Characterization of a highly invasive and spontaneously metastatic human malignant melanoma cell line

Abstract

Although the incidence of, and deaths due to, malignant melanoma are rising at a rapid rate, few experimental models mimic the highly metastatic properties associated with the pathogenesis of the human disease, making study of the disease difficult. Thus, new human models are required to understand melanoma biology, especially its metastatic properties. Here we describe C8161, a highly invasive and spontaneously metastatic human melanoma cell line, which grows progressively in the subcutis of athymic nude mice with an average doubling time of approximately 6 days. By the time the tumor reaches a diameter of 1 cm, amelanotic metastases in lymph nodes, skin, peritoneal wall, spleen and lungs have formed. By comparing C8161 to variants from other well-characterized human malignant melanomas (A375 and MeWo) with differing metastatic traits, properties presumed to be involved in metastatic propensity were examined. C8161 showed a 2- to 14-fold higher ability to invade reconstituted basement membrane barriers in the MICS and correspondingly high type-IV collagenase mRNA levels and collagenolytic activity, as compared with other melanoma cell lines. Likewise, differential adhesion to immobilized RBM or HUVEC monolayers was observed, but did not correlate to rank orders of malignant properties. Recently, a correlation between surface expression of ICAM-1 and secondary tumor formation by human melanomas has been described in several laboratories. Basal levels of ICAM-1 on C8161, A375 and MeWo human melanomas were compared, but no correlation with metastatic potential was noted. Proto-oncogene expression in C8161 cells was compared with A375P and A375M variants using Northern blot analysis. c-myc expression was 6-fold greater than both A375 variants; c-fos expression was 3.4-fold less than A375P and 1.7-fold less than A375M; c-jun in C8161 cells was 2.5-fold and 2.1-fold greater than expression in A375P and A375M, respectively. Because C8161 is so highly malignant, amenable to experimental manipulation, and its behavior in nude mice mimics the clinical course of malignant melanoma, this cell line will prove valuable for studying properties associated with human melanoma tumor progression.

Reprogramming metastatic melanoma cells to assume a neural crest cell-like phenotype in an embryonic microenvironment

“…The adult human metastatic cutaneous melanoma cell line C8161 and its poorly aggressive isogenic counterpart C81-61 were isolated from an abdominal wall metastasis (35) and maintained as described in ref. 9.…”

Vascular Channel Formation by Human Melanoma Cells in Vivo and in Vitro: Vasculogenic Mimicry

“…Cell lines derived from primary choroidal or ciliary body melanomas or from foci of metastatic uveal melanoma to the liver 16 and normal melanocytes, as described previously, 17 as well as the human cutaneous metastatic melanoma cell line C8161, 18 were maintained in Dulbecco's modified Eagle's medium (DMEM, Life Technologies, Gaithersburg, MD) supplemented with 10% fetal bovine serum (FBS, Gemini Bioproducts, Calabasas, CA). 19 Endothelial cells derived from pulmonary, brain, and dermal microvasculature (HMEC-1), HUVECs, and embryonic chick endothelial cells were maintained in DMEM, 20% FBS, or 1ϫ MITOϩ (Collaborative Biomedical, Bedford, MA).…”

Expression Profiling of Galectin-3-Depleted Melanoma Cells Reveals its Major Role in Melanoma Cell Plasticity and Vasculogenic Mimicry

“…46 Notably, C8161 is a cell line capable of forming spontaneous metastases in mouse models. 47 To gain further insights into the protumorigenic and prometastatic potential of Gal-3, we studied the effect of its silencing on these cells' capacity for invasion, migration, and tube-like formation in vitro. As shown in Figure 5A, Gal-3 silencing in the more aggressive C8161-c9 cells resulted in a threefold inhibition in their invasion capability through Matrigel-coated filters (P ϭ 0.003, NTshRNA versus Gal3shRNA).…”