Extended Data Fig. 6: CPS1, OTC and ARG1 promote tumour growth, are highly expressed in many human cancers, and inversely correlate with patients survival and p53 expression.
From: p53 regulation of ammonia metabolism through urea cycle controls polyamine biosynthesis
a, Related to Fig. 2g. Western blot analyses of CPS1, OTC, ARG1 and p53 in xenograft tumours derived from p53+/+ and p53−/− HCT116 cells triply transfected with CPS1, OTC and ARG1 siRNAs, or control siRNA are shown. b, Colony formation assay of p53+/+ and p53−/− HCT116 cells triply transfected with CPS1, OTC and ARG1 siRNAs, or control siRNA. Numbers of colonies with a diameter greater than 20 μm were quantified (left). Data are mean ± s.d., n = 6 cultures. **P < 0.01, ***P < 0.001, one-way ANOVA followed by followed by Dunnett’s multiple comparisons test. Representative images of colonies stained with crystal violet at day 14 are shown (right). c, Average weights of xenograft tumours (2 weeks, mean ± s.d., n = 5 tumours in each treatment) generated by p53+/+ HCT116 cells stably expressing CPS1, OTC and ARG1 individually. *P < 0.05, one-tailed t-test. d–f, Box plot comparing transcript levels of CPS1 (d), OTC (e) and ARG1 (f) in liver cancer, leukaemia, prostate cancer, oesophageal adenocarcinoma and their normal counterparts. Data were generated from Oncomine data. In d, liver (maximum = 1.219, median = −0.018, minimum = −2.018, percentile: 90th = 0.709, 75th = 0.348, 25th = −0.423, 10th = −0.88); hepatocarcinoma (maximum = 2.745, median = 0.933, minimum = −0.678, percentile: 90th = 2.269, 75th = 1.638, 25th = 0.449, 10th = −0.005); bone marrow (maximum = −0.997, median = −1.255, minimum = −1.53, percentile: 90th = −0.997, 75th = −0.997, 25th = −1.42, 10th = −1.53); leukaemia (maximum = 2.932, median = 0.298, minimum = −1.391, percentile: 90th = 1.319, 75th = 0.797, 25th = −0.114, 10th = −0.49); oesophagus (maximum = 3.235, median = 1.57, minimum = 0.321, percentile: 90th = 2.446, 75th = 1.878, 25th = 1.266, 10th = 1.076); Barrett’s oesophagus (maximum = 7.71, median = 5.52, minimum = 3.153, percentile: 90th = 7.026, 75th = 6.537, 25th = 4.385, 10th = 3.32). In e, oesophagus (maximum = −0.188, median = -0.314, minimum = −0.581, percentile: 90th = −0.212, 75th = −0.253, 25th = −0.394, 10th = −0.533); Barrett’s oesophagus (maximum = 5.926, median = 4.337, minimum = −0.235, percentile: 90th = 5.42, 75th = 5.217, 25th = 1.976, 10th = 1.542); liver (maximum = 4.132, median = 1.67, minimum = −0.928, percentile: 90th = 3.603, 75th = 3.308, 25th = 0.366, 10th = 0.135); hepatocellular carcinoma (maximum = 4.919, median = 3.918, minimum = −1.121, percentile: 90th = 4.628, 75th = 4.427, 25th = 3.537, 10th = 2.469); CD4-positive T lymphocyte (maximum = −3.268, median = −3.726, minimum = −4.225, percentile: 90th = −3.268, 75th = −3.268, 25th = −3.868, 10th = −4.225); acute adult T cell leukaemia/lymphoma (maximum = −0.441, median = −3.143, minimum = −4.466, percentile: 90th = −1.807, 75th = −2.239, 25th = −3.621, 10th = −3.935). In f, B lymphocyte T lymphocyte (maximum = −2.17, median = −3.004, minimum = −4.31, percentile: 90th = −2.17, 75th = −2.17, 25th = −3.981, 10th = −4.31); centroblast (maximum = −1.586, median = −4.123, minimum = −4.826, percentile: 90th = −1.586, 75th = −1.586, 25th = −4.219, 10th = −4.826); memory B lymphocyte (maximum = −2.355, median = −4.453, minimum = −6.27, percentile: 90th = −2.355, 75th = −2.355, 25th = −5.531, 10th = −6.27); naive pregerminal centre B lymphocyte (maximum = −3.153, median = −4.42, minimum = −5.084, percentile: 90th = −3.153, 75th = −3.153, 25th = −4.439, 10th = −5.084); small cleaved follicle centre cell (maximum = −3.161, median = −3.55, minimum = −5.71, percentile: 90th = −3.161, 75th = −3.161, 25th = −4.39, 10th = −5.71); hairy cell leukaemia (maximum = −3.268, median = −0.612, minimum = −5.659, percentile: 90th = 1.593, 75th = 0.94, 25th = −1.585, 10th = −2.735); prostate (maximum = 0.684, median = −0.708, minimum = −1.331, percentile: 90th = 0.684, 75th = 0.684, 25th = −0.855, 10th = −1.331); benign prostatic hyperplasia (maximum = 1.251, median = 0.92, minimum = 0.25, percentile: 90th = 1.251, 75th = 1.251, 25th = 0.465, 10th = 0.25). n indicates number of samples. g, Kaplan–Meier survival curves of patients with colon cancer based on CPS1 (left), OTC (middle) or ARG1 (right) expression. Data were generated from Gene Expression Omnibus (GEO) ‘the human colon cancer Marisa dataset ’(series GSE39582) via ‘R2: Genomics Analysis and Visualization Platform’. h, Correlation plots of wild-type p53 expression with CPS1, OTC or ARG1 expression with significant Pearson’s correlation in the colon cancer dataset (n = 371 patients) are shown. r, Pearson’s correlation coefficient; x and y axes denote the respective genes being analysed. Data were generated from The Cancer Genome Atlas (TCGA) ‘Liver Hepatocellular Carcinoma’ dataset (ID: LIHC) via ‘R2: Genomics Analysis and Visualization Platform’. See Supplementary Fig. 1 for gel source data.
