1. Roshandel G, Ghasemi-Kebria F, Malekzadeh R. Colorectal Cancer: Epidemiology, Risk Factors, and Prevention. Cancers (Basel). 2024 Apr 17;16(8):1530. doi: 10.3390/cancers16081530. PMID: 38672612; PMCID: PMC11049480.
  2. Weiser MR. AJCC 8th Edition: Colorectal Cancer. Ann Surg Oncol. 2018 Jun;25(6):1454-1455. doi: 10.1245/s10434-018-6462-1. Epub 2018 Apr 3. PMID: 29616422.
  3. Fleming M, Ravula S, Tatishchev SF, Wang HL. Colorectal carcinoma: Pathologic aspects. J Gastrointest Oncol. 2012 Sep;3(3):153-73. doi: 10.3978/j.issn.2078-6891.2012.030. PMID: 22943008; PMCID: PMC3418538.
  4. Coebergh van den Braak RRJ, Ten Hoorn S, Sieuwerts AM, Tuynman JB, Smid M, Wilting SM, et al. Interconnectivity between molecular subtypes and tumor stage in colorectal cancer. BMC Cancer. 2020 Sep 4;20(1):850. doi: 10.1186/s12885-020-07316-z. PMID: 32887573; PMCID: PMC7473811.
  5. Sadati S, Khalaji A, Bonyad A, Khoshdooz S, Hosseini Kolbadi KS, Bahrami A, et al. NF-κB and apoptosis: colorectal cancer progression and novel strategies for treatment. Eur J Med Res. 2025 Jul 14;30(1):616. doi: 10.1186/s40001-025-02734-w. PMID: 40660346; PMCID: PMC12261797.
  6. Lukas K, Nguyen J, Necas C, Dave K, Venketaraman V. Targeting the NF-κB Pathway in Cancer: Mechanisms, Resistance, and Therapeutic Potential Across Tumor Types. Pharmaceuticals (Basel). 2025 Nov 20;18(11):1764. doi: 10.3390/ph18111764. PMID: 41305005; PMCID: PMC12655786.
  7. Mao H, Zhao X, Sun SC. NF-κB in inflammation and cancer. Cell Mol Immunol. 2025 Aug;22(8):811-839. doi: 10.1038/s41423-025-01310-w. Epub 2025 Jun 25. PMID: 40562870; PMCID: PMC12310982.
  8. Veljkovic A, Stanojevic G, Brankovic B, Roumeliotis S, Leivaditis K, Djordjevic B, et al. MMP-9 Activation via ROS/NF-κB Signaling in Colorectal Cancer Progression: Molecular Insights and Prognostic-Therapeutic Perspectives. Curr Issues Mol Biol. 2025 Jul 17;47(7):557. doi: 10.3390/cimb47070557. PMID: 40729026; PMCID: PMC12293130.
  9. Mudatsir, Labeda I, Uwuratuw JA, Hendarto J, Warsinggih, Lusikooy RE, et al. Relationship between metalloproteinase-9 (MMP-9) expression and clinicopathology in colorectal cancer: a cross-sectional study. Ann Med Surg (Lond). 2023 Jul 25;85(9):4277-4282. doi: 10.1097/MS9.0000000000000892. PMID: 37663709; PMCID: PMC10473300.
  10. Antonangeli F, Natalini A, Garassino MC, Sica A, Santoni A, Di Rosa F. Regulation of PD-L1 Expression by NF-κB in Cancer. Front Immunol. 2020 Nov 25;11:584626. doi: 10.3389/fimmu.2020.584626. PMID: 33324403; PMCID: PMC7724774.
  11. Bai Z, Zhou Y, Ye Z, Xiong J, Lan H, Wang F. Tumor-Infiltrating Lymphocytes in Colorectal Cancer: The Fundamental Indication and Application on Immunotherapy. Front Immunol. 2022 Jan 14;12:808964. doi: 10.3389/fimmu.2021.808964. PMID: 35095898; PMCID: PMC8795622.
  12. Iseki Y, Shibutani M, Maeda K, Nagahara H, Fukuoka T, Matsutani S, Kashiwagi S, Tanaka H, Hirakawa K, Ohira M. A new method for evaluating tumor-infiltrating lymphocytes (TILs) in colorectal cancer using hematoxylin and eosin (H-E)-stained tumor sections. PLoS One. 2018 Apr 26;13(4):e0192744. doi: 10.1371/journal.pone.0192744. PMID: 29698402; PMCID: PMC5919485.
  13. Karki S, Pariyar S. Tumor-infiltrating lymphocytes in colorectal carcinoma. J Pathol Nepal. 2021;11(2):1859–63. doi: 10.3126/jpn.v11i2.38227.
  14. Salgado R, Denkert C, Demaria S, Sirtaine N, Klauschen F, Pruneri G, et al.; International TILs Working Group 2014. The evaluation of tumor-infiltrating lymphocytes (TILs) in breast cancer: recommendations by an International TILs Working Group 2014. Ann Oncol. 2015 Feb;26(2):259-71. doi: 10.1093/annonc/mdu450. Epub 2014 Sep 11. PMID: 25214542; PMCID: PMC6267863.
  15. Soraya F, Sandhika W, Wiratama PA. 8-OHdG and Nrf2 Protein are Expressed Consistently in Various T Stages of Invasive Breast Carcinoma. Asian Pac J Cancer Prev. 2025 Jan 1;26(1):301-307. doi: 10.31557/APJCP.2025.26.1.301. PMID: 39874013; PMCID: PMC12082425.
  16. Nguyen LH, Goel A, Chung DC. Pathways of Colorectal Carcinogenesis. Gastroenterology. 2020 Jan;158(2):291-302. doi: 10.1053/j.gastro.2019.08.059. Epub 2019 Oct 14. PMID: 31622622; PMCID: PMC6981255.
  17. Malki A, ElRuz RA, Gupta I, Allouch A, Vranic S, Al Moustafa AE. Molecular Mechanisms of Colon Cancer Progression and Metastasis: Recent Insights and Advancements. Int J Mol Sci. 2020 Dec 24;22(1):130. doi: 10.3390/ijms22010130. PMID: 33374459; PMCID: PMC7794761.
  18. Tsokkou S, Konstantinidis I, Papakonstantinou M, Chatzikomnitsa P, Liampou E, Toutziari E, et al. Sex Differences in Colorectal Cancer: Epidemiology, Risk Factors, and Clinical Outcomes. J Clin Med. 2025 Aug 6;14(15):5539. doi: 10.3390/jcm14155539. PMID: 40807160; PMCID: PMC12347225.
  19. Chen  K, Collins  G, Wang H, Toh JWT. Pathological Features and Prognostication in Colorectal Cancer. Curr Oncol. 2021 Dec 13;28(6):5356-5383. doi: 10.3390/curroncol28060447. PMID: 34940086; PMCID: PMC8700531.
  20. Wang R, Lian J, Wang X, Pang X, Xu B, Tang S, et al. Survival rate of colorectal cancer in China: A systematic review and meta-analysis. Front Oncol. 2023 Mar 3;13:1033154. doi: 10.3389/fonc.2023.1033154. PMID: 36937415; PMCID: PMC10020492.
  21. Tsabit SS, Trianto HF, Pratiwi SE, Hartono H. Clinicopathological Profile of Colorectal Adenocarcinoma in the Anatomical Pathology Laboratory of Dr . Soedarso Hospital Pontianak. Indones J Cancer. 2023;17 (4):292–8. doi: https://doi.org/10.33371/ijoc.v17i4.1004.
  22. Yao N, Li W, Wang J, Chu H, Duan N, Niu X, et al. Prognostic implications of T stage in different pathological types of colorectal cancer: an observational study using SEER population-based data. BMJ Open. 2024 Feb 29;14(2):e076579. doi: 10.1136/bmjopen-2023-076579. PMID: 38423773; PMCID: PMC10910631.
  23. Testa U, Castelli G, Pelosi E. Genetic Alterations of Metastatic Colorectal Cancer. Biomedicines. 2020 Oct 13;8(10):414. doi: 10.3390/biomedicines8100414. PMID: 33066148; PMCID: PMC7601984.
  24. Farhat F, Daulay ER, Chrestella J, Williamson O, Syari RP. Expressions of Nuclear Factor-kappa B and Peroxisome Proliferator-activated Receptor-Gamma Proportional with Clinical Staging of Nasopharyngeal Carcinoma. Open Access Maced J Med Sci. 2021;9(B):1347–51. doi: 10.3889/oamjms.2021.6261.
  25. Berkovich L, Gerber M, Katzav A, Kidron D, Avital S. NF-kappa B expression in resected specimen of colonic cancer is higher compared to its expression in inflammatory bowel diseases and polyps. Sci Rep. 2022 Oct 5;12(1):16645. doi: 10.1038/s41598-022-21078-7.
  26. Bahrami A, Khalaji A, Bahri Najafi M, Sadati S, Raisi A, Abolhassani A, et al. NF-κB pathway and angiogenesis: insights into colorectal cancer development and therapeutic targets. Eur J Med Res. 2024 Dec 19;29(1):610. doi: 10.1186/s40001-024-02168-w. PMID: 39702532; PMCID: PMC11658081.
  27. Ghadyani R, Mozooni Z, Sohbatzadeh Z, Gachkar L, Sepehr Kahrizi, Movafagh A. Expression patterns and clinical significance of MMP-8, MMP-9 and MMP-13 in colorectal cancer. Cell Mol Biol (Noisy-le-grand). 2025 Oct 7;71(9):111-116. doi: 10.14715/cmb/2025.71.9.14. PMID: 41054365.
  28. Shoari A, Ashja Ardalan A, Dimesa AM, Coban MA. Targeting Invasion: The Role of MMP-2 and MMP-9 Inhibition in Colorectal Cancer Therapy. Biomolecules. 2024 Dec 30;15(1):35. doi: 10.3390/biom15010035. PMID: 39858430; PMCID: PMC11762759.
  29. Matsutani S, Shibutani M, Maeda K, Nagahara H, Fukuoka T, Iseki Y, et al. Tumor-infiltrating Immune Cells in H&E-stained Sections of Colorectal Cancer Tissue as a Reasonable Immunological Biomarker. Anticancer Res. 2018 Dec;38(12):6721-6727. doi: 10.21873/anticanres.13041. PMID: 30504382.
  30. Kraja FP, Jurisic VB, Hromić-Jahjefendić A, Rossopoulou N, Katsila T, Mirjacic Martinovic K, et al. Tumor-infiltrating lymphocytes in cancer immunotherapy: from chemotactic recruitment to translational modeling. Front Immunol. 2025 May 22;16:1601773. doi: 10.3389/fimmu.2025.1601773. PMID: 40475782; PMCID: PMC12137109.
  31. Lalle G, Twardowski J, Grinberg-Bleyer Y. NF-κB in Cancer Immunity: Friend or Foe? Cells. 2021 Feb 9;10(2):355. doi: 10.3390/cells10020355. PMID: 33572260; PMCID: PMC7914614.

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Received January 26, 2026.
Accepted March 20, 2026.
© 2026 The Author(s). International Journal of Biomedicine is published by IMRDC.
This is an open access article under the CC BY-NC-ND 4.0 license.