Characterization of Thyroid Cancer Cell Lines in Murine Orthotopic and Intracardiac Metastasis Models

Abstract

Thyroid cancer incidence has been increasing over time, and it is estimated that ~1950 advanced thyroid cancer patients will die of their disease in 2015. To combat this disease, an enhanced understanding of thyroid cancer development and progression as well as the development of efficacious, targeted therapies are needed. In vitro and in vivo studies utilizing thyroid cancer cell lines and animal models are critically important to these research efforts. In this report, we detail our studies with a panel of authenticated human anaplastic and papillary thyroid cancer (ATC and PTC) cell lines engineered to express firefly luciferase in two in vivo murine cancer models- an orthotopic thyroid cancer model as well as an intracardiac injection metastasis model. In these models, primary tumor growth in the orthotopic model and the establishment and growth of metastases in the intracardiac injection model are followed in vivo using an IVIS imaging system. In the orthotopic model, the ATC cell lines 8505C and T238 and the PTC cell lines K1/GLAG-66 and BCPAP had take rates >90% with final tumor volumes ranging 84–214 mm3 over 4–5 weeks. In the intracardiac model, metastasis establishment was successful in the ATC cell lines HTh74, HTh7, 8505C, THJ-16T, and Cal62 with take rates ≥70%. Only one of the PTC cell lines tested (BCPAP) was successful in the intracardiac model with a take rate of 30%. These data will be beneficial to inform the choice of cell line and model system for the design of future thyroid cancer studies.

Establishment and Characterization of Four Novel Thyroid Cancer Cell Lines and PDX Models Expressing the RET/PTC1 Rearrangement, BRAFV600E, or RASQ61R as Drivers

“…The CUTC60 cells readily formed orthotopic and flank tumors in nude mice, demonstrating lung metastases in the orthotopic model. Interestingly, the CUTC48 cells did not form tumors in any mouse model that we tested, including the NRG mice, which is consistent with the take rate we and others have observed with the RET/PTC1-expressing TPC1 cells (34,45,46). Our previous studies have indicated that thyroid cancer cell lines with the highest take rate harbor mutations in BRAF V600E (34), thus indicating that the RET/PTC1-expressing cells may exhibit an overall less aggressive phenotype.…”

“…All animal studies were conducted in accordance with the animal protocol procedures approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Colorado Denver (Aurora, CO). Athymic nude mice [Hsd:Athymic Nude-Foxn1 nu (069) from Envigo or in-house breeding for the CUTC60 and CUTC61 cells], SCID/Beige mice (C.B-17/IcrHsd-Prkdc scid Lyst bg-J from Envigo), or NRG mice [NOD.Cg-Rag1 tm1Mom Il2rg tm1Wjl /SzJ (#007799) from The Jackson Laboratory] were anesthetized with tribromoethanol (250 mg/kg), and the indicated thyroid cancer cells (5 Â 10 5 in a 5 mL cell suspension) were injected into the right thyroid gland with the aid of a dissecting microscope (Nikon SMZ645), and the skin closed with staples, as described previously (33)(34)(35). Growth was monitored for the indicated days.…”

FAK Expression, Not Kinase Activity, Is a Key Mediator of Thyroid Tumorigenesis and Protumorigenic Processes

“…We next evaluated the role of FAK kinase activity versus expression in tumor establishment and growth using the orthotopic thyroid cancer model (9,34–38). To assess the role of FAK kinase activity, BCPAP or 8505C cells engineered to stably express a luciferase-IRES-GFP plasmid were injected into the right thyroid gland of athymic nude mice, as previously described (9).…”

Preclinical Imaging for the Study of Mouse Models of Thyroid Cancer

“…While the xenograft models are quite straightforward in that the cells grown and collected are injected into the subcutaneous tissues of immunocompromised mice, the orthotopic model is more complex to establish, since it requires a microsurgical procedure to directly place the cells into one or both the thyroid lobes [ 32 , 33 , 34 ]. Nonetheless, since the development of such a model [ 35 ], orthotopic implantation has been widely adopted to study the genetic and bio-molecular patterns of thyroid carcinogenesis [ 9 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 ].…”

“…Recently, our group developed a high-frequency ultrasound (HFUS) guided procedure for the orthotopic implantation of FTC cells, demonstrating its feasibility and comparing tumor growth curves with the standard surgical procedure, showing interesting results (see below) [ 34 ]. In any case, the main limitations of such in vivo models are the use of mice with various degrees of immunodeficiency, which precludes studying the interactions between the cancer cells and the host immune system [ 42 ] and hence the ability to predict the efficacy of clinical therapeutics.…”