# ITIH2 in colorectal cancer metastasis: Weighted Gene Co-expression Network Analysis-guided functional validation

## TL;DR

This study identifies ITIH2 as a key gene linked to colorectal cancer liver metastases, suggesting it could be a new biomarker and treatment target.

## Contribution

The study functionally validates ITIH2 as a novel hub gene associated with CRC liver metastasis using bioinformatics and experimental approaches.

## Key findings

- ITIH2 expression is significantly higher in CRC liver metastases compared to primary tumors.
- ITIH2 overexpression increases CRC cell proliferation, motility, and invasion, while knockdown inhibits these behaviors.
- ITIH2 is correlated with poor clinical outcomes and has independent prognostic significance for overall survival.

## Abstract

One of the main causes of death from colorectal cancer (CRC) is liver metastases; yet, little is known about the genetic factors that influence the development of these metastases. Given their proven involvement in cancer spread, we anticipated that major differentially expressed genes (DEGs) between primary and metastatic CRC could identify important molecular players in liver metastasis, with an emphasis on extracellular matrix and proteolytic processes. The purpose of this study was to use integrative bioinformatics and experimental validation to identify and functionally describe hub genes linked to CRC liver metastasis.

The differentially expressed genes (DEGs) between primary (CRC and CRC liver metastases are investigated using two microarray datasets (GSE14297 and GSE6988). Weighted Gene Co-expression Network Analysis (WGCNA) was used to generate co-expression networks and identify functional modules associated with metastatic progression. Topological research revealed that ITIH2 is an essential hub gene. To investigate its functional role, we employed lentiviral transduction to generate stable CRC cell lines with ITIH2 overexpression and knockdown. For in vivo validation, liver metastasis models were created in BALB/c mice using engineered cell lines. To further explore clinical relevance, ITIH2 expression in tissue microarrays from patients with primary and metastatic CRC was examined immunohistochemically.

When compared to primary tumors, ITIH2 expression was noticeably higher in CRC liver metastases. In contrast to ITIH2 knockdown, which inhibited these malignant behaviors, functional studies showed that ITIH2 overexpression increased CRC cell proliferation, motility, and invasion. These results were supported in vivo, where ITIH2 expression increased the likelihood of tumor growth and metastasis.

ITIH2 plays a functional role in metastatic behavior and is markedly elevated in liver metastases of CRC. We conclude that ITIH2 may be a novel prognostic biomarker and a potential therapeutic target in patients with CRC who have liver metastases due to its high correlation with poor clinical outcomes and independent prognostic significance for overall survival.