Number S7B. S5D. Signaling pathway based on KEGG enrichment analysis of p53-R273H-controlled coding genes in CSC state. Number S5E. GO biological processes enrichment analysis of p53-R273H-controlled coding genes in CSC state. Number S5F. Regulatory network building of TFs (dark blue), lncRNAs (reddish) and mRNAs (light green). The average degree of lncRNAs was 46.39, higher than 35.39, the average degree of protein coding genes. Number S6A. CPI-203 ChIP-qPCR for validating of the binding of p53 and the promotor of lnc273C31 or lnc273C34. Number S6B. The manifestation levels in ALDH positive and ALDH bad cells sorted by FACS. Number S6C. Subcellular localization of lnc273C31 and lnc273C34 was analyzed by RT-qPCR upon biochemical fractionation of p53-R273H speroid cells. Number S7A. Quantitative real-time PCR analyzed the manifestation of stemness-related genes in HCT116 p53 PM cells. Number S7B. Western blot analysis of ZEB1 and snail in lnc273C31 or lnc273C34 depletion cells. Number S8. The association of age (S8A), gender (S8B), smoking (S8C), alcohol abuse (S8D), family history (S8E), lymphatic vessel (S8F), TNM stage (S8G-I) and the manifestation levels of lncRNAs in 25 colorectal malignancy individuals with or without p53-R273H mutation. Table S1. Primers for qPT-PCR. Table S2. Purchased ASO pool sequences. Table S3. Primers for ChIP-qPCR. (DOCX 2058 kb) 13046_2019_1375_MOESM1_ESM.docx (2.0M) GUID:?ADBFA4A9-8C7E-4CA7-BC08-4278DCB0DB85 Additional file 2: The list of differentially expressed lncRNAs. (XLSX 11 kb) 13046_2019_1375_MOESM2_ESM.xlsx (12K) GUID:?CFD1CE10-B3BB-4D14-B2E1-3307BDB92142 Additional file 3: The results of KEGG and GO analysis. (XLSX 18 kb) 13046_2019_1375_MOESM3_ESM.xlsx (18K) GUID:?D48CF9B3-8008-43FC-90DC-CF1CDC226645 Additional file 4: The list of differentially expressed protein coding genes (PCGs). (XLSX 128 kb) 13046_2019_1375_MOESM4_ESM.xlsx (129K) GUID:?BE3F8EAB-BA3E-4943-B8A8-93D556C6FDCD Additional file 5: LncRNA annotation. (XLSX 107 kb) 13046_2019_1375_MOESM5_ESM.xlsx (107K) GUID:?0FE6D367-19A7-434B-A503-761716D7C747 Additional file 6: The list CPI-203 of lncRNAs analyzed by RNA-seq combined with ChIP-seq. (XLSX 29 kb) 13046_2019_1375_MOESM6_ESM.xlsx (30K) GUID:?AF1C5867-293C-450B-B8D7-6053E6BF7D65 Additional file 7: Clinical patient information. (XLSX 11 kb) 13046_2019_1375_MOESM7_ESM.xlsx (11K) GUID:?18429673-4BDC-4CA4-9362-C4CE36FDBBCF Data Availability StatementThe datasets used and/or analyzed during the current study are available from your corresponding author about reasonable request. Abstract Background TP53 is one of the most frequently mutated genes among all malignancy types, and TP53 mutants happen more than 60% in colorectal malignancy (CRC). Among all mutants, you will find three hot places, including p53-R175H, p53-R248W and p53-R273H. Emerging evidence characteristics tumor carcinogenesis to malignancy stem cells (CSCs). Long noncoding RNAs STAT6 (lncRNAs) play important roles in keeping the stemness CPI-203 of CSCs. However, it is unfamiliar if mutant p53-controlled lncRNAs are implicated in the maintenance of CSC stemness. Methods RNA-sequencing (RNA-seq) and ChIP-sequencing CPI-203 (ChIP-seq) were used to trace the lncRNA network controlled by p53-R273H in HCT116 endogenous p53 point mutant spheroid cells generated from the somatic cell knock-in method. RT-qPCR was used to detect lncRNA manifestation patterns, verifying the bioinformatics analysis. Transwell, spheroid formation, fluorescence triggered cell sorter (FACS), xenograft nude mouse model, tumor rate of recurrence assessed by intense limiting dilution analysis (ELDA), Western blot assays and chemoresistance analysis were performed to elucidate the functions and possible mechanism of lnc273C31 and lnc273C34 in malignancy stem cells. Results p53-R273H exhibited more characteristics of CSC than p53-R175H and p53-R248W. RNA-seq profiling recognized 37 up? controlled and 4 down controlled differentially indicated lncRNAs controlled by p53-R273H. Combined with ChIP-seq profiling, we further verified two lncRNAs, named CPI-203 as lnc273C31 and lnc273C34, were essential in the maintenance of CSC stemness. Further investigation illustrated that lnc273C31 or lnc273C34 depletion dramatically diminished colorectal malignancy migration, invasion, malignancy stem cell self-renewal and chemoresistance in vitro. Moreover, the absence of lnc273C31 or lnc273C34 dramatically delayed tumor initiation and tumorigenic cell rate of recurrence in vivo. Also, lnc273C31 and lnc273C34 have an impact on epithelial-to mesenchymal transition (EMT). Finally, lnc273C31 and lnc273C34 were significantly highly indicated in CRC cells with p53-R273H mutation compared to those with wildtype p53. Conclusions The present study unveiled a high-confidence arranged.