CHARACTERIZATION OF A NEW CISPLATIN-RESISTANT CELL LINE DERIVED FROM NON-SMALL LUNG CANCER (NSCLC) CELL LINE NCI-H125

BECHIS, ANDRES; BRITEZ NEIRA, DIEGO JAVIER; URTREGER, ALEJANDRO JORGE; TODARO, LAURA BEATRIZ

Mar del Plata, Buenos Aires

Congreso; REUNIÓN DE SOCIEDADES DE BIOCIENCIAS 2023; 2023

Sociedad Argentina de Investigación Clínica

Cisplatin (CDDP) is a non-small cell lung cancer (NSCLC) standard therapy. The frequent resistance acquisition to this treatment would be associated with the presence of cancer stem cells (CSC). The generation of CDDP-resistant cell lines is an important tool to study the mechanisms involved in this characteristic and the parallelism with what happened with the patients. In order to generate the H125 CDDP-resistant cell line (H125cpr), we treated the parental H125 cell line with CDDP 1μM in 72h weekly pulses during 12 weeks. After that, we escalate the treatment dose to 2μM in 72h weekly pulses for 20 more weeks. After 32 treatment weeks, H125cpr showed a IC50 of 6μM, while H125ct (control cell line without treatment) presented a IC50 of 1.5μM. To corroborate CDDP resistance, we evaluate the distribution of the cells in the different phases of their cell cycle by flow cytometry and propidium iodide dye. We observed that CDDP treatment produced an arrest in G2-M phase in H125ct, while this treatment had no effect in this parameter in H125cpr. Besides, we observed that H125cpr has a lower migratory capability than H125ct. In the other hand, we observed that H125cpr has a bigger nucleus surface than H125ct. Besides, H125cpr presents more cells with cytoplasmic chromatin fragments (CCF) and a lower duplication rate than H125ct. These results indicated that H125cpr would have a more senescent/stem like phenotype than H125ct. So, we performed an oncosphere formation assay and we corroborated that H125cpr oncospheres showed higher growth rate than H125ct ones. Taking all together, we hypothesize that H125cpr has higher proportion cells with a phenotype more stem-like than H125ct. More experiments are needed to elucidate the mechanisms implied in the CDDP-resistance.