B16/F10 tumors in aged 3D collagen in vitro simulate tumor growth and gene expression in aged mice in vivo

Abstract

Although the incidence of cancer rises with age, tumor growth is often slowed in older hosts. The B16/F10 melanoma cell line is commonly used in murine models of age-related tumor growth suppression. We wished to determine if the growth pattern and gene expression of B16/10 tumors grown in aged mice could be simulated in 3D collagen matrices derived from aged mice. Outcome measures were tumor size in vitro and gene expression of the key growth regulatory molecules: growth hormone receptor (GHR), IL-10Rβ, IL-4Rα, and IL-6. B16/F10 tumors were grown in 20–25-mo-old C57/BL6 male mice. Tumor sizes ranged from 30 to 4,910 mg in vivo. Tumors from a subset of mice were removed after euthanasia, and equivalent amounts of each tumor were placed in aged 3D collagen and grown for 5 d. Tumor sizes in aged 3D collagen correlated highly with their original tumor size in vivo. Gene expression changes noted in vivo were also maintained during tumor growth in aged 3D collagen in vitro. The relative expression of GHR was increased, IL-10Rβ was unchanged, and IL-4Rα and IL-6 were decreased in the larger tumors relative to the smaller tumors in vitro, in a pattern similar to that noted in vivo. We propose that 3D matrices from aged mice provide an in vitro model of tumor growth that correlates highly with tumor size and expression of key regulatory molecules in vivo.

Growth hormone is increased in the lungs and enhances experimental lung metastasis of melanoma in DJ-1 KO mice

“…Moreover, GHRs have been demonstrated in human and murine melanoma cells [20, 23]. We therefore examined the effects of GH on cultured melanoma cells.…”

Hyaluronan in aged collagen matrix increases prostate epithelial cell proliferation

“…In this context, we and others have established that 3D gels polymerized from collagen extracted from young and aged animals induce differential cellular responses that reflect tissue aging (Jiang et al 2000; Bartling et al 2009; Damodarasamy et al 2010). Moreover, aged collagenous ECM in vitro provides an environment that can simulate pathologic responses in vivo (Bentov et al 2013). …”

“…Collagenous ECM extracted from tail tendons and polymerized into 3D gels provides an ideal model to study the effect of specific ECM components on cell behaviors (Bartling et al 2009; Leventhal et al 2009). Three-dimensional gels made from collagen extracted from aged animals are highly useful tools to study the effects of aged ECM on cell behaviors in vitro, which are relevant to cell-ECM interactions in vivo (Bentov et al 2013). In this context, we found that, compared to young collagen gels, aged collagen gels elicited significantly greater proliferation of three distinct prostate epithelial cell lines: PEC, BPH-1, and LNCaP.…”

“…As we and others have previously shown, when compared to gels made from young collagen extracts (“young collagen gels”), gels made from aged collagen extracts (“aged collagen gels”) retain many of the characteristics of native, aged tissues, which include decreased collagen density, greater disorganization and heterogeneity of collagen fibers, and increased levels of advanced glycation end-products (Jiang et al 2000; Wu et al 2005; Bartling et al 2009; Damodarasamy et al 2010). The phenomenon that aged collagen gels made in vitro retain, in some respects, the characteristics of collagenous ECM in native aged tissues in vivo makes these gels useful for in vitro studies of age-associated changes in cell-ECM interactions (Bentov et al 2013). …”