Journal: International Journal of Oncology

Article Title: Smoking promotes colorectal cancer via the CKAP2L/AREG axis

doi: 10.3892/ijo.2026.5872

Figure Lengend Snippet: CKAP2L promotes proliferation and migration of CRC cells by promoting AREG expression. (A) RNA sequencing results for transduced CRC cells. (B) Venn diagram shows the overlapping genes between transduced cells and CSE-treated HCT116 cells. (C) Relative expression of AREG in CRC cells treated with CSE detected by RT-qPCR and normalized against β-actin. (D) Relative expression levels of AREG and CKAP2L were detected by RT-qPCR and normalized against β-actin. (E) Secreted protein levels of AREG were detected by enzyme-linked immunosorbent assay. (F) Proliferation of transduced/transfected cells was measured by Cell Counting Kit 8. (G) Migration of transduced/transfected cells was measured by Transwell assay (×100 magnification). (H) Expression levels of proteins were measured by western blotting. Data were analyzed using two-way ANOVA followed by Tukey test, or unpaired Student's t-test for two groups. * P<0.05, ** P<0.01, *** P<0.001 and **** P<0.0001. AREG, amphiregulin; CKAP2L, cytoskeleton-associated protein 2-like; CRC, colorectal cancer; CSE, cigarette smoke extract; EGFR, epidermal growth factor receptor; NC, negative control; OE, overexpression; p-, phosphorylated; RT-qPCR, reverse transcription-quantitative PCR; sh, short hairpin; si, small interfering.

Article Snippet: The concentration of secreted AREG in the culture medium was then measured using the Human AREG ELISA Kit (cat. no. EK0304; Wuhan Boster Biological Technology, Ltd.) according to the manufacturer's instructions.

Techniques: Migration, Expressing, RNA Sequencing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Transfection, Cell Counting, Transwell Assay, Western Blot, Negative Control, Over Expression, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Oncology

Article Title: Smoking promotes colorectal cancer via the CKAP2L/AREG axis

doi: 10.3892/ijo.2026.5872

Figure Lengend Snippet: CKAP2L promotes proliferation and migration of colorectal cancer cells through the STAT3/AREG/EGFR axis. (A) Expression levels of STAT3 and p-STAT3 proteins measured by western blotting. (B) Calculation of IC 50 in HCT116 cells treated with Stattic for 24 h. (C) Levels of AREG were measured by reverse transcription-quantitative PCR and enzyme-linked immunosorbent assay. (D) Migration of cells was detected by Transwell assay (×100 magnification). (E) Proliferation of cells was measured by Cell Counting Kit 8. (F) Expression levels of proteins measured by western blotting. (G) Binding peak of STAT3 on the promoter region of AREG was detected by Cistrome Data Browser, the binding motif was predicted using the JASPAR database, and the binding of STAT3 to the AREG promoter was evaluated by chromatin immunoprecipitation assay. Data were analyzed using (A and G) Unpaired Student's t-test, and (C-F) two-way ANOVA followed by Tukey test. * P<0.05, ** P<0.01, *** P<0.001 and **** P<0.0001. AREG, amphiregulin; CKAP2L, cytoskeleton-associated protein 2-like; EGFR, epidermal growth factor receptor; NC, negative control; p-, phosphorylated; sh, short hairpin; STAT3, signal transducer and activator of transcription 3; TSS, transcription start site.

Article Snippet: The concentration of secreted AREG in the culture medium was then measured using the Human AREG ELISA Kit (cat. no. EK0304; Wuhan Boster Biological Technology, Ltd.) according to the manufacturer's instructions.

Techniques: Migration, Expressing, Western Blot, Reverse Transcription, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Transwell Assay, Cell Counting, Binding Assay, Chromatin Immunoprecipitation, Negative Control