Supplementary Figures 1 - 15 from MicroRNA100 Inhibits Self-Renewal of Breast Cancer Stem–like Cells and Breast Tumor Development

Deng, Lu; Shang, Li; Bai, Shoumin; Chen, Ji; He, Xueyan; Martin-Trevino, Rachel; Chen, Shanshan; Li, Xiao-yan; Meng, Xiaojie; Yu, Bin; Wang, Xiaolin; Liu, Yajing; McDermott, Sean P.; Ariazi, Alexa E.; Ginestier, Christophe; Ibarra, Ingrid; Ke, Jia; Luther, Tahra; Clouthier, Shawn G.; Xu, Liang; Shan, Ge; Song, Erwei; Yao, Herui; Hannon, Gregory J.; Weiss, Stephen J.; Wicha, Max S.; Liu, Suling

doi:10.1158/0008-5472.22404500.v1

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Supplementary Figures 1 - 15 from MicroRNA100 Inhibits Self-Renewal of Breast Cancer Stem–like Cells and Breast Tumor Development

journal contribution

posted on 2023-03-30, 22:52 authored by Lu Deng, Li Shang, Shoumin Bai, Ji Chen, Xueyan He, Rachel Martin-Trevino, Shanshan Chen, Xiao-yan Li, Xiaojie Meng, Bin Yu, Xiaolin Wang, Yajing Liu, Sean P. McDermott, Alexa E. Ariazi, Christophe Ginestier, Ingrid Ibarra, Jia Ke, Tahra Luther, Shawn G. Clouthier, Liang Xu, Ge Shan, Erwei Song, Herui Yao, Gregory J. Hannon, Stephen J. Weiss, Max S. Wicha, Suling Liu

Figure S1. Comparison of ALDH-positive population in different breast cancer cell lines and primary xenografts (UM2, MC1, UM1). Figure S2. mir-100 overexpression decreases ki67+ population in vitro. Figure S3. mir-100 overexpression inhibits the proliferation of both ALDH+ population and ALDH- population. Figure S4. mir-100 inhibits tumor growth of SUM149, MCF7 and T47D in NOD/SCID mice. Figure S5. mir-100 expression is higher in the tumor cells from mir-100 group compared to that from CTRL group. Figure S6. The percentage of ALDH+ cells in both primary CTRL and secondary CTRL tumors were increased compared to the parental cells injected. Figure S7. mir-100 expression is higher in the tumor cells from mir-100 induction groups with DOX (Late and Early) compared to that from CTRL group. Figure S8. mir-100 overexpression is necessary to maintain the inhibition of SUM159 tumor growth in NOD/SCID mice. Figure S9. mir-100 knockdown increases tumor growth in vivo. Figure S10. mir-100 inhibits metastasis of Both Total (Unseparated) and ALDH+ SUM159 cells in vivo. Figure S11. Identification of direct targets of mir-100. Figure S12. Knockdown of mir-100 targets mimic the effects of mir-100 overexpression. Figure S13. Verification of SMARCA5 and SMARCD1 level by Western blot. Figure S14. SMARCA5 and SMARCD1 are direct and functional targets of mir-100 in SUM149 cells. Figure S15. Kaplan-Meier survival curves based on the expression of SMARCA5 and SMARCD1.

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ARTICLE ABSTRACT

miRNAs are essential for self-renewal and differentiation of normal and malignant stem cells by regulating the expression of key stem cell regulatory genes. Here, we report evidence implicating the miR100 in self-renewal of cancer stem–like cells (CSC). We found that miR100 expression levels relate to the cellular differentiation state, with lowest expression in cells displaying stem cell markers. Utilizing a tetracycline-inducible lentivirus to elevate expression of miR100 in human cells, we found that increasing miR100 levels decreased the production of breast CSCs. This effect was correlated with an inhibition of cancer cell proliferation in vitro and in mouse tumor xenografts due to attenuated expression of the CSC regulatory genes SMARCA5, SMARCD1, and BMPR2. Furthermore, miR100 induction in breast CSCs immediately upon their orthotopic implantation or intracardiac injection completely blocked tumor growth and metastasis formation. Clinically, we observed a significant association between miR100 expression in breast cancer specimens and patient survival. Our results suggest that miR100 is required to direct CSC self-renewal and differentiation. Cancer Res; 74(22); 6648–60. ©2014 AACR.