The aim of this study was to evaluate the anticancer and antitumor activities of ceramide analog 315 in nude mice. Nude mice (n=10) were injected bilaterally with 5×10 MDA-MB-231 cells on each side. Tumors were allowed to form for 2 weeks. The mice were then divided into two groups (n=5 in each group). The control group mice were injected with 25 μl of dimethyl sulfoxide and the treatment group mice were injected with 10 mg/kg of analog 315 (in dimethyl sulfoxide, 25 μl volume) every day for a period of 3 weeks. Animal weights and tumors were measured every week for 3 weeks. At the end of the experimental period, control animals had retained excess fluid, and showed larger tumor sizes compared with the treated group (2.95 vs. 1.67 g). A 45% reduction in tumor size and 80% decrease in tumor volume were observed in the treatment group. There was a significant increase in the weights of liver (10%) and spleen (19%) between the control and treated animals. Hematoxylin and Eosin staining of MDA-MB-231 tumor sections revealed more acellular necrotic regions in tumors from the treatment groups compared with the ones from the control group. Ki67, a proliferation marker was higher in number in control tumor section (71.8±12.8) compared to the treatment tumor section (37.4±10.4) (P<0.001). Photomicrographs showed metastatic tumor burden in kidney, lungs, and spleen collected from the control group mice bearing MDA-MB-231 tumors. Treatment group mice showed normal microscopic tissue architecture. Overall, our study showed tumor growth inhibition and antimetastatic effects for the novel ceramide analog 315.

E‐cigarette usage is growing, especially among adolescents and young adults. This trend is especially concerning due to the lack of regulations governing the e‐cigarette industry and lack of scientific data on the effects of e‐cigarettes on human health. Using a cultured oral keratinocyte model, we hypothesized that exposure to e‐cigarette vapor would adversely affect the growth of cells that reside in the oral cavity. To test this hypothesis, we cultured the cells with condensate prepared from the vapors of grape flavored e‐cigarette fluids, both with and without nicotine. After three days of treatment, cell numbers were determined using a colorimetric assay. Results indicate that the e‐cigarette condensate adversely affects cell growth, regardless of the absence or presence of nicotine. To understand the underlying mechanisms, changes in gene expression in the cells treated with condensate were evaluated using real‐time qPCR after 24, 48, and 72 hours of treatment. A time‐dependent increase in the levels of CYP1A1 and IL8, gene targets of the aryl hydrocarbon receptor, were observed. Results obtained from similar experiments using the isolated flavoring agent, methyl anthranilate, indicate that formation of a heat‐induced byproduct of this agent may be involved in the observed changes in cell growth. Future experiments will focus on determining the mechanisms by e‐cigarette condensates impact cell growth and identifying the specific chemical byproduct mediating these effects.Support or Funding InformationFunded in part by a grant from the American Society of Pharmacology and Experimental Therapeutics for Summer Undergraduate Research.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Introduction Ceramides are known to show anti-cancer activity. A novel ceramide analog, (S,E)-3-hydroxy-2-(2-hydroxybenzylidene)amino-N-tetradecylpropanamide (analog 315) was developed as part of a larger study focused on finding more effective breast cancer treatments. Objective To assess whether analog 315 shows any or a combination of the following effects in breast cancer cells in vitro: inhibiting proliferation, inducing apoptosis, and altering protein expression. Also, to determine whether it inhibits chemo-resistant breast cancer tumor growth in vivo mouse model. Methods In vitro cell proliferation and apoptosis after treatment with analog 315 were assessed in three breast cancer cell lines (MCF-7, MCF-7TN-R, and MDA-MB-231) and reported. Protein expression was assessed by microarray assay. For the in vivo studies, chemo-resistant breast cancer cells were used for tumor development in two groups of mice (treated and control). Analog 315 (25 mg/kg/day) or control (dimethyl sulfoxide) was administered intraperitoneally for 7 days. Effects of analog 315 on inhibiting the growth of chemo-resistant breast cancer tumors after treatment are reported. Results Analog 315 reduced MCF-7TN-R chemo-resistant tumor burden (volume and weight) in mice. Liver metastasis was observed in control mice, but not in the treated animals. Ki-67, a proliferation marker for breast cancer cells, increased significantly (P < 0.05) in control tumor tissue. In vitro studies showed that analog 315 inhibited cell proliferation, altered protein expression and induced apoptosis in all three breast cancer cell lines studied, of which the effects on MCF-7TN-R cells were the most significant. Conclusion Analog 315 reduced tumor growth in chemo-resistant breast cancer, inhibited cell proliferation, altered protein expression, and induced apoptosis in all three cell lines studied.