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. 2021 Oct 28;49(20):11708–11727. doi: 10.1093/nar/gkab976

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© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 8. — RBM6 deficiency is correlated with breast cancer progression and confers sensitivity to cisplatin in mice xenograft. (A, B) Cisplatin treatment hypersensitizes MCF10A RBM6-KO1 (A) and MDA231 RBM6-shRNA#1 (B) cells. Cell viability was assessed by the CellTiter 96^® proliferation assay. Data presented are mean of three independent experiments ± SD. (C, D) MDA231 xenograft tumor model. 2.5 × 10⁶ MDA231 cells expressing either scramble or shRNA against RBM6 (RBM6 shRNA#1) were injected subcutaneously. When tumors reached ∼100 mm³, mice were assigned randomly to either control (vehicle: PBSx1) or cisplatin (5 mg/kg in PBSx1) treatment (n = 8 for each group). Mice were euthanized when the tumors of control mice reached ∼1500 mm³. Left: Tumors were collected and photographed. Scale bar, 1 cm. Right: Average tumor weight ± SD are shown. (E, F) γH2AX and cleaved caspase 3 (c.caspase 3) in control and RBM6-deficient MDA231 xenografts. Graph shows the percentage of γH2AX (E) and c.caspase 3 (F) positive cells. Data presented are the mean ± SEM of nine different fields from three different tumor sections for each group. (G) RBM6 protein expression from quantitative proteome analysis of 375 cancer cell lines from different types of tumors. Data are presented as min to max box plot of the relative protein expression compared to the mean protein expression across all cancer types. The number of cell lines from each cancer type is indicated below. The statistical significance of the differences in protein expression in the indicated cancer types relative to the mean protein expression was determined using one-sample Wilcoxon test. (H) As in (G) except that cell lines representing primary and metastatic breast cancer were used for the analysis. (I) Representative images (top) and IHC analysis (bottom) of human breast cancer tissue microarray (TMA) (US Biomax – BR1005b, BR1008b and BR2082c) showing immunohistochemically staining of RBM6 in normal, primary and metastatic breast tissue samples. *P< 0.01, **P < 0.001, ***P < 0.0001, ^****P < 0.00001.