Additional file 1 of MiR-181a targets STING to drive PARP inhibitor resistance in BRCA- mutated triple-negative breast cancer and ovarian cancer
Creators: name: Bustos, Matias A.
name: Yokoe, Takamichi
name: Shoji, Yoshiaki
name: Kobayashi, Yuta
name: Mizuno, Shodai
name: Murakami, Tomohiro
name: Zhang, Xiaoqing
name: Sekhar, Sreeja C.
name: Kim, SooMin
name: Ryu, Suyeon
name: Knarr, Matthew
name: Vasilev, Steven A.
name: DiFeo, Analisa
name: Drapkin, Ronny
name: Hoon, Dave S. B.
Publisher: name: figshare
Year: 2023
Subjects: subject: Biochemistry
subject: Cell Biology
subject: Genetics
subject: FOS: Biological sciences
subject: Molecular Biology
subject: Pharmacology
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Immunology
subject: FOS: Clinical medicine
subject: Biological Sciences not elsewhere classified
subject: Cancer
subject: Hematology
subject: Plant Biology

description: Additional file 1: Material and Methods. Figure S1. TNBC olaparib-resistant cell lines and miR-181a overexpressing cell lines showed cross-resistance to cisplatin. A Schematic representation of the seed sequences for miR-181a-5p, miR-181b-5p, miR-181c-5p, and miR-181d-5p in the 3′UTR of TMEM173 gene. B–C. Drug sensitivity assays comparing parental and olaparib-resistant (OlaR) HCC1937 cell line treated with different concentrations of olaparib (B) or cisplatin (C) (Two-way ANOVA and Sidak’s multiple comparisons test). D Cell viability assays comparing parental and OlaR HCC1937 cell line (Two-way ANOVA and Sidak’s multiple comparisons test). E Representative images for GFP positive cells with miR-181a-OV in MDA-MB-436, HCC1395, and HCC1937 cell lines using fluorescence microscopy. Scale bars: 100 µm. F–H Cell viability assays comparing miR-181a-OV and empty vector (CTRL) in MDA-MB-436 (E), HCC1395 (F), and HCC1937 (G) cell lines (two-way ANOVA and Sidak’s multiple comparisons test). I Cell viability assays comparing miR-181a-OV and empty vector (CTRL) HCC1937 cell lines treated with different concentrations of cisplatin (two-way ANOVA and Sidak’s multiple comparisons test). Cell viability assays were performed in triplicates. Figure S2. STING overexpression or downregulation does not affect cell proliferation. A Representative images for STING-OV (GFP) in MDA-MB-436, HCC1395, and HCC1937 cell lines using fluorescence microscopy. Scale bars: 100 µm. B–D Cell viability assays comparing STING-OV and empty vector (CTRL) in MDA-MB-436 (B), HCC1395 (C), and HCC1937 (D) cell lines (two-way ANOVA and Sidak’s multiple comparisons test). E–G Cell viability assays comparing si-STING and si-CTRL in MDA-MB-436 (E), HCC1395 (F), and HCC1937 (G) (two-way ANOVA and Sidak’s multiple comparisons test). Figure S3. STING is downregulated in TNBC and relates to outcomes. A STING mRNA levels in normal breast (Normal) and primary BC (Primary) tissues in the TCGA and GTEx databases (Student’s t-test). B STING mRNA levels in tissues from tumor-adjacent normal breast (Adj. Normal), Luminal (Lum), Luminal-HER2 (Lum-HER2), HER2, and TNBC in the TCGA BRCA dataset (One-way ANOVA and Tukey’s multiple comparisons test). C STING mRNA levels in tissues from the tumor-adjacent normal breast (Adj. Normal), Normal-like (Norm-like), Luminal-A (LumA), Luminal-B (LumB), HER2-enriched, and basal-like breast cancer (BLBC) in the TCGA BRCA dataset (One-way ANOVA and Tukey’s multiple comparisons test). D–F Survival analysis of RFS (D), OS (E), and DMFS (F) for patients with TNBC in the TCGA, GEO, and EGA databases combined (Log-rank test). G–I Survival analysis of RFS (G), OS (H), and DMFS (I) for patients with BLBC in the TCGA, GEO, and EGA database combined (Log-rank test). *p&lt;0.05, **p &lt;0.01, ***p &lt;0.001. Figure S4. Analysis of the mRNA levels of the downstream components of the STING pathways. A–E Quantification by RT-qPCR of STING (A), IL6 (B), IFNB (C), IL12A (D), and IL12B (E) mRNA levels in empty vector (CTRL) and STING-OV HCC1395 cell line (Student’s t-test). F–J Quantification by RT-qPCR of STING (F), IL6 (G), IFNB (H), IL12A (I), and IL12B (J) mRNA levels in empty vector (CTRL) and STING-OV MDA-MB-436 cell line (Student’s t-test). K–O Quantification by RT-qPCR of STING (K), IL6 (L), IFNB (M), IL12A (N), and IL12B (O) mRNA levels in miR-181a-OV and CTRL in HCC1395 cell line (Student’s t-test). P-T Quantification by RT-qPCR of STING (P), IL6 (Q), IFNB (R), IL12A (S), and IL12B (T) mRNA levels in miR-181a-OV and empty vector (CTRL) in MDA-MB-436 cell line (Student’s t-test). *p&lt;0.05, **p &lt;0.01, ***p &lt;0.001. Figure S5. Correlation analysis between miR-181a levels and drug activities. A–D Correlation between miR-181a levels and cisplatin (A), rucaparib (B), olaparib (C), talazoparib (D) drug activity in the BC cell lines obtained from CCLE and GDSC BRCA datasets (Pearson’s correlation coefficient). F–I Correlation between miR-181a levels and cisplatin (F), rucaparib (G), olaparib (H), talazoparib (I) drug activity in the OvCa cell lines obtained from CCLE and GDSC BRCA datasets (Pearson’s correlation coefficient). Figure S6. Uncropped western blotting images. A-C Uncropped western blotting images for Figures 3D (A), H (B), and I (C) are shown. Figure S7. Uncropped western blotting images. A–D. Uncropped western blotting images for Figures 3J (A), K (B), 4A (C), and E (D) are shown. Figure S8. Uncropped western blotting images. A Uncropped western blotting images for Figure 6A (A) and G (B) are shown.