Detection of lymph node metastasis in colon cancer by ectopically expressed fibroblast markers FOXQ1 and THBS2

Haytham Ali; Manar AbdelMageed; Lina Olsson; Gudrun Lindmark; Marie-Louise Hammarström; Sten Hammarström; Basel Sitohy

doi:10.3389/fonc.2023.1297324

Detection of lymph node metastasis in colon cancer by ectopically expressed fibroblast markers FOXQ1 and THBS2

Front Oncol. 2023 Dec 14:13:1297324. doi: 10.3389/fonc.2023.1297324. eCollection 2023.

Authors

Haytham Ali^{1

2

3}, Manar AbdelMageed^{1

2

4}, Lina Olsson¹, Gudrun Lindmark⁵, Marie-Louise Hammarström¹, Sten Hammarström¹, Basel Sitohy^{1

2}

Affiliations

¹ Department of Clinical Microbiology, Umeå University, Umeå, Sweden.
² Department of Radiation Sciences, Oncology, Umeå University, Umeå, Sweden.
³ Department of Animal and Veterinary Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Muscat, Oman.
⁴ Department of Pathology, Faculty of Veterinary Medicine, Zagazig University, Zagazig, Egypt.
⁵ Institution of Clinical Sciences, Lund University, Lund, Sweden.

Abstract

Introduction: Approximately 25% of colon cancer (CC) patients having curative surgery will relapse. Therefore, it is crucial to identify patients with increased recurrence risk to offer them adjuvant chemotherapy. Three markers with prominent expression in fibroblasts: forkhead box Q1 (FOXQ1), matrix metalloproteinase-11 (MMP11), and thrombospondin-2 (THBS2), and the fibroblast expressed chemokine CXCL12 were selected for studies because of the critical role of fibroblasts in the microenvironment of the tumor.

Methods: The expression levels of the biomarkers were assessed in primary CC tumors, lymph nodes of CC patients and controls, and CC cell lines at mRNA and protein levels by real-time qRT-PCR and immunohistochemistry, respectively.

Results: FOXQ1, MMP11, and THBS2 mRNAs were expressed at significantly higher levels in primary tumors compared to normal colon (P=0.002, P<0.0001, and P<0.0001, respectively). In contrast, CXCL12 mRNA levels were higher in normal colon tissue. FOXQ1, MMP11, and THBS2 levels were also expressed at significantly higher levels in metastasis-positive lymph nodes compared to both metastasis-negative- and control nodes (P<0.0001/P=0.002, P<0.0001/P<0.0001, and P<0.0001/P<0.0001, respectively). Immuno-morphometry revealed that 30-40% of the tumor cells expressed FOXQ1, MMP11, and THBS2. FOXQ1 and THBS2 were barely detected in normal colon epithelium (P<0.0001), while MMP11 was expressed in normal colon epithelium at high levels.

Discussion: We conclude that CC tumor cells show ectopic expression of FOXQ1 and THBS2 possibly making these tumor cells independent of fibroblast cell support. The high expression levels of these two biomarkers in metastatic lymph nodes suggest that they are potential indicators of patients at risk for recurrence.

Keywords: CXCL12; FOXQ1; MMP11; THBS2; colon cancer; fibroblasts; immunohistochemistry; qRT-PCR.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was financially supported by grants from the Swedish Research Council-Medicine and Health (BS) and the Swedish Research Council-Natural and Engineering Sciences (M-LH) Grant numbers: 2010-05669 and 2013-04522, the Medical Faculty of Umeå University (BS and M-LH), the County Council of Västerbotten (BS), Kempe Foundation (BS) and the Lions Cancer Research Fund (BS).