PO-342 Establishment and molecular profile of a new cervical cancer cell line from brazilian patient biopsy

Introduction In vitro model using continuous tumour cell lines is very common in the screening of new drugs in the treatment of cancer. However, the low number of commercially available cervical cancer cell lines (CCCL) poses a challenge to generate an effective and representative means of screening. There is thus a need to establish new CCCL to improve this process. Therefore, we aimed to establish and to characterise a CCCL from a Brazilian patient. Material and methods Following patient‘s consent (Ethics Committee HCB-985/2015), a biopsy sample was taken, fragmented and subjected to enzymatic digestion. The material was then cultured in a flask with keratinocyte medium and when cells reached sixth passage, they were submitted to immunocytochemistry to characterise its phenotype. To investigate the HPV status, PCR for GP5+/GP6 +sequences and the HPV test using COBAS were carried out. Short tandem repeat (STR) analysis from cell line and patient´s blood was done to confirm the patient origin. Following DNA isolation of cell line and patient‘s blood, whole-exome sequencing (WES) and copy number variation (CNV) assays were performed. Moreover, functional assays are being performed to evaluate its tumorigenic potential protein profile. Results and discussions Overall, 35 tissues were collected and attempted to be established in vitro . Only one (2,9%) of the cultures survived for more than 3 months. The successful cell line, named HCB-514, was derived from a stage IIB squamous cell carcinoma of a 30 years old patient, with no previous treatment. It continued to grow after freeze-thaw cycles and showed positive expression for AE1/AE3 cytokeratin, confirming its epithelial phenotype. The STR analysis of HCB-514 compared with the patient’s blood confirmed its lineage. The HCB-514 was HPV 16 positive. The WES identified 93 non-silent mutations, among them 11 (HPS-3, IRX2, XPO5, CBWD1, TGFBR1, CUBN, TRHDE, BRCA2, THBS1, TGM5, CCDC22) have potentially pathogenic. CNV revealed 44 somatic alteration, including 30 regions with gains (NF1, CCND1, MYC, TP53, DCC, MAP2K4, AURKA, IGF1R, PIK3CA, GAB2, NKX2-1, BCL2L2, C8orf4, BCL2L1, ZNF217, EGFR, NCOA3, SKP2, FADD, ORAOV1, EEF1A2, REL, TERT, AKT3, PRKCI, MAPK7, DCUN1D1, BIRC2, YAP1, MYCN) and 14 losses (FHIT, GPC5, IRS2, BRCA2, RB1, PDGFRA, CDK6, SHH, APC, FOXO1, MET, MITF, KDR, KIT). Conclusion The characterisation of a new cervical cancer cell line, HCB-514, will constitute an important in vitro tool for further biological and therapeutic studies of cervical cancer.