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. 2021 Aug 24;12:5103. doi: 10.1038/s41467-021-25354-4

Fig. 5. GPX4 inhibition sensitizes 27HCR cells to ferroptosis and reduces their metastatic capability.

Fig. 5

a A total of 2 × 105 luciferase labeled Py230-27HCS and -27HCR cells were injected intravenously into nude or C57/BL6 mice. After 2 months, Py230-27HCS cells developed micrometastasis and Py230-27HCR cells formed overt macrometastasis. Bioluminescent images of lung metastasis as shown in the left panels. Metastatic tumor cells were then recovered from lungs and propagated as cell lines (Micrometastasis: Micro. Met1 and 2 and Macrometastasis: Macro Met1 and 2). Py230 parental and lung metastasis derivative cell lines were treated with GPX4 inhibitors, RSL3 (middle), and ML210 (right) for 48 h, followed by cell proliferation assay. Relative cell growth (% of Growth) was calculated by normalizing each treatment condition to control-treated wells. Data plotted as mean ± SEM from five technical replicates. b 2 × 105 GPX4 wild-type (shSCR) and knockdown (KD2 and KD4) B16F10-27HCS and -27HCR cells were injected intravenously into C57BL/6 mice (n = 5 mice). Lung metastases were graded and scored when mice were euthanized on Day 16. Representative images of lung tissue used for grading are shown in the upper panel. c 3 × 105 GPX4 wild-type (shSCR) and knockdown (KD2 and KD4) Py230-27HCR cells, as well as GPX4 wild-type (shSCR) Py230-27HCS cells were injected intravenously into C57BL/6 mice (n = 5 mice). Lung metastases were quantified when mice were euthanized on Day 40. Representative lung images are shown in the upper inset. Data plotted as mean ± SEM; P values were calculated using two-way ANOVA. d The proposed model to explain how cancer cells respond to 27HC treatment. Acute (left) treatment with 27HC disrupts lipid metabolism via interfering with SREBPs and LXR signaling, and this results in the inhibition of cell growth and migration. Cancer cells can adapt to the metabolic stress imposed by chronic treatment by 27HC (right). The cells that survive (27HC resistant cells) increase lipid uptake and accommodate the metabolic stress associated with this activity by upregulating the activity of processes that allow them to withstand lipid oxidative stress (ferroptosis); an activity which confers upon them enhanced tumor growth and metastatic capabilities. Numerical source data are reported in the Source Data File.