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Article Open Access

Targeting the DNHD1‑HDAC6 axis inactivates prostate cancer cell aggressiveness in vitro, suggesting potential therapeutic vulnerability

  • Authors:
    • An Ni Lee
    • Chia Chang Wu
    • Su Wei Hu
    • Chih Heng Chen
    • Yi Te Chiang
    • Wei Tang Kao
    • Kai Yi Tzou
    • Chia Hung Liu
    • Shao Wei Dong
    • Yu Ting Wang
    • Yueh Lin Lee
    • Siou Bi Li
    • Che An Wu
    • Chiao Chun Liao
    • Ming Hsien Chan
    • Po Han Chiu
    • Chien Hsiu Li
  • View Affiliations / Copyright

    Affiliations: Department of Urology, Shuang Ho Hospital, Taipei Medical University, New Taipei City, Taiwan 23561, R.O.C., Department of Tropical Medicine, School of Medicine, College of Medicine, National Yang Ming Chiao Tung University, Taipei, Taiwan 300093, R.O.C., Department of Biomedical Imaging and Radiological Sciences, National Yang Ming Chiao Tung University, Taipei 112, Taiwan 300093, R.O.C.
    Copyright: © Lee et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY 4.0].
  • Article Number: 198
    |
    Published online on: September 30, 2026
       https://doi.org/10.3892/or.2026.9204
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Abstract

Prostate cancer progression toward an aggressive, therapy‑resistant disease state remains a major clinical challenge, indicating a need to identify novel molecular drivers of such progression and actionable therapeutic targets. The present study analyzed the role of dynein heavy chain domain 1 (DNHD1), a previously uncharacterized regulator of malignancy, in prostate cancer. Integrative transcriptomic analyses across The Cancer Genome Atlas and independent clinical cohorts revealed that DNHD1 is markedly upregulated in advanced‑stage prostate cancer and positively correlated with increasing Gleason scores. Elevated DNHD1 expression was associated with poor overall survival and disease‑free survival, which supports its clinical relevance as a prognostic biomarker. Functional analyses demonstrated that DNHD1 promoted tumor cell proliferation, colony formation, migration and invasion in vitro. Molecular correlation modeling and pathway analyses revealed histone deacetylase 6 (HDAC6) as a key downstream effector of DNHD1. DNHD1 expression was positively correlated with and regulated HDAC6 expression in clinical datasets. Gain‑of‑function and loss‑of‑function experiments further revealed that HDAC6 is required for DNHD1‑mediated oncogenic phenotypes in prostate cancer, positioning HDAC6 as a functional mediator downstream of DNHD1. The DNHD1‑HDAC6 axis was associated with cilium assembly‑related pathways, suggesting a previously unrecognized association between ciliary regulation and prostate cancer progression. HDAC6 inhibition with tubastatin A effectively suppressed DNHD1‑driven tumor growth and motility, as well as increased acetylated α‑tubulin and decreased androgen receptor signaling. These findings indicated a potential therapeutic vulnerability within the DNHD1‑HDAC6 axis. They implied that in prostate cancer, the DNHD1‑HDAC6 axis, a novel regulatory pathway, promotes aggressiveness and that HDAC6 is a targetable mediator of DNHD1‑driven malignancy. This axis can be considered a prognostic indicator and mechanistic foundation for developing targeted therapeutic strategies for advanced prostate cancer.
View Figures

Figure 1

Dysregulation of DNHD1
expression is associated with prostate cancer progression and
unfavorable clinical outcomes. (A) Differential expression analysis
of TCGA-PRAD transcriptomic profiles stratified by pathological
stage (T2a vs. T4), indicating DNHD1 as one of the
transcripts markedly enriched in advanced prostate cancer. (B)
DNHD1 expression levels in paired prostate tumor and
adjacent nontumor tissues from the TCGA-PRAD cohort, indicating
increased DNHD1 expression in tumor specimens. (C) Pancancer
expression profiling of DNHD1 across TCGA-PRAD cohorts,
indicating differential expression patterns among multiple human
cancer types. (D) Pancancer prognostic analyses of association
between DNHD1 expression and clinical outcome across
distinct tumor types, indicating that PRAD, KIRC and COAD are tumor
entities with significant DNHD1-associated prognostic
stratification. (E) Αnalysis between DNHD1 expression and
Gleason score in prostate cancer specimens. (F) Distribution of
DNHD1 expression across stratified Gleason grade score in
the TCGA-PRAD cohort. (G) Validation of DNHD1 expression
patterns in independent prostate cancer cohorts (GSE21032, GSE35988
and GSE48403), including benign prostate tissues as well as
localized prostate cancer, mCRPC and ADT-associated prostate cancer
tissues. (H) Kaplan-Meier survival analyses of OS and DFS according
to DNHD1 expression status in the TCGA-PRAD cohort. (I)
Independent validation of prognostic significance of DNHD1
expression for DFS in GSE21032. *P<0.05, **P<0.01,
***P<0.001. DNHD1, dynein heavy chain domain 1; TCGA, The Cancer
Genome Atlas; TPM, transcripts per million; mCRPC, metastatic
castration-resistant prostate cancer; ADT, androgen deprivation
therapy; OS, overall survival; DFS, disease-free survival; HR,
hazard ratio; GEO, Gene Expression Omnibus; ACC, Adrenocortical
Carcinoma; BLCA, Bladder Urothelial Carcinoma; BRCA, Breast
Invasive Carcinoma; CESC, Cervical Squamous Cell Carcinoma and
Endocervical Adenocarcinoma; CHOL, Cholangiocarcinoma; COAD, Colon
Adenocarcinoma; DLBC, Lymphoid Neoplasm Diffuse Large B-cell
Lymphoma; ESCA, Esophageal Carcinoma; GBM, Glioblastoma Multiforme;
HNSC, Head and Neck Squamous Cell Carcinoma; KICH, Kidney
Chromophobe; KIRC, Kidney Renal Clear Cell Carcinoma; KIRP, Kidney
Renal Papillary Cell Carcinoma; LAML, Acute Myeloid Leukemia; LGG,
Brain Lower Grade Glioma; LIHC, Liver Hepatocellular Carcinoma;
LUAD, Lung Adenocarcinoma; LUSC, Lung Squamous Cell Carcinoma;
MESO, Mesothelioma; OV, Ovarian Serous Cystadenocarcinoma; PAAD,
Pancreatic Adenocarcinoma; PCPG, Pheochromocytoma and
Paraganglioma; PRAD, Prostate Adenocarcinoma; READ, Rectum
Adenocarcinoma; SARC, Sarcoma; SKCM, Skin Cutaneous Melanoma; STAD,
Stomach Adenocarcinoma; TGCT, Testicular Germ Cell Tumors; THCA,
Thyroid Carcinoma; THYM, Thymoma; UCEC, Uterine Corpus Endometrial
Carcinoma; UCS, Uterine Carcinosarcoma; UVM, Uveal Melanoma.

Figure 2

DNHD1 facilitates prostate
cancer cell growth and metastatic behavior. (A) Analysis of
DNHD1 expression across prostate cancer cell lines from the
CCLE dataset (GSE36133), including AR-positive (LNCaP and 22RV1)
and AR-negative (PC-3 and DU145) cell models. (B) Immunoblotting of
DNHD1 expression in a panel of prostate cancer cell lines. (C and
D) Establishment and validation of DNHD1 gain-of-function
and loss-of-function models. DNHD1 overexpression in LNCaP
cells and DNHD1 knockdown in PC-3 cells were confirmed
through qPCR and immunoblotting. (E) Alamar Blue proliferation
assays indicating effects of DNHD1 overexpression in LNCaP
cells and DNHD1 knockdown in PC-3 cells on cell growth. (F)
Colony formation assays assessing clonogenic capacity after
DNHD1 overexpression in LNCaP cells or DNHD1
knockdown in PC-3 cells. (G) Boyden chamber migration assays for
migratory activity in DNHD1-overexpressing LNCaP cells and
DNHD1-knockdown PC-3 cells. (H) Matrigel invasion assays for
invasive capacity of DNHD1-overexpressing LNCaP cells and
DNHD1-knockdown PC-3 cells. (Scale bar, 100 µm). *P<0.05,
**P<0.01, ***P<0.001. DNHD1, dynein heavy chain domain 1;
CCLE, Cancer Cell Line Encyclopedia; AR, androgen receptor; qPCR,
quantitative polymerase chain reaction; shRNA, short hairpin RNA;
sh-Luc, short hairpin RNA targeting luciferase (control); Luc,
luciferase; AR-V7, androgen receptor splice variant 7; GSE, Gene
Expression Omnibus Series.

Figure 3

HDAC6 is a key downstream
effector within the DNHD1-associated oncogenic network in
prostate cancer. (A) Integrative correlation analysis across the
TCGA-PRAD, Cell 2015, Firehose Legacy and PanCancer Atlas cohorts
to identify genes consistently associated with DNHD1
expression (Spearman's ρ≥0.3 or Spearman's ρ≤-0.3), generating a
high-confidence DNHD1-associated transcriptional network.
(B) IPA of DNHD1-associated genes revealing significant
enrichment of biological processes related to cytoskeletal
organization, vesicular transport, organelle dynamics and metabolic
regulation. Cilium assembly appeared to be one of the most
consistently enriched pathways, implicating DNHD1 in
structural signaling programs associated with prostate cancer
progression. (C and D) Transcriptomic analyses of TCGA-PRAD
specimens demonstrating increased HDAC6 expression in
prostate cancer tissues relative to normal prostate tissues.
Representative comparisons and quantitative analyses are presented.
(E) Correlation analysis in the TCGA-PRAD cohort. The significant
positive association between DNHD1 and HDAC6
expression confirmed a coordinated transcriptional relationship
between these genes in clinical prostate cancer specimens. (F)
Clinicopathological analysis revealing a positive association
between HDAC6 expression and Gleason score, thereby
indicating progressive upregulation of HDAC6 expression with
increasing histological aggressiveness. (G) Independent validation
within prostate cancer patient cohorts, confirming a significant
positive correlation between DNHD1 and HDAC6
expression levels, supporting the robustness of the DNHD1-HDAC6
axis. (H) Validation across independent clinical datasets,
demonstrating elevated HDAC6 expression in localized
prostate cancer, mCRPC and ADT-treated prostate cancer tissues,
indicating its association with disease progression and
therapy-adaptive tumor states. (I) Kaplan-Meier survival analyses
demonstrating that upregulated HDAC6 expression is
associated with inferior OS and DFS in patients with prostate
cancer, supporting its prognostic relevance. (J) Combined
stratification based on DNHD1 and HDAC6 expression. A
high-risk molecular subgroup with the poorest clinical outcome was
noted, confirming the additive prognostic value of the DNHD1-HDAC6
axis. HDAC6, histone deacetylase 6; DNHD1, dynein heavy chain
domain 1; TCGA, The Cancer Genome Atlas; PRAD, prostate
adenocarcinoma; IPA, Ingenuity Pathway Analysis; COPII, coat
protein complex II; ER, endoplasmic reticulum; TPM, transcripts per
million; mCRPC, metastatic castration-resistant prostate cancer;
ADT, androgen deprivation therapy; OS, overall survival; DFS,
disease-free survival; HR, hazard ratio; GEO, Gene Expression
Omnibus; GSE, Gene Expression Omnibus Series.

Figure 4

DNHD1 promotes malignancy in
prostate cancer cells through HDAC6-dependent regulatory
mechanisms. (A) Analysis of the CCLE-derived prostate cancer
transcriptomic dataset GSE36133, demonstrating a lineage-associated
distribution pattern of HDAC6 expression, with lower
expression observed in AR-positive luminal-like prostate cancer
cell lines relative to AR-negative cell lines. (B) Correlation
analysis across prostate cancer datasets, revealing a strong
positive association between DNHD1 and HDAC6
expression (Spearman's ρ=0.90), confirming a coordinated
transcriptional relationship between these two molecules. (C and D)
Positive regulatory effect of DNHD1 on HDAC6
transcript and protein expression. Ectopic expression of
DNHD1 in LNCaP cells increased HDAC6 expression,
whereas DNHD1 knockdown in PC-3 cells reduced HDAC6
mRNA and protein expression, as determined through qPCR and
immunoblotting. (E) Reciprocal perturbation analyses demonstrating
that modulating HDAC6 expression did not alter DNHD1
expression, indicating a unidirectional regulatory relationship in
which DNHD1 functions as an upstream regulator of
HDAC6. (F and G) Establishment and validation of
HDAC6 gain-of-function and loss-of-function models in
prostate cancer cells. HDAC6 overexpression in LNCaP cells
and HDAC6 knockdown in PC-3 cells were confirmed through
qPCR and immunoblotting. Functional analyses demonstrating that
HDAC6 contributes to prostate cancer cell growth and
clonogenic capacity. HDAC6 overexpression (H) enhanced
proliferative and (I) colony-forming ability in LNCaP cells,
whereas HDAC6 knockdown suppressed these phenotypes in PC-3
cells. (J and K) Boyden chamber migration and Matrigel invasion
assays demonstrating that HDAC6 positively regulates
prostate cancer cell motility and invasive behavior. Ectopic
HDAC6 expression increased migratory and invasive capacities
in LNCaP cells, whereas HDAC6 knockdown markedly impaired
these phenotypes in PC-3 cells. (L-O) Epistasis analyses in
DNHD1-overexpressing LNCaP cells indicating that
HDAC6 is required for DNHD1-driven oncogenic phenotypes.
DNHD1 promoted cell proliferation, clonogenic growth,
migration and invasion, whereas concurrent HDAC6 silencing
markedly attenuated these tumor-promoting effects (Scale bar, 100
µm). *P<0.05, **P<0.01, ***P<0.001. DNHD1, dynein heavy
chain domain 1; HDAC6, histone deacetylase 6; CCLE, Cancer Cell
Line Encyclopedia; AR, androgen receptor; AR-V7, androgen receptor
splice variant 7; TPM, transcripts per million; qPCR, quantitative
polymerase chain reaction; mRNA, messenger RNA; shRNA, short
hairpin RNA; sh-Luc, short hairpin RNA targeting luciferase
(control); Luc, luciferase; shHDAC6, short hairpin RNA targeting
histone deacetylase 6; GSE, Gene Expression Omnibus Series.

Figure 5

Pharmacological inhibition of
HDAC6 suppresses DNHD1-driven oncogenic phenotypes in
prostate cancer. (A) Immunoblotting of LNCaP cells treated with
increasing concentrations of the selective HDAC6 inhibitor
tubastatin A. The results indicate dose-dependent accumulation of
acetylated α-tubulin, confirming inhibition of HDAC6 deacetylase
activity. (B) Time-course analysis in LNCaP cells demonstrating
sustained increase in acetylated α-tubulin levels after tubastatin
A treatment, thereby indicating prolonged suppression of
HDAC6-dependent deacetylation. (C) Pharmacological
inhibition of HDAC6. The results demonstrate reduced AR and
PSA expression in LNCaP cells, consistent with suppression of AR
signaling activity. (D-G) Functional analyses in LNCaP cells
demonstrating that tubastatin A treatment markedly inhibited cell
viability, clonogenic growth, migratory capacity and invasive
behavior. (H) Immunoblotting in DNHD1-overexpressing LNCaP
cells. The results indicate that tubastatin A treatment suppressed
DNHD1-associated HDAC6 expression and concomitantly
reduced AR and PSA expression. (I-L) Functional epistasis analyses
in DNHD1-overexpressing LNCaP cells. The results
demonstrated that pharmacological inhibition of HDAC6 markedly
attenuates DNHD1-induced enhancements in cell proliferation,
colony formation, migration and invasion, indicating that HDAC6
activity is required for DNHD1-mediated oncogenic effects in
prostate cancer. (Scale bar, 100 µm). *P<0.05, **P<0.01,
***P<0.001. DNHD1, dynein heavy chain domain 1; HDAC6, histone
deacetylase 6; Ac-α-tubulin, acetylated α-tubulin; SAHA,
suberoylanilide hydroxamic acid (vorinostat); AR, androgen
receptor; PSA, prostate-specific antigen; DMSO, dimethyl sulfoxide;
Luc, luciferase; sh-Luc, short hairpin RNA targeting luciferase
(control); shHDAC6, short hairpin RNA targeting histone deacetylase
6.

Figure 6

The DNHD1-HDAC6 axis is a
transcriptionally coordinated program associated with prostate
cancer progression and adverse clinical outcomes. Correlation
analyses of the relationship between DNHD1 and HDAC6
expression across distinct clinicopathological contexts within the
TCGA-PRAD cohort. Positive associations between DNHD1 and
HDAC6 expression were observed in tumors stratified by
pathological (A) N and (B) T stages and (C) residual tumor status,
confirming the maintenance of the regulatory relationship between
DNHD1 and HDAC6 during disease progression and
posttreatment prostate cancer states. Kaplan-Meier analyses of DFI
by (D) DNHD1 and (E) HDAC6 expression in patients
from the TCGA-PRAD cohort. Patients were stratified into high- and
low-expression groups on the basis of predefined cutoff criteria.
(F) Integrated prognostic stratification of DFI based on combined
DNHD1 and HDAC6 expression status. Coexpression
analysis revealed a patient subgroup exhibiting the most
unfavorable clinical outcomes. Kaplan-Meier analyses of PFI
according to (G) DNHD1 or (H) HDAC6 expression levels
in the TCGA-PRAD cohort. (I) Combined PFI analysis based on
DNHD1-HDAC6 coexpression status demonstrating enhanced prognostic
discrimination compared with single-gene stratification. (J)
Venn-based integration of DNHD1- and HDAC6-associated
transcriptional networks indicating a shared set of positively
correlated genes that represent the common downstream regulatory
program of the DNHD1-HDAC6 axis. (K) Functional enrichment analysis
of the shared gene set associated with the DNHD1-HDAC6 axis. After
exclusion of non-protein-coding transcripts, a refined 570-gene
signature was established, with cilium assembly emerging as the
most markedly enriched biological process. (L) Correlation analyses
demonstrating significant positive associations between the
DNHD1-HDAC6 downstream transcriptional signature and expression
levels of DNHD1 or HDAC6 in PRAD specimens,
confirming coordinated transcriptional coupling of the
axis-associated gene program. (M) Kaplan-Meier analyses of OS and
DFS according to the DNHD1-HDAC6 downstream transcriptional
signature. Patients were stratified into high- and low-signature
groups on the basis of predefined cutoff criteria, demonstrating
the prognostic relevance of the shared DNHD1-HDAC6 gene program.
DNHD1, dynein heavy chain domain 1; HDAC6, histone deacetylase 6;
TCGA, The Cancer Genome Atlas; PRAD, prostate adenocarcinoma; TPM,
transcripts per million; DFI, disease-free interval; PFI,
progression-free interval; OS, overall survival; DFS, disease-free
survival; HR, hazard ratio.
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Copy and paste a formatted citation
Spandidos Publications style
Lee AN, Wu CC, Hu SW, Chen CH, Chiang YT, Kao WT, Tzou KY, Liu CH, Dong SW, Wang YT, Wang YT, et al: Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability. Oncol Rep 56: 198, 2026.
APA
Lee, A.N., Wu, C.C., Hu, S.W., Chen, C.H., Chiang, Y.T., Kao, W.T. ... Li, C.H. (2026). Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability. Oncology Reports, 56, 198. https://doi.org/10.3892/or.2026.9204
MLA
Lee, A. N., Wu, C. C., Hu, S. W., Chen, C. H., Chiang, Y. T., Kao, W. T., Tzou, K. Y., Liu, C. H., Dong, S. W., Wang, Y. T., Lee, Y. L., Li, S. B., Wu, C. A., Liao, C. C., Chan, M. H., Chiu, P. H., Li, C. H."Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability". Oncology Reports 56.5 (2026): 198.
Chicago
Lee, A. N., Wu, C. C., Hu, S. W., Chen, C. H., Chiang, Y. T., Kao, W. T., Tzou, K. Y., Liu, C. H., Dong, S. W., Wang, Y. T., Lee, Y. L., Li, S. B., Wu, C. A., Liao, C. C., Chan, M. H., Chiu, P. H., Li, C. H."Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability". Oncology Reports 56, no. 5 (2026): 198. https://doi.org/10.3892/or.2026.9204
Copy and paste a formatted citation
x
Spandidos Publications style
Lee AN, Wu CC, Hu SW, Chen CH, Chiang YT, Kao WT, Tzou KY, Liu CH, Dong SW, Wang YT, Wang YT, et al: Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability. Oncol Rep 56: 198, 2026.
APA
Lee, A.N., Wu, C.C., Hu, S.W., Chen, C.H., Chiang, Y.T., Kao, W.T. ... Li, C.H. (2026). Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability. Oncology Reports, 56, 198. https://doi.org/10.3892/or.2026.9204
MLA
Lee, A. N., Wu, C. C., Hu, S. W., Chen, C. H., Chiang, Y. T., Kao, W. T., Tzou, K. Y., Liu, C. H., Dong, S. W., Wang, Y. T., Lee, Y. L., Li, S. B., Wu, C. A., Liao, C. C., Chan, M. H., Chiu, P. H., Li, C. H."Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability". Oncology Reports 56.5 (2026): 198.
Chicago
Lee, A. N., Wu, C. C., Hu, S. W., Chen, C. H., Chiang, Y. T., Kao, W. T., Tzou, K. Y., Liu, C. H., Dong, S. W., Wang, Y. T., Lee, Y. L., Li, S. B., Wu, C. A., Liao, C. C., Chan, M. H., Chiu, P. H., Li, C. H."Targeting the DNHD1‑HDAC6 axis inactivates prostate&nbsp;cancer cell aggressiveness <em>in vitro</em>, suggesting potential therapeutic vulnerability". Oncology Reports 56, no. 5 (2026): 198. https://doi.org/10.3892/or.2026.9204
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