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Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance

  • Authors:
    • Eunyoung Tak
    • Minhee Kim
    • Youngra Cho
    • Sueun Choi
    • Jihun Kim
    • Buhm Han
    • Hyung-Don Kim
    • Chloe Soo‑Hyun Jang
    • Jeong Eun Kim
    • Yong Sang Hong
    • Sun Young Kim
    • Tae Won Kim
  • View Affiliations / Copyright

    Affiliations: Department of Convergence Medicine, Asan Medical Center, University of Ulsan College of Medicine, Songpa, Seoul 05505, Republic of Korea, Asan Medical Institute of Convergence Science and Technology (AMIST), Asan Medical Center, University of Ulsan College of Medicine, Songpa, Seoul 05505, Republic of Korea, Department of Pathology, Asan Medical Center, University of Ulsan College of Medicine, Songpa, Seoul 05505, Republic of Korea, Department of Biomedical Sciences, Seoul National University College of Medicine, Jongro, Seoul 03080, Republic of Korea, Department of Oncology, Asan Medical Center, University of Ulsan College of Medicine, Songpa, Seoul 05505, Republic of Korea
    Copyright: © Tak et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 15
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    Published online on: November 12, 2021
       https://doi.org/10.3892/or.2021.8226
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Abstract

Neurofibromin 1 (NF1) is a tumor suppressor that has been previously reported to regulate RAS‑MAPK signaling. The present study investigated the possible relationship between NF1 expression and anti‑EGFR antibody (cetuximab) sensitivity in colorectal cancer cell lines. In addition, primary or metastatic colorectal cancer samples from patients treated with cetuximab were assessed for the association of cetuximab sensitivity. The quantities of the NF1 transcript, NF1‑related pathway enrichment and NF1 mutation profile were measured and investigated using RNA sequencing and targeted DNA sequencing. Based on growth inhibition and colony formation assay results, cell lines were designated to be cetuximab‑sensitive (NCI‑H508 and Caco2) or cetuximab‑resistant (KM12C and SM480). Western blotting revealed NF1 was highly expressed in cetuximab‑sensitive cell lines whilst there was little expression in their cetuximab‑resistant counterparts. Knocking down NF1 expression using small interfering RNA in the cetuximab‑sensitive cell lines enhanced the phosphorylation of MEK and ERK according to western blotting. NF1 knockdown also reduced apoptosis, as observed by the decreased number of apoptotic bodies by DAPI nuclear staining and reduced cleavage of caspase and poly‑(ADP ribose) polymerase. NF1 overexpression by transfection with GTPase‑activating protein‑related domain subunit rendered the cetuximab‑resistant cell lines, KM12C and SW480, more susceptible to cetuximab‑induced apoptosis. RNA sequencing of 111 RAS and BRAFV600 wild‑type tumor samples collected from cetuximab‑treated patients with metastatic colorectal cancer revealed that the pre‑treatment NF1 expression levels were not associated with the cetuximab response. However, tumor samples obtained after cetuximab treatment displayed slightly lower NF1 transcript levels compared with those in the pre‑treatment samples, suggesting that exposure to the anti‑EGFR antibody may be associated with reduced NF1 expression levels. Next‑generation sequencing revealed that the frequency of inactivating mutations in NF1 were rare (1.8%) in patients with colorectal cancer and were not associated with the protein expression levels of NF1 except for in a small number of cases (0.5%), where the biallelic inactivation of NF1 was observed. To conclude, the present study showed that modification of NF1 expression can affect sensitivity to cetuximab in colorectal cancer cell lines, though a limitation exists in terms of its potential application as a biomarker for RAS and BRAFV600 wild‑type tumors.
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Copy and paste a formatted citation
Spandidos Publications style
Tak E, Kim M, Cho Y, Choi S, Kim J, Han B, Kim H, Jang CS, Kim JE, Hong YS, Hong YS, et al: Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance. Oncol Rep 47: 15, 2022.
APA
Tak, E., Kim, M., Cho, Y., Choi, S., Kim, J., Han, B. ... Kim, T.W. (2022). Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance. Oncology Reports, 47, 15. https://doi.org/10.3892/or.2021.8226
MLA
Tak, E., Kim, M., Cho, Y., Choi, S., Kim, J., Han, B., Kim, H., Jang, C. S., Kim, J. E., Hong, Y. S., Kim, S. Y., Kim, T. W."Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance". Oncology Reports 47.1 (2022): 15.
Chicago
Tak, E., Kim, M., Cho, Y., Choi, S., Kim, J., Han, B., Kim, H., Jang, C. S., Kim, J. E., Hong, Y. S., Kim, S. Y., Kim, T. W."Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance". Oncology Reports 47, no. 1 (2022): 15. https://doi.org/10.3892/or.2021.8226
Copy and paste a formatted citation
x
Spandidos Publications style
Tak E, Kim M, Cho Y, Choi S, Kim J, Han B, Kim H, Jang CS, Kim JE, Hong YS, Hong YS, et al: Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance. Oncol Rep 47: 15, 2022.
APA
Tak, E., Kim, M., Cho, Y., Choi, S., Kim, J., Han, B. ... Kim, T.W. (2022). Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance. Oncology Reports, 47, 15. https://doi.org/10.3892/or.2021.8226
MLA
Tak, E., Kim, M., Cho, Y., Choi, S., Kim, J., Han, B., Kim, H., Jang, C. S., Kim, J. E., Hong, Y. S., Kim, S. Y., Kim, T. W."Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance". Oncology Reports 47.1 (2022): 15.
Chicago
Tak, E., Kim, M., Cho, Y., Choi, S., Kim, J., Han, B., Kim, H., Jang, C. S., Kim, J. E., Hong, Y. S., Kim, S. Y., Kim, T. W."Expression of neurofibromin 1 in colorectal cancer and cetuximab resistance". Oncology Reports 47, no. 1 (2022): 15. https://doi.org/10.3892/or.2021.8226
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