Genetically-defined novel oral squamous cell carcinoma cell lines for the development of molecular therapies

Abstract

Emerging biological and translational insights from large sequencing efforts underscore the need for genetically-relevant cell lines to study the relationships between genomic alterations of tumors, and therapeutic dependencies. Here, we report a detailed characterization of a novel panel of clinically annotated oral squamous cell carcinoma (OSCC) cell lines, derived from patients with diverse ethnicity and risk habits. Molecular analysis by RNAseq and copy number alterations (CNA) identified that the cell lines harbour CNA that have been previously reported in OSCC, for example focal amplications in 3q, 7p, 8q, 11q, 20q and deletions in 3p, 5q, 8p, 18q. Similarly, our analysis identified the same cohort of frequently mutated genes previously reported in OSCC including TP53, CDKN2A, EPHA2, FAT1, NOTCH1, CASP8 and PIK3CA. Notably, we identified mutations (MLL4, USP9X, ARID2) in cell lines derived from betel quid users that may be associated with this specific risk factor. Gene expression profiles of the ORL lines also aligned with those reported for OSCC. By focusing on those gene expression signatures that are predictive of chemotherapeutic response, we observed that the ORL lines broadly clustered into three groups (cell cycle, xenobiotic metabolism, others). The ORL lines noted to be enriched in cell cycle genes responded preferentially to the CDK1 inhibitor RO3306, by MTT cell viability assay. Overall, our in-depth characterization of clinically annotated ORL lines provides new insight into the molecular alterations synonymous with OSCC, which can facilitate in the identification of biomarkers that can be used to guide diagnosis, prognosis, and treatment of OSCC.

Resistance to dasatinib is associated with the activation of Akt in oral squamous cell carcinoma

“…Of note, inactivating mutations in CDKN2a/p14 ARF have been proposed to be a predictive marker for dasatinib sensitivity in several cancer cell lines of different types including oral cavity lesions. 25 However, in our ORL cell system, we noted from our RNAseq data 27 that no correlation between dasatinib sensitivity and CDKN2a/p14 ARF mutation was found. This difference may be due to the small sample size of oral cancer lines analysed by Garnett et al 25 compared to the cohort size of the current study.…”

“…A panel of 16 ORL oral squamous cell carcinoma lines cell lines established and maintained as previously reported were used in this study. 26,27 Authentication to original tumour tissues and/or blood samples from the corresponding patients was done as described previously. 27 Two cell lines derived from adenocarcinoma of the lung (HCC827 and A549) were purchased from ATCC (VA, USA) and included as controls.…”

“…26,27 Authentication to original tumour tissues and/or blood samples from the corresponding patients was done as described previously. 27 Two cell lines derived from adenocarcinoma of the lung (HCC827 and A549) were purchased from ATCC (VA, USA) and included as controls. These lines have been previously demonstrated to be sensitive (HCC827) and insensitive (A549) to dasatinib.…”

QuickCount ® : a Novel Automated Software for Rapid Cell Detection and Quantification

“…ORL-150, a human oral squamous cell carcinoma cell line was cultured as described previously [9]. The Click-iT 5-ethynyl-2-deoxyuridine (EdU) assay kit (Invitrogen, CA, USA) was used to measure cell proliferation as described previously [9].…”

“…ORL-150, a human oral squamous cell carcinoma cell line was cultured as described previously [9]. The Click-iT 5-ethynyl-2-deoxyuridine (EdU) assay kit (Invitrogen, CA, USA) was used to measure cell proliferation as described previously [9]. Briefly, cells were seeded on cover slips overnight prior to the addition of 10M EdU and cells were incubated between 2 and 6h as optimized previously, to allow the incorporation of EdU.…”