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Article

DHX15 overexpression suppresses colorectal cancer cell line proliferation

  • Authors:
    • Yuki Ii
    • Kosuke Saita
    • Toshiro Iwagawa
    • Shingo Ito
    • Kiichi Sugimoto
    • Kazuhiro Sakamoto
    • Sumiko Watanabe
  • View Affiliations / Copyright

    Affiliations: Department of Coloproctological Surgery, Juntendo University Graduate School of Medicine, Tokyo 113‑8421, Japan, Department of Retinal Biology and Pathology, Graduate School of Medicine, University of Tokyo, Tokyo 113‑8655, Japan, Department of Gastrointestinal Surgery, Shonan Kamakura General Hospital, Kanagawa 247‑0072, Japan
  • Article Number: 80
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    Published online on: December 18, 2025
       https://doi.org/10.3892/ol.2025.15433
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Abstract

DEAH (Asp‑Glu‑Ala‑His) box helicase 15 (DHX15) is a member of the DEAD box RNA helicase family that carries out a key role in innate immunity against viral infections. It is involved in tumorigenesis as a tumor‑promoting factor in various types of cancer, but it has also been suggested to act as a tumor suppressor. However, the role of DHX15 in colorectal cancer (CRC) remains largely unknown. In the present study, we examined the role of DHX15 in CRC. Immunostaining of clinical samples from patients with CRC identified DHX15 proteins in the cell nuclei of both tumor and adjacent normal tissues. DHX15 overexpression was revealed to reduce the cell number of various CRC cell lines, as well as the number of Ki67‑positive proliferating cells. However, the number of AC3‑positive apoptotic cells was comparable between the control and DHX15‑overexpressing cells. The possible downstream mechanisms of DHX15 were further examined which revealed that activation of the Wnt/β‑catenin and NF‑κB signaling pathways were not affected by DHX15 expression. However, DHX15 overexpression resulted in fewer LC3 puncta in HCT116 and DLD1 cells. Taken together, DHX15 may negatively affect CRC cell proliferation, and autophagy may potentially be involved in the downstream mechanism of DHX15.
View Figures

Figure 1

DHX15 immunohistochemistry of human
CRC clinical samples. (A) Representative images of tumor and
adjacent normal tissue. DHX15 expression in tumor and adjacent
non-tumor tissues was examined by immunohistochemistry. Evaluation
of expression strength was carried out using ImageJ and percentage
of positive signals are shown in the bottom of the panel. (B)
Average of DHX15 signal-positive nuclei population of normal and
tumor tissues of 10 samples. (C) Representative images of
examination of intensity of DHX15 staining signals. Left panels are
imuunostained patterns and right panels are converted to 8-bit
color index. (D) Signal intensity of adjacent normal tissues and
tumor tissues of the 10 samples. (E) Average DHX15 signal intensity
of normal and tumor tissues of 10 samples. Scale bars represent 100
µm in A and C. *P<0.01; **P<0.01. DHX15, DEAH
(Asp-Glu-Ala-His) box helicase 15.

Figure 2

Effects of DHX15 overexpression on
the proliferation of various CRC cell lines. The CRC cell lines
HCT116, SW480, Caco2 and DLD1 were transfected with
DHX15-expressing or empty vector with EGFP-expressing vector. The
cells were incubated in IncuCyte ZOOM, and live-cell images were
captured every 8 h. (A) The population of EGFP positive cells was
analyzed using IncuCyte Live-Cell Imaging Software. The captured
images of HCT116 of control and DHX15 overexpressed cells at 0, 16,
32 and 48 h after transfection. (B) Transition of number of EGFP
positive cells from 0 to 48 h after transfection of empty vector or
DHX15 overexpression vector. Cell proliferation was examined by
Ki67 immunostaining. Transfected cells were cultured for 24 h,
followed by immunostaining with an anti-Ki67 antibody. (C)
Representative images, DAPI (blue) shows nucleus. (D) Population of
Ki67 positive cells in total EGFP positive cells in the 4 cell
lines. Scale bar, 100 µm in A and C. Data are average of 3
independent samples with SEM. Statistical significance was
calculated by Student's t test (two tailed). *P<0.01;
**P<0.01. DHX15, DEAH (Asp-Glu-Ala-His) box helicase 15; CRC,
colorectal cancer.

Figure 3

Effects of DHX15 overexpression on
CRC cells line apoptosis. Apoptosis of the DHX15-overexpressing CRC
cell lines was examined using active Caspase-3 (AC3)
immunostaining. Transfected cells were cultured 24 h and
immunostaining with anti-AC3 antibody was carried out. DAPI (blue)
shows nucleus. (A) Representative images of each sample. (B)
Population of AC3 positive cells in total EGFP positive cells in
the 4 cell lines. Scale bar, 100 µm in A. Data are average of 3
independent samples with SEM. Statistical significance was
calculated by Student's t test (two tailed), all comparison were
P>0.05 (B). DHX15, DEAH (Asp-Glu-Ala-His) box helicase 15; CRC,
colorectal cancer.

Figure 4

Effects of DHX15 overexpression on
the activation of Wnt, NF-κB and autophagy. CRC cell lines were
transfected with DHX15 or vector control with luciferase reporter
plasmids to detect Wnt and NF-κB activities and then harvested
after 18 h of incubation. CRC cell lines were transfected with
DHX15 or vector control with EGFP-expression plasmids and then
immunostained with anti-LC3 antibody after 8 h of culture.
Luciferase activities of each (A) TCF-luc or (B) NF-κB-luc cell
line. (C) The number of LC3-positive puncta signals among all
EGFP-positive cell lines. Data are average of 3 independent samples
with SEM. Statistical significance was calculated by Student's t
test (two tailed). *P<0.01; **P<0.01. DHX15, DEAH
(Asp-Glu-Ala-His) box helicase 15; CRC, colorectal cancer.
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Copy and paste a formatted citation
Spandidos Publications style
Ii Y, Saita K, Iwagawa T, Ito S, Sugimoto K, Sakamoto K and Watanabe S: DHX15 overexpression suppresses colorectal cancer cell line proliferation. Oncol Lett 31: 80, 2026.
APA
Ii, Y., Saita, K., Iwagawa, T., Ito, S., Sugimoto, K., Sakamoto, K., & Watanabe, S. (2026). DHX15 overexpression suppresses colorectal cancer cell line proliferation. Oncology Letters, 31, 80. https://doi.org/10.3892/ol.2025.15433
MLA
Ii, Y., Saita, K., Iwagawa, T., Ito, S., Sugimoto, K., Sakamoto, K., Watanabe, S."DHX15 overexpression suppresses colorectal cancer cell line proliferation". Oncology Letters 31.2 (2026): 80.
Chicago
Ii, Y., Saita, K., Iwagawa, T., Ito, S., Sugimoto, K., Sakamoto, K., Watanabe, S."DHX15 overexpression suppresses colorectal cancer cell line proliferation". Oncology Letters 31, no. 2 (2026): 80. https://doi.org/10.3892/ol.2025.15433
Copy and paste a formatted citation
x
Spandidos Publications style
Ii Y, Saita K, Iwagawa T, Ito S, Sugimoto K, Sakamoto K and Watanabe S: DHX15 overexpression suppresses colorectal cancer cell line proliferation. Oncol Lett 31: 80, 2026.
APA
Ii, Y., Saita, K., Iwagawa, T., Ito, S., Sugimoto, K., Sakamoto, K., & Watanabe, S. (2026). DHX15 overexpression suppresses colorectal cancer cell line proliferation. Oncology Letters, 31, 80. https://doi.org/10.3892/ol.2025.15433
MLA
Ii, Y., Saita, K., Iwagawa, T., Ito, S., Sugimoto, K., Sakamoto, K., Watanabe, S."DHX15 overexpression suppresses colorectal cancer cell line proliferation". Oncology Letters 31.2 (2026): 80.
Chicago
Ii, Y., Saita, K., Iwagawa, T., Ito, S., Sugimoto, K., Sakamoto, K., Watanabe, S."DHX15 overexpression suppresses colorectal cancer cell line proliferation". Oncology Letters 31, no. 2 (2026): 80. https://doi.org/10.3892/ol.2025.15433
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