# A Novel Serum-Based Bioassay for Quantification of Cancer-Associated Transformation Activity: A Case–Control and Animal Study

## TL;DR

A new blood test detects ovarian cancer by measuring how cancer-related factors in the blood cause cells to grow abnormally.

## Contribution

A novel serum-based bioassay using TY cells to detect cancer-associated transformation activity is introduced.

## Key findings

- TY cells effectively distinguished cancer sera from controls with high statistical significance.
- The AIG-promoting activity in sera correlated strongly with tumor burden in a mouse model.
- HGF/c-MET and IGF/IGF-1R signaling pathways mediate the observed AIG-promoting activity.

## Abstract

Background/Objectives: The detection of ovarian cancer remains challenging due to the lack of reliable serum biomarkers that reflect malignant transformation rather than mere tumor presence. We developed a novel biotest using an immortalized human fallopian tube epithelial cell line (TY), which exhibits anchorage-independent growth (AIG) in response to cancer-associated serum factors. Methods: Sera from ovarian and breast cancer patients, non-cancer controls, and ID8 ovarian cancer-bearing mice were tested for AIG-promoting activity in TY cells. Results: TY cells (passage 96) effectively distinguished cancer sera from controls (68.50 ± 2.12 vs. 17.50 ± 3.54 colonies, p < 0.01) and correlated with serum CA125 levels (r = 0.73, p = 0.03) in ovarian cancer patients. Receiver operating characteristic (ROC) analysis showed high diagnostic accuracy (AUC = 0.85, cutoff: 23.75 colonies). The AIG-promoting activity was mediated by HGF/c-MET and IGF/IGF-1R signaling, as inhibition of these pathways reduced phosphorylation and AIG. In an ID8 mouse ovarian cancer model, TY-AIG colonies strongly correlated with tumor burden (r = 0.95, p < 0.01). Conclusions: Our findings demonstrate that the TY cell-based AIG assay is a sensitive and specific biotest for detecting ovarian cancer and potentially other malignancies, leveraging the fundamental hallmark of malignant transformation.