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. 2021 Apr 1;35(7-8):528–541. doi: 10.1101/gad.340505.120

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© 2021 Tang et al.; Published by Cold Spring Harbor Laboratory Press

This article, published in Genes & Development, is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 4. — Depletion of BIRC5 attenuates Trp53^R172H-driven lung metastasis. (A,B) SURVIVIN expression is depleted or decreased significantly in Trp53^R172H/− or Trp53^−/− cells with two independent shRNAs (shBIRC5.1 and shBIRC5.2). Relative intensity densitometry of SURVIVIN results are shown at bottom. (C,D) BIRC5 depletion in Trp53^R172H/− or Trp53^−/− cells is confirmed by qPCR. Trp53^R172H/− BIRC5 Ctrl versus shBIRC5.1 versus shBIRC5.2, n = 3 per group, (**) P = 0.0021 Trp53^R172H/− BIRC5 Ctrl versus shBIRC5.1, (**) P = 0.0015 Trp53^R172H/− BIRC5 Ctrl versus shBIRC5.2, one-way ANOVA. Trp53^−/− BIRC5 Ctrl versus shBIRC5.1 versus shBIRC5.2, n = 3 per group. Error bars represent SEM. (E,F) Depletion of BIRC5 significantly decreased cell invasion in Trp53^R172H/− but not Trp53^−/− cells. Trp53^R172H/− BIRC5 Ctrl versus shBIRC5.1 versus shBIRC5.2, n = 9 per group, (****) P < 0.0001 Trp53^R172H/− BIRC5 Ctrl versus shBIRC5.1, (****) P < 0.0001 Trp53^R172H/− BIRC5 Ctrl versus shBIRC5.2, one-way ANOVA. Error bars represent SEM. (G,H) BIRC5 depletion significantly reduces subcutaneous tumor growth at early stage. Trp53^R172H/− BIRC5 Ctrl (n = 6 flanks) versus shBIRC5.1 (n = 8 flanks) versus shBIRC5.2 (n = 6 flanks). (**) P = 0.0042 day 8; (*) P = 0.00214 day 11; (***) P = 0.0004 day 13; (*) P = 0.0261 day 17, one-way ANOVA. Error bars represent SEM. Trp53^−/− BIRC5 Ctrl (n = 6 flanks) versus shBIRC5.1 (n = 6 flanks) versus shBIRC5.2 (n = 6 flanks). (*) P = 0.0322 day 10; (**) P = 0.0016 day 12, one-way ANOVA. Error bars represent SEM. (I) Depletion of BIRC5 in Trp53^R172H/− ESCC cells significantly reduces lung metastasis burden number detected at week 8 after tail-vein injection. BIRC5 Ctrl (n = 6) versus shBIRC5.1 (n = 5), P = 0.0057; BIRC5 Ctrl (n = 6) versus shBIRC5.2 (n = 4), P = 0.0009, one-way ANOVA. Error bars represent SEM. Scale bar, 1 mm. (J) Depletion of BIRC5 in Trp53^−/− ESCC cells does not significantly reduce lung metastasis burden number detected at week 8 after tail-vein injection. BIRC5 Ctrl (n = 4) versus shBIRC5.1 (n = 3) versus shBIRC5.2 (n = 4). Error bars represent SEM. Scale bar, 1mm.