Fig. 5: Depletion of MTHFD2 decreases breast cancer cell proliferation and chemotherapy drug resistance. | Communications Biology

Fig. 5: Depletion of MTHFD2 decreases breast cancer cell proliferation and chemotherapy drug resistance.

From: SIRT5-mediated desuccinylation of MTHFD2 enhances chemoresistance in breast cancer cells by reducing therapy-induced senescence

Fig. 5

A Box plots of MTHFD2 transcript levels in BRCA, the RNA-sequencing expression profiles and corresponding clinical information for multi-cancer were downloaded from the TCGA dataset. The statistical significance computed by the Wilcoxon test is annotated by the number of stars (*p-value < 0.05; **p-value < 0.01; ***p-value < 0.001). B Kaplan–Meier survival analysis of the gene signature from TCGA dataset with BRCA based on MTHFD2 expression. C MDA-MB-231 cells were transfected with control or MTHFD2 siRNAs, cell proliferation was analyzed by MTT experiment. D MCF-7 cells were transfected with 3xFlag-Vector or 3xFlag-MTHFD2, cell proliferation was analyzed by EdU-488 experiment. E MDA-MB-231 cells treated with 0, 20 and 40 μM Olaparib for 48 h, and protein expression were analyzed. F MDA-MB-231 cells treated with 0, 0.5 and 1 mM Doxorubicin for 48 h, and protein expression were analyzed. G MDA-MB-231 cells transfected with control siRNA (−) or MTHFD2 siRNA for 48 h, and treated with 50 μM etoposide for 4 h. Protein expression was analyzed by Western blot using the indicated antibodies. Data are representative of three independent experiments. H MCF-7 cells were transfected with control or MTHFD2 siRNAs, γH2AX protein was detected by immunofluorescence. I Effect of ectopic expression of WT MTHFD2, MTHFD2 K44E and Gln132Lys/Asp155Ala-MTHFD2 mutant on cell proliferation. MDA-MB-231 cells were transfected with siMTHFD2 or siControl to knock down the endogenous MTHFD2, and then express FLAG-MTHFD2, FLAG-MTHFD2 K44E, and Gln132Lys/Asp155Ala-MTHFD2. The cell proliferation was assayed by crystal violet staining. J Effect of ectopic expression of WT MTHFD2, MTHFD2 K44E, and Gln132Lys/Asp155Ala-MTHFD2 mutant on DNA damage. MDA-MB-231 cells were transfected with siMTHFD2 or siControl to knock down the endogenous MTHFD2, and then express FLAG-MTHFD2, FLAG-MTHFD2 K44E and Gln132Lys/Asp155Ala-MTHFD2. The γH2AX protein was detected by immunofluorescence. K MDA-MB-231 cells were transfected with control or MTHFD2 siRNAs, and treated with 40 μM Olaparib for 24 h, and cell survival was assayed by crystal violet staining. Protein expression was analyzed by western blot. L MDA-MB-231 cells were transfected with Vector or 3xFlag-MTHFD2, and treated with 40 μM Olaparib, and cell survival was assayed by crystal violet staining. M Effect of ectopic expression of WT MTHFD2, MTHFD2 K44E, and Gln132Lys/Asp155Ala-MTHFD2 mutant on cell chemoresistance. MCF-7 cells were transfected with siMTHFD2 or siControl to knock down the endogenous MTHFD2 and express FLAG-MTHFD2, FLAG-MTHFD2 K44,E and Gln132Lys/Asp155Ala-MTHFD2, and then treated with 40 μM Olaparib for 48 h. The cell proliferation was assayed by EdU-488.

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