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

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 (H 1 CR and S 1 CR) 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.