Fig. 4: VSTM2L inhibition sensitizes PCa cells to ferroptosis inducer, RSL3.
A Viability of 22Rv1 or DU145 cells transfected with VSTM2L shRNA or control shRNA was measured by CCK-8 in the presence of varying concentrations of RSL3 (0–100 μM) for 24 h. Data shown as mean ± SD of n = 3 biological replicates. B The schematic diagram of the in vivo experimental process. 1 × 106 22Rv1 shVSTM2L cells and control cells were inoculated trans-subcutaneous into nude mice. RSL3 (5 mg/kg) was intraperitoneally injected into the mice on alternate days for a total of six injections. The tumor tissues were isolated on 27th days post-inoculation. C, D General view of tumor weight (Control+RSL3 VS shVSTM2L + RSL3 P = 6.6076E-7) (C) and tumor volume (D) (examined every 3 days) of each indicated group at 27 days after cell injection. Data shown as mean ± SD, n = 5 tumors, ns, not significant. E, F The levels of GSH (E) and BH4 (Control VS shVSTM2L P = 0.000018) (F) in the tumor tissues at the indicated endpoint. Data shown as mean ± SD, n = 5 tumors. G-K Immunohistochemistry (IHC) and hematoxylin and eosin (H & E) staining for Ki-67 (Control VS shVSTM2L P = 0.000005, Control+RSL3 VS shVSTM2L + RSL3 P = 4.1427E-11) (G, H), VSTM2L (Control VS shVSTM2L P = 0.000014, Control+RSL3 VS shVSTM2L + RSL3 P = 0.000011) (G, I), GPX4 (Control VS shVSTM2L P = 0.000063, Control+RSL3 VS shVSTM2L + RSL3 P = 5.0408E-11) (G, J) and C-Caspase3 (G, K) were performed in isolated tumor tissues. Data shown as mean ± SD, n = 5 tumors, and each tumor slice randomly selected 5 magnification fields, Scale bar =20 μm. P value was determined by two-way ANOVA (A, D) and unpaired two-tailed Student’s t-test without adjustments (C, E, F, H-K). Source data are provided as a Source Data file.
