Figures and data
Correlation between clinicopathological features and the expression of CPT1A in tumor paraffin section
Aberrant CPT1A mRNA level in CRC.
A. The expression of CPT1A in six GEO microarrays. B. Real-time PCR for CPT1A in 24-paired CRC and adjacent non-tumour tissues. C. Western blot for CPT1A in sixteen-paired CRC and adjacent non-tumour tissues. D. Lower CPT1A mRNA level in COAD than the normal counterparts from TCGA in ualcan database. E. Lower CPT1A mRNA level in READ than the normal counterparts from TCGA in ualcan database. F. IHC assay for CPT1A in two patients. ***, P < 0.001, ** P < 0.01, * P < 0.05.
Correlation of CPT1A with overall survival and neoadjuvant therapy response in rectal cancer patients.
A. IHC assay for CPT1A in two groups of patients, upper with low CPT1A expression and lower with high CPT1A expression. B. The Overall survival (OS) was estimated by the Kaplan-Meier method in rectal cancer patients. C. The OS was estimated by the Kaplan-Meier method in rectal cancer patients in TCGA database. D. IHC assay for CPT1A in two groups of patients, upper with TRG-1 and lower with TRG-2 (TRG means tumour regression grade, AJCC standard, 0, complete response: No remaining viable cancer cells; 1, moderate response: Only small clusters or single cancer cells remaining; 2, minimal response: Residual cancer remaining, but with predominant fibrosis). E. Dot plot showing the IHC score and TRG score of patients. F. Correlation of CPT1A with TRG score, size of dot represents the number. ***, P < 0.001, ** P < 0.01, * P < 0.05.
Radio-sensitivity of stable knock-out or overexpression of CPT1A.
A. The protein level of CPT1A in different groups of cell lines. B. The mRNA level of CPT1A in different groups of cell lines. C. Colony-forming assay of HCT 116-NC and HCT 116-KO cell lines. D. The map of multi-target, single-hit model. E. Colony-forming assay of SW480-RFP and SW480-OE cell lines. F. The map of multi-target, single-hit model. G. Comet assay of different cells. H. Protein expression of γ-H2A.X in HCT 116-NC and HCT 116-KO cell lines. I. Protein expression of γ-H2A.X in SW480-RFP and SW480-OE cell lines. ***, P < 0.001, ** P < 0.01, * P < 0.05.
CPT1A inhibited proliferation and radio-resistance in nude mice.
A. Image of tumours formed in nude mice, with knock out of CPT1A and radiation. B. Scattergram showing the weight of tumours. C. Immunohistochemical staining of Ki67 in tumours. D. Bar chart demonstrating the percentage of positively stained of Ki67 cells. E. Image of tumours formed in nude mice, with overexpression of CPT1A and radiation. F. Scattergram showing the weight of tumours. G. Immunohistochemical staining of Ki67 in tumours. H. Bar chart demonstrating the percentage of positively stained of Ki67 cells. ***, P < 0.001, ** P < 0.01, * P < 0.05.
The effect of CPT1A on ROS related enzyme activity.
A. Enriched KEGG pathway of DEGs in mRNA sequencing. B. Generation and scavenging of ROS in cell. C. ROS of HCT 116-KO cell, SW480-OE cell and their control with DCFH-DA by flow cytometry. D. Bar graph to show the mean of DCFH-DA in HCT 116-KO, HCT 116-NC cells, SW480-NC and SW480-OE cells. E. GSH / GSSG ratio measurement under CPT1A knockout and radiation. F. GSH / GSSG ratio measurement under CPT1A overexpression and radiation. G. Effect of the CPT1A knockout on SOD activity. H. Effect of the CPT1A overexpression on SOD activity. I. Effect of the CPT1A on CAT activity. ***, P < 0.001, ** P < 0.01, * P < 0.05.
CPT1A increases the transcription and protein of ROS scavenge related genes by regulating the transcription factor activity of FOXM1.
A.The protein level of FOXM1, CPT1A, CAT, SOD1, SOD2, SOD3 after knockout and overexpression of CPT1A. B. The mRNA level of FOXM1, CAT, SOD1, SOD2, SOD3 after knockout and overexpression of CPT1A. C. Veen map showing the potential transcription factor of SOD1, SOD2 and CAT. D. The protein level of FOXM1, CPT1A, CAT, SOD1, SOD2 after overexpression of FOXM1 in HCT116-CPT1AKO cells. E. Schematic diagram summarising our working model, namely, decreased CPT1A promotes the transcription factor activity of FOXM1, increasing the mRNA and protein level of CAT, SOD1, and SOD2, followed by increasing ROS scavenge after irradiation and therefore CRC cells become radioresistance. ***, P < 0.001, ** P < 0.01, * P < 0.05.
