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. 2021 May 24;13(10):14015–14038. doi: 10.18632/aging.203012

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Copyright: © 2021 He et al.

This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Depletion of NUPR1 promoted sensitivity to sorafenib in ccRCC. (A) Time and concentration-dependent manner of sorafenib treatment on NUPR1 expression in ccRCC cell lines verified by qRT-PCR. (B) Western blot assay of sorafenib inducing NUPR1 protein expression. (C) CCK-8 assay of NUPR1 silencing after sorafenib treatment at the indicated concentrations for 24h. The IC50 values were 10.26, 5.28, 5.86 μM for shCtrl, shNUPR1-1, shNUPR1-2, respectively in Caki-2. The IC50 values were 8.73, 4.55, 4.36 μM, respectively in A498. (D) Colony formation experiments of NUPR1 silencing after sorafenib (5μM) treatment for 2 weeks. (E) Flow cytometry analysis of effects of NUPR1 depletion on apoptosis after sorafenib (5μM) treatment for 24h. (F) Growth curves of subcutaneous xenografts in nude mice (n=5) under different treatments. (G) Images of nude mice and anatomical picture of subcutaneous xenografts. (*p < 0.05, **p < 0.01, ***p < 0.001). CCK-8: cell counting kit-8; ccRCC: clear cell renal cell carcinoma; IC50: 50% inhibiting concentration; qRT-PCR: quantitative real-time reverse transcription PCR.