NCTC clone 2472

Characterization of Commercially Available Murine Fibrosarcoma NCTC-2472 Cells

Cancer-induced bone pain (CIBP) is a severe neuropathic pain caused by tumor metastasis to bone, significantly impacting patients' quality of life. Current treatments focus on symptom management, but novel therapeutics are needed. The NCTC-2472 murine fibrosarcoma cell line is commonly used in preclinical CIBP models. This study characterized NCTC-2472 cells both in vitro and in vivo to assess their viability and suitability for evaluating new anti-cancer and analgesic compounds.

NCTC-2472 fibrosarcoma cells did not display prominent morphological markers indicative of apoptosis in regular culture. Throughout continued cell passaging, NCTC-2472 cells exhibited typical cellular morphology associated with fibroblast type cells that include branched cytoplasmic extensions and a general spindle shape body pattern with bipolar presentation (Fig 1a). There was no observed shrinking of the nucleus, no cellular blebbing, minimal presence of cellular debris, and no loss of adherence in low passage numbers (Fig 1b), moderate passage numbers (Fig 1c), or in high passage numbers (Fig 1d) amongst any cells. To determine the self-renewal capacity of NCTC-2472 fibroblasts, we measured and calculated the proliferative ability of the cells via CPDL measurements (Fig 1e). A consistently linear increase in the rate of cell proliferation was observed and indicated that despite prolonged passaging, there was no impact on cellular proliferation.

Fig. 1. Culture of NCTC-2472 fibrosarcoma cells and identification of cumulative population doubling level (CPDL) (Ortiz YT, Shamir LG, et al., 2024).

LPA Signaling Contributed to Hypersensitivity Produced by Bone Cancer

Bone cancer pain is a severe condition affecting over 60% of patients, with current treatments often being insufficient. The mechanisms involve interactions between cancer cells and nociceptive neurons, with autotaxin (ATX) and lysophosphatidic acid (LPA) playing key roles in cancer progression and pain signaling. This study investigates the role of ATX–LPA–LPAR signaling in cancer pain using in vitro and in vivo methods.

Consistent with prior studies mechanical and heat hypersensitivity developed within 10 days after implanting mouse NCTC 2472 fibrosarcoma cells into the calcaneus bone of male and female mice, with no sex differences. Implanting nonmalignant NCTC 929 fibroblasts caused no tumors or hypersensitivity in naive mice. Tumor-bearing (TB) mice showed elevated serum LPA levels (Fig. 2A), specifically 18:1-, 18:2-, and 20:4-LPA (Fig. 2B), while 16:0- and 22:6-LPA remained unchanged. LPA levels were not measured in rare non-hypersensitive mice. In vitro, 18:1-LPA (10 µM, 24h) promoted fibrosarcoma cell proliferation (Fig. 2C), consistent with prior protocols. Given elevated LPA and its LPAR1,3,5 targeting in DRG, LPAR antagonist H2L5765834 (5 mg/kg, intraplantar) reduced mechanical and heat hypersensitivity in TB mice versus vehicle (Figs. 2D, E). Lower doses (1 mg/kg) or contralateral administration had no effect, indicating localized action.

Fig. 2. Lysophosphatidic acid signaling contributed to hypersensitivity produced by bone cancer (Iryna A K, Sergey G K, et al., 2023).