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. 2019 May 31;9(13):3879–3902. doi: 10.7150/thno.31716

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© Ivyspring International Publisher

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) license (https://creativecommons.org/licenses/by-nc/4.0/). See http://ivyspring.com/terms for full terms and conditions.

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FOXK2 promotes CRC metastasis by upregulating EGFR expression. (A) RT-qPCR and Western blotting analysis of FOXK2 and EGFR expression in CRC cells. n = 3 independent experiments performed in triplicate. * P < 0.05 compared with the control. The data are presented as the mean±s.d. (B) Luciferase reporter assays of the indicated cells cotransfected with pCMV-FOXK2 and the EGFR promoter luciferase construct. n = 3 independent experiments performed in triplicate. * P < 0.05; ** P < 0.01 compared with the control. The data are presented as the mean±s.d. (C) Relative luciferase activity was determined after serially truncated and mutated EGFR promoter constructs were cotransfected with pCMV-FOXK2. ChIP-qPCR assays demonstrated that FOXK2 directly binds to the EGFR promoter in human CRC tissues and CRC cells. * P < 0.05 compared with the control. The data are presented as the mean±s.d. (D) In vivo lung metastatic assays. Cells were injected into the tail veins of mice (n = 10 mice per group). Bioluminescence imaging 9 weeks after implantation, the incidence of lung metastasis, overall survival times, the number of metastatic lung nodules, and H&E staining of lung tissues from the different groups are shown. Scale bars: top, 500 μm; bottom, 40 μm. * P < 0.05 compared with the control. The data are presented as the mean±s.d. (E) In vivo liver metastasis assays. Cells were injected into the spleens of mice (n = 10 mice per group). Bioluminescence imaging 9 weeks after implantation, the incidence of liver metastasis, overall survival times, the number of metastatic liver nodules, and H&E staining of liver tissues from the different groups are shown. Scale bars: top, 200 μm; bottom, 40 μm. * P < 0.05 compared with the control. The data are presented as the mean±s.d.

FOXK2 promotes CRC metastasis by upregulating EGFR expression. (A) RT-qPCR and Western blotting analysis of FOXK2 and EGFR expression in CRC cells. n = 3 independent experiments performed in triplicate. * P < 0.05 compared with the control. The data are presented as the mean±s.d. (B) Luciferase reporter assays of the indicated cells cotransfected with pCMV-FOXK2 and the EGFR promoter luciferase construct. n = 3 independent experiments performed in triplicate. * P < 0.05; ** P < 0.01 compared with the control. The data are presented as the mean±s.d. (C) Relative luciferase activity was determined after serially truncated and mutated EGFR promoter constructs were cotransfected with pCMV-FOXK2. ChIP-qPCR assays demonstrated that FOXK2 directly binds to the EGFR promoter in human CRC tissues and CRC cells. * P < 0.05 compared with the control. The data are presented as the mean±s.d. (D) In vivo lung metastatic assays. Cells were injected into the tail veins of mice (n = 10 mice per group). Bioluminescence imaging 9 weeks after implantation, the incidence of lung metastasis, overall survival times, the number of metastatic lung nodules, and H&E staining of lung tissues from the different groups are shown. Scale bars: top, 500 μm; bottom, 40 μm. * P < 0.05 compared with the control. The data are presented as the mean±s.d. (E) In vivo liver metastasis assays. Cells were injected into the spleens of mice (n = 10 mice per group). Bioluminescence imaging 9 weeks after implantation, the incidence of liver metastasis, overall survival times, the number of metastatic liver nodules, and H&E staining of liver tissues from the different groups are shown. Scale bars: top, 200 μm; bottom, 40 μm. * P < 0.05 compared with the control. The data are presented as the mean±s.d.