Combined epithelial marker analysis of tumour budding in stage II colorectal cancer

Abstract

Tumour budding predicts survival of stage II colorectal cancer (CRC) and has been suggested to be associated with epithelial‐to‐mesenchymal transition (EMT). However, the underlying molecular changes of tumour budding remain poorly understood. Here, we performed multiplex immunohistochemistry (mIHC) to phenotypically profile tumours using known EMT‐associated markers: E‐cadherin (adherence junctions), integrin β4 (ITGB4; basement membrane), ZO‐1 (tight junctions), and pan‐cytokeratin. A subpopulation of patients showed high ITGB4 expression in tumour buds, and this coincided with a switch of ITGB4 localisation from the basal membrane of intact epithelium to the cytoplasm of budding cells. Digital image analysis demonstrated that tumour budding with high ITGB4 expression in tissue microarray (TMA) cores correlated with tumour budding assessed from haematoxylin and eosin (H&E) whole sections and independently predicted poor disease‐specific survival in two independent stage II CRC cohorts (hazard ratio [HR] = 4.50 (95% confidence interval [CI] = 1.50–13.5), n = 232; HR = 3.52 (95% CI = 1.30–9.53), n = 72). Furthermore, digitally obtained ITGB4‐high bud count in random TMA cores was better associated with survival outcome than visual tumour bud count in corresponding H&E‐stained samples. In summary, the mIHC‐based phenotypic profiling of human tumour tissue shows strong potential for the molecular characterisation of tumour biology and for the discovery of novel prognostic biomarkers.

Tumour budding, poorly differentiated clusters, and T-cell response in colorectal cancer

“…Tumour budding is one of the initial steps of cancer invasion and metastasis, as budding cells migrate through the extracellular matrix, invade lymphovascular structures, and form metastatic tumour colonies in lymph nodes and distant organs 34 , 35 , 36 . During this process called epithelial mesenchymal transition (EMT), activation of WNT/CTNNB1 signalling occurs in tumour cells, which lose expression of epithelial markers such as CDH1 expression and instead express genes more commonly associated with mesenchymal cells, such as CDH2 (cadherin 2, N-cadherin) and VIM (vimentin) [ 2 , 37 , 38 ].…”

Squalene Epoxidase Correlates E-Cadherin Expression and Overall Survival in Colorectal Cancer Patients: The Impact on Prognosis and Correlation to Clinicopathologic Features

“…In the present study, we observed that there was a significant correlation between a high expression level of SE and the tumor budding status and invasion depth in the CRC. Tumor budding has been suggested to be associated with EMT, as evidenced by decreased or aberrant expression of E-cadherin [39,40]. The SE and E-cadherin protein expression pattern in the tumor budding of the CRC was determined by immunohistochemical analysis.…”

Fibroblast as a critical stromal cell type determining prognosis in prostate cancer

“…The threshold for PanEpi channel was set manually to exclude all of the stromal area based on visual inspection. The PanEpi detection included staining and detection of two different anti‐pan‐cytokeratin antibody clones (AE1/3 and C‐11) and anti‐E‐cadherin antibody (clone 36) for optimal epithelium coverage . Stromal cells were classified as fibroblasts (VIM‐pos, aSMA‐neg), myofibroblasts (VIM‐pos, aSMA‐pos), or smooth muscle (aSMA‐pos, VIM‐neg) (Figure ; Figure S1).…”