Orthotopic non-metastatic and metastatic oral cancer mouse models

Abstract

SUMMARY Oral cancer is characterized by high morbidity and mortality with a predisposition to metastasize to different tissues, including lung, liver, and bone. Despite progress in the understanding of mutational profiles and deregulated pathways in oral cancer, patient survival has not significantly improved over the past decades. Therefore, there is a need to establish in vivo models that recapitulate human oral cancer metastasis to evaluate therapeutic potential of novel drugs. Here we report orthotopic tongue cancer nude mouse models to study oral cancer growth and metastasis using human metastatic (UMSCC2) and non-metastatic (CAL27) cell lines, respectively. Transduction of these cell lines with lentivirus expressing red fluorescent protein (DsRed) followed by injection into tongues of immunodeficient mice generated orthotopic tongue tumors that could be monitored for growth and metastasis by fluorescence measurement with an in vivo Imaging System (IVIS 200). The growth rates of CAL27-DsRed induced tumors were higher than UMSCC2-DsRed tumors after day 15, while UMSCC2-DsRed tumors revealed metastasis beginning on day 21. Importantly, UMSCC2 tumors metastasized to a number of tissues including the submandibular gland, lung, kidney, liver, and bone. Further, immunohistochemical analyses of tongue tumors induced by CAL27 and UMSCC2 cells revealed elevated expression of components of protumorigenic pathways deregulated in human cancers, including Cyclin D1, PCNA, Ki-67, LSD1, LOXL2, MT-MMP1, DPAGT1, E-cadherin, OCT4A, and H3K4me1/2. These orthotopic mouse models are likely to be useful tools for gaining insights into the activity and mechanisms of novel oral cancer drug candidates.

Determination of cell uptake pathways for tumor inhibitor lysyl oxidase propeptide

“…rLOX‐PP has been demonstrated to inhibit models of prostate and breast cancer growth in vitro and in vivo, and we have recently developed an orthotopic model of human oral cancer growth and metastasis in mice (Bais et al., 2015a). Here we first wished to determine whether ectopic expression of rLOX‐PP could inhibit the growth and metastasis of UMSCC2 metastatic oral cancer cells implanted in mouse tongues.…”

Role for the Aryl Hydrocarbon Receptor and Diverse Ligands in Oral Squamous Cell Carcinoma Migration and Tumorigenesis

“…We recently described a novel orthotopic xenograft model for human OSCC [32]. Here, this model was used to further explore the role of the AHR in OSCC and to begin to assess if AHR inhibition represents a viable, targeted approach to OSCC therapy.…”

“…CAL27 cells (3 x 10 5 ) were injected into the tongues of six-week old female NCr nude mice (CrTac:NCr- Foxn1 nu , Taconic, Hudson, NY) as we previously described [32]. Mice began a daily lavage schedule with 50 μl sesame oil alone or CB7993113 dissolved in sesame oil (50 mg/kg/day in 50 μl) 24 hours before tongue injections.…”

“…To address the possible contribution of environmental and microbial AHR ligands to OSCC, several classes of AHR ligands were used including a genotoxic PAH (B(a)P), a non-genotoxic but persistent halogenated hydrocarbon (TCDD), PAH-containing 2.5 μm airborne particles (PM 2.5 ), and products from Pseudomonas aeruginosa and Porphyromonas gingivalis , the latter a primary etiological agent associated with chronic periodontitis. To assess the feasibility of targeting the AHR signaling pathway as a novel OSCC treatment, we used two OSCC cell lines, a unique OSCC orthotopic xenograft model [32], a novel AHR inhibitor (CB7993113) [33], and molecular knockdown techniques.…”

Lysyl Oxidase Isoforms and Potential Therapeutic Opportunities for Fibrosis and Cancer

“…It is widely over-expressed in cancers, including metastatic cancers. For example, high levels of LOXL2 occur in colon cancer associated fibroblasts 128 , skin cancer 129 , oral cancer 130 , skin and head and neck squamous cell carcinoma 101 , liver metastasis 103 , cholangiocarcinoma 131 , breast cancer 76, 92, 132, 133 , and gastric cancer metastasis 102 . Attenuation of LOXL2 expression was shown to inhibit metastasis or the invasive cell phenotypes in most of these studies.…”