Fig. 7: SB-T-101141 promotes lipid peroxidation and inhibits CISD1 expression via KHSRP. | Cell Death & Disease

Fig. 7: SB-T-101141 promotes lipid peroxidation and inhibits CISD1 expression via KHSRP.

From: A novel taxane SB-T-101141 triggers a noncanonical ferroptosis to overcome Paclitaxel resistance of breast cancer via iron homeostasis-related KHSRP

Fig. 7

A–D Iron (A) and ferrous ions (B) detections, Prussian blue staining (C), and MDA detection (D) in the KHSRP knock-down and parental MCF-7 cells. Cells were pretreated with DFOM (50 μΜ) for 1 h, followed by SB-T-101141 (3 μM) treatment for 24 h. Results were analyzed using the student t test (mean ± SD, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001; bar indicates 100 μm. E Lipid ROS detection in MCF-7 cells with the BODIPY 581/591 C11 probes (top panel). Cells were transiently transfected with KHSRP-specific siRNAs (si-KHSRP) for 72 h along with the scramble control (si-NC) and then treated with SB-T-101141 (3 μM) for 16 h. Results were analyzed (bottom panel) using the student t test (mean ± SD, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. F Analysis of ferroptosis-related genes from the genome-scale CRISPR-Cas9 screening of SB-T-101141 targets in breast cancer cells. The red rectangle indicates the top two genes associated with ferroptosis. G, H Kaplan–Meier plots of CISD1 (G) and CAT (H) in the survival of breast cancer patients using the website tool (http://kmplot.com/). The high expression of CISD1 or CAT is in red, and the low expression is in black. I, J qRT-PCR detection of CAT or CISD1 in MCF-7 (I) and MCF-7PR (J) cells. Cells were pretreated with DFOM for 1 h, and then with SB-T-101141 (MCF-7: 3 μM; MCF-7PR: 1.5 μM) for 16 h. Results were analyzed using the student t test (mean ± SD, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. K, L Immunoblots of CAT or CISD1 in MCF-7 (K, top panel) and MCF-7PR (L, top panel) cells treated as (I, J) at different time points. The results were quantified (bottom panel) and analyzed using the student t test (mean ± SD, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. M qRT-PCR detection of CISD1 in KHSRP knock-down (si-KHSRP) MCF-7 cells. Cells were transfected with the specific siRNA for 72 h, pretreated with DFOM (100 μM) for 1 h, and then with SB-T-101141 (3 μM) for 16 h. Results were analyzed using the student t test (mean ± SD, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001. N Immunoblots (left panel) of the indicated proteins in KHSRP knock-down (si-KHSRP) MCF-7 cells treated as (M). The results were quantified (right panel) and analyzed using the student t test (mean ± SD, n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

Back to article page