Fig. 3: Smyd3 is negatively regulated by Brca1 but positively regulated by ERα in mammary tumor initiation and progression. | Cell Death & Disease

Fig. 3: Smyd3 is negatively regulated by Brca1 but positively regulated by ERα in mammary tumor initiation and progression.

From: Oncogenic activation of SMYD3-SHCBP1 promotes breast cancer development and is coupled with resistance to immune therapy

Fig. 3

a The expression pattern of Smyd3 in WT and Brca1 MT mammary epithelial cell lines at 0 and 24 hours after 100 nM estradiol (E2) treatment as determined by qPCR. b The expression of Smyd3, and Psmd9 under E2 treatment for 2 hours in WT-B477 (WT) and Brca1-MT G600 (MT) cells. c The Expression of Esr1, Pgr, and Ncoa1 after 24 hours of 100 nM E2 treatment as determined by qPCR in B477 (WT Brca1) and G600 (Brca1-MT) cell lines. d Luciferase promotor activity assay from different fragments of Smyd3 promoters, including the fragment from -1500 bps to + 760 bps (WTP2), -1160 bps to +760 bps serve as (WTP1), -800 bps to +760 bps (MTP1), -600 bps to + 760 bps (MTP2), -400 bps to +760 bps (MTP3), -250 bps to +760 bps (MTP4), -100 bps to +760 bps (MTP5), -50 bps to +760 bps (MTP6), and -1160 bps to -288 bps (MTP7). e Luciferase activity assay using WTP1 and MTP7 of the Smyd3 promoter under E2 treatment or overexpression of mBrca1 cDNA in both WT B477 and Brca1 MT G600 mammary epithelial cell lines. f Kaplan-Meier tumor-free curve of Brca1Δ11/Δ11;p53+/− under virgin (n = 22) or parous (n = 23) conditions. Virgin Brca1Δ11/+;p53+/− mice (n = 15 mice) were also set up as controls and only one of them developed lymphoma at about 7 months of age. All animals were kept under the same house condition. The parous mice had gone through two full-term of pregnancies. The survival free mice starting from 35 weeks between Brca1 mutant virgin and parous mice is significantly different by Fisher’s exact test (p = 0.017). g Comparison of mammary tumor incidence after experiencing two times pregnancies (14/23, 60%) vs virgin (3/22, 13.5%) in Brca1Δ11/Δ11;p53+/− mice by Fisher’s exact test (p = 0.002). h The tumor growth curve of nude mice with implantation of WT B477 cells in both virgin and pregnant groups (n = 8 mice/group). i The tumor growth curve of nude mice with implantation of MT G600 cells in both virgin and pregnant groups (n = 7 mice/group). j The plot of tumor weight at Day 22 after fat pad implantation in nude mice of B477 and G600 cells in both virgin groups and groups with experience of pregnancy from the same cohort of mice in h and i. k The protein levels of Smyd3, ERα, and H3K4me3 in tumor tissues of the same cohort of mice in j. l Summary of a signaling pathway from Brca1 deficient mice.

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