Developing a Stepwise Induction Protocol for Cisplatin-Resistant Human Cervical Cancer Cells

Abstract

Cisplatin, a platinum-based DNA crosslinking chemotherapeutic, remains the cornerstone of first-line chemoradiation therapy for advanced cervical cancer. However, the clinical efficacy of cisplatin is frequently compromised by the emergence of acquired resistance, which represents a principal cause of treatment failure and disease recurrence. Traditional protocols for generating resistant cervical cancer cell lines vary widely in dose-escalation schedules, passage criteria, timing, and endpoints, often requiring months to yield inconsistent results. To overcome these limitations, this study establishes a structured, stepwise protocol for developing cisplatin-resistant cervical cancer cell lines using HeLa (HPV18+) and SiHa (HPV16+) models. We hypothesised that a predefined dosing and monitoring scheme would shorten induction time while improving the reproducibility of resistance generation. Using this approach, we successfully generated cisplatinresistant sublines (H1CR and S1CR) that exhibited rightward shifts in cisplatin IC50 (1.28-fold in HeLa and 1.80-fold in SiHa) and stable resistant phenotypes after drug withdrawal and cryopreservation. The present work therefore focuses on methodological standardisation of cisplatin resistance induction rather than exhaustive molecular characterisation, providing a clear starting point for future mechanistic and translational studies.