Establishment and characterization of novel autologous pair cell lines from two Indian non‑habitual tongue carcinoma patients.

Abstract

Oral tongue squamous cell carcinoma (OTSCC) is one of the major causes of fatality in India due to very high percentage of patients with habits of smoking and chewing tobacco and associated products. Being highly heterogeneous in nature, every patient poses a different challenge clinically. To understand disease progression in an improved way, knowledge of cross‑talk between tumor stroma and the tumor cells becomes indispensable. Patient‑derived invitro cell line models are helpful to understand the complexity of diseases. However, they have very low efficiency of establishment from the tumor samples, particularly the cancer‑associated fibroblasts (CAFs). In the present study, two novel autologous pairs were immortalized spontaneously from non‑habitual, HPV‑positive patients, who presented with OTSCC. The epithelial and fibroblast cell lines had typical polygonal and spindle‑shaped morphology, respectively. Positive staining with epithelial specific Pan‑cytokeratin (PanCK) and fibroblast specific protein (FSP‑1) further confirmed their epithelial and fibroblast origin. Unique Short Tandem Repeat (STR) profile of the cultures confirmed their novelty, while the similarity of the STR profiles between the epithelial and fibroblast cells from the same patient, confirmed their autologous nature. DNA analysis revealed aneuploidy of the established cultures. An increase in the tumorigenic potential of the established epithelial cultures upon treatment with CAF‑conditioned medium proved the 'CAF‑ness' of the established fibroblast cells. The established cultures are the first of their kind which would serve as a useful platform in understanding the tumor‑stroma cross‑talk in tongue cancer progression.