Insights into the characteristics of primary radioresistant cervical cancer using single-cell transcriptomics

Abstract

Radioresistance is a major cause of radiotherapy failure among patients with cervical cancer (CC), the fourth most common cause of cancer mortality in women worldwide. Traditional CC cell lines lose intra-tumoral heterogeneity, posing a challenge for radioresistance research. Meanwhile, conditional reprogramming (CR) maintains intra-tumoral heterogeneity and complexity, as well as the genomic and clinical characteristics of original cells and tissues. Three radioresistant and two radiosensitive primary CC cell lines were developed under CR conditions from patient specimens, and their characteristics were verified via immunofluorescence, growth kinetics, clone forming assay, xenografting, and immunohistochemistry. The CR cell lines had homogenous characteristics with original tumor tissues and maintained radiosensitivity in vitro and in vivo, while also maintaining intra-tumoral heterogeneity according to single-cell RNA sequencing analysis. Upon further investigation, 20.83% of cells in radioresistant CR cell lines aggregated in the G2/M cell cycle phase, which is sensitive to radiation, compared to 38.1% of cells in radiosensitive CR cell lines. This study established three radioresistant and two radiosensitive CC cell lines through CR, which will benefit further research investigating radiosensitivity in CC. Our present study may provide an ideal model for research on development of radioresistance and potential therapeutic targets in CC.

Conditional Cell Reprogramming and Air–Liquid Interface Modeling Life Cycle of Oncogenic Viruses (HPV and EBV) in Epithelial Cells and Virus-Associated Human Carcinomas

“…Recently, CR technology was used to establish three radioresistant and two radiosensitive cell lines from cervical cancer patients’ specimens and verified their molecular characteristics and biological changes including growth kinetics, immunofluorescence, clone forming assay xenografting, and immunohistochemistry vitro and in vivo [ 58 ]. The results of single-cell RNA (sc-RNA) sequencing analysis showed that 38.1% of cells in radiosensitive CR cell lines aggregated in the G2/M cell cycle phase which is sensitive to radiation compared to 20.83% of cells in radioresistant CR cell lines.…”