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Article Open Access

PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming

  • Authors:
    • Zong Jin Guo
    • Qian Yu
    • Rui Sha
    • Wei Li
    • Hui Juan Dai
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    Affiliations: Division of Interventional Radiology, The University of Hong Kong‑Shenzhen Hospital, Shenzhen, Guangdong 518053, P.R. China, Department of Breast Surgery, Huai'an Maternal and Child Healthcare Center, Huai'an, Jiangsu 223002, P.R. China, Department of Thyroid and Breast Surgery, The First Affiliated Hospital of Wannan Medical College (Yijishan Hospital of Wannan Medical College), Wuhu, Anhui 241001, P.R. China, Department of Oncology Radiotherapy, The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu 223002, P.R. China, Department of Breast Surgery, Renji Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200127, P.R. China
    Copyright: © Guo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 112
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    Published online on: April 14, 2026
       https://doi.org/10.3892/or.2026.9117
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Abstract

Breast cancer is a predominant cause of cancer‑related mortality among women, particularly aggressive subtypes such as triple‑negative breast cancer (TNBC), which currently lack effective targeted therapies. While PTEN‑induced kinase 1 (PINK1) is known for its role in maintaining mitochondrial homeostasis via mitophagy, its specific contributions to breast cancer progression and metabolic regulation remain poorly defined. The present study aimed to investigate the oncogenic potential of PINK1 and its influence on metabolic reprogramming. To achieve this, the PINK1 expression levels in breast cancer tissues and cell lines were assessed. Gain‑ and loss‑of‑function methodologies were employed in luminal (MCF‑7) and TNBC (MDA‑MB‑231) cells. Then, mitophagy was evaluated by measuring LC3‑II levels, Parkin expression and utilizing transmission electron microscopy. Glucose uptake assays and metabolite quantification (including pyruvate and acetyl‑CoA) were conducted. Reverse transcription‑quantitative polymerase chain reaction identified phosphoglycerate kinase 2 (PGK2) as a downstream target of PINK1. Functional assays were then performed to examine the proliferation, migration and invasion of cells with PINK1 overexpression. The results demonstrated that PINK1 overexpression increased mitophagy and induced a glycolytic phenotype, characterized by enhanced glucose uptake and elevated PGK2 levels. Elevated concentrations of pyruvate and acetyl‑CoA indicated increased metabolic flux. Functionally, PINK1 promoted proliferation, migration and invasion in both cell types. Knockdown of PGK2 reversed these effects, underscoring its critical role in PINK1‑mediated metabolic reprogramming. Transcriptomic data obtained from online databases revealed a correlation between high PINK1 expression and immunosuppressive tumor microenvironments, as well as poor prognosis. The PINK1‑PGK2 axis constitutes a critical mechanism linking mitophagy to glycolytic reprogramming in breast cancer, representing a novel therapeutic target, particularly for TNBC. Targeting this axis may yield new strategies for addressing treatment‑resistant, metabolically adaptive breast cancer.
View Figures

Figure 1

Evaluation of mitophagy activity,
prognostic relevance and immune correlation of PINK1 in breast
cancer. Violin plots depicting enrichment scores of the (A)
GOBP_MITOPHAGY, (B) GOBP_REGULATION_OF_MITOPHAGY, (C)
REACTOME_PINK1_PARKIN_MEDIATED_MITOPHAGY and (D)
REACTOME_RECEPTOR_MEDIATED_MITOPHAGY gene sets in tumor and normal
tissues from TCGA-BRCA (Mann-Whitney U test). Prognostic analysis
of mitochondrial autophagy pathway enrichment scores of the (E)
GOBP_REGULATION_OF_MITOPHAGY and (F)
GOBP_POSITIVE_REGULATION_OF_MITOPHAGY gene sets. (G) Clinical
prognosis analysis of PINK1 (log-rank test). (H) Correlation
analysis between PINK1 and the immune cell infiltration scores of
22 types calculated by CIBERSORT (Spearman's rank correlation
analysis). Data are represented as the mean ± SEM from at least
three independent experiments. ****P<0.0001. PINK1, PTEN-induced
kinase 1; TCGA, The Cancer Genome Atlas; BRCA, Breast Invasive
Carcinoma; ns, not significant.

Figure 2

Analysis of PINK1 overexpression and
its impact on mitophagy markers in breast cancer cells. (A) RT-qPCR
analysis was used to measure PINK1 mRNA expression in MCF-7 cells
transfected with either a control vector or a PINK1 overexpression
plasmid. (B) Western blot analysis was performed to assess the
protein levels of PINK1, Parkin and LC3-I/II in MCF-7 cells. (C)
RT-qPCR analysis was used to measure PINK1 mRNA expression in
MDA-MB-231 cells transfected with either a control vector or a
PINK1 overexpression plasmid. (D) Western blot analysis was
performed to assess the protein levels of PINK1, Parkin and
LC3-I/II in MDA-MB-231 cells. Densitometric quantification of the
indicated proteins, normalized to β-actin, is shown for (E) MCF-7
and (F) MDA-MB-231 cells. The β-actin band labeled as ‘LC3’ denotes
the loading control for the LC3 experimental condition and was
separated on 15% SDS-PAGE; all other samples were separated on 10%
SDS-PAGE. Statistical analysis was performed using a unpaired
two-tailed Student's t-test. Data are represented as the mean ± SEM
from at least three independent experiments. *P<0.05,
***P<0.001. RT-qPCR, reverse transcription-quantitative PCR;
PINK1, PTEN-induced kinase 1; OE, overexpression.

Figure 3

Ultrastructural analysis of
PINK1-regulated mitophagy in breast cancer cells. TEM images of
MCF-7 cells transfected with either (A) vector control or (B) PINK1
overexpression plasmid. TEM images of MDA-MB-231 cells with (C)
vector control or (D) PINK1 overexpression plasmid. The yellow
arrows indicate autophagic structures formed during mitophagy.
PINK1, PTEN-induced kinase 1; TEM, transmission electron
microscopy; OE, overexpression.

Figure 4

Functional assessment of PINK1
overexpression in breast cancer cell proliferation and migration.
(A) Cell viability curve of MCF-7 cells transfected with either
vector control or PINK1 overexpression plasmid over 96 h. (B)
Quantitative analysis of MCF-7 cell viability at 96 h. (C) Cell
viability curve of MDA-MB-231 cells under the same conditions. (D)
Quantitative analysis of MDA-MB-231 viability. (E) Wound healing
images and (F) quantification of the migration ability of MCF-7
cells at 0 and 24 h. (G) Wound healing images and quantification of
(H) the migration ability of MDA-MB-231 cells at 0 and 24 h.
Statistical analysis was performed using a unpaired two-tailed
Student's t-test. Data are represented as the mean ± SEM from at
least three independent experiments. **P<0.01, ***P<0.001.
PINK1, PTEN-induced kinase 1; OE, overexpression.

Figure 5

Transwell invasion assay illustrating
the effect of PINK1 overexpression on breast cancer cell
invasiveness. (A) Representative Transwell invasion images and (B)
quantification of the number of invasive MCF-7 cells transfected
with control vector or PINK1 overexpression plasmid. (C)
Representative Transwell invasion images and (D) quantification of
the number of invasive MDA-MB-231 cells transfected with control
vector or PINK1 overexpression plasmid. Statistical analysis was
performed using a unpaired two-tailed Student's t-test. Data
represent means ± SEM from at least three independent experiments.
**P<0.01, ***P<0.001. PINK1, PTEN-induced kinase 1; OE,
overexpression.

Figure 6

Analysis of PINK1 knockdown and its
impact on mitophagy-related proteins in breast cancer cells. (A)
Relative mRNA levels of PINK1 in MCF-7 cells transfected with shRNA
targeting PINK1 or a negative control were measured by quantitative
PCR. (B) Western blot analysis displayed PINK1, Parkin and LC3-I/II
protein levels in MCF-7 cells following PINK1 knockdown. (C)
Relative mRNA levels of PINK1 in MDA-MB-231 cells transfected with
shRNA targeting PINK1 or a negative control were measured by
quantitative PCR. (D) Western blot analysis displayed PINK1, Parkin
and LC3-I/II protein levels in MDA-MB-231 cells following PINK1
knockdown. Densitometric quantification of PINK1, Parkin and LC3-II
protein expression in (E) MCF-7 and (F) MDA-MB-231 cells is also
presented. The β-actin band labeled as ‘LC3’ denotes the loading
control for the LC3 experimental condition and was separated on 15%
SDS-PAGE; all other samples were separated on 10% SDS–PAGE.
Statistical analysis was performed using a unpaired two-tailed
Student's t-test. Data are represented as the mean ± SEM from at
least three independent experiments. *P<0.05, **P<0.01,
***P<0.001. PINK1, PTEN-induced kinase 1; sh, short hairpin; NC,
negative control.

Figure 7

Ultrastructural assessment of
PINK1-induced mitophagy in breast cancer cells. TEM images of MCF-7
cells transfected with either (A) sh-NC or (B) sh-PINK1 groups. TEM
images of MDA-MB-231 cells transfected with either (C) sh-NC or (D)
sh-PINK1 groups. The yellow arrows indicate autophagic structures
formed during mitophagy. PINK1, PTEN-induced kinase 1; TEM,
transmission electron microscopy; sh, short hairpin; NC, negative
control.

Figure 8

Functional assessment of PINK1
knockdown on breast cancer cell proliferation and migration. (A)
Cell viability curves and (B) 96 h statistical comparison of MCF-7
cells transfected with shRNA targeting PINK1 or a negative control.
(C) Cell viability curves and (D) 96 h statistical comparison of
MDA-MB-231 cells transfected with shRNA targeting PINK1 or a
negative control. (E) Representative wound healing images and (F)
corresponding migration quantification of the migratory capacity of
MCF-7 cells after PINK1 knockdown at 0 and 24 h. (G) Representative
wound healing images and (H) quantification of the migratory
capacity of MDA-MB-231 cells at 0 and 24 h. Statistical analysis
was performed using an unpaired two-tailed Student's t-test for
pairwise comparisons. Data are presented as the mean ± SEM from at
least three independent experiments. **P<0.01, ***P<0.001.
PINK1, PTEN-induced kinase 1; sh, short hairpin; NC, negative
control.

Figure 9

Transwell invasion assay evaluating
the impact of PINK1 knockdown on breast cancer cell invasiveness.
(A) Representative Transwell invasion images and (B) quantitative
analysis of the number of invasive MCF-7 cells transfected with
shRNA targeting PINK1 or negative control. (C) Representative
Transwell invasion images and (D) quantitative analysis of the
number of invasive MDA-MB-231 cells transfected with shRNA
targeting PINK1 or negative control. Statistical analysis was
performed using a unpaired two-tailed Student's t-test. Data are
represented as the mean ± SEM from at least three independent
experiments. **P<0.01, ***P<0.001. PINK1, PTEN-induced kinase
1; sh, short hairpin; NC, negative control.

Figure 10

Correlation analysis of
PINK1-mediated mitophagy with glycolysis and patient prognosis in
breast cancer. (A) Intersection analysis between DEGs and
Glyco-DEGs in the TCGA-BRCA dataset. (B) GSEA analysis of the
glycolysis pathway in the TCGA-BRCA dataset. (C) Prognostic
analysis of the glycolysis pathway activity in the TCGA-BRCA
cohort. (D) Correlation between the glycolysis score and the
activity of the REACTOME_PINK1_PRKN_MEDIATED_MITOPHAGY pathway. (E)
Correlation between the PINK1-mediated mitophagy pathway activity
and hallmark glycolysis gene set activity. (F) Correlation between
the PINK1-mediated mitophagy pathway activity and glycolysis core
signature gene set activity. (G) Correlation analysis between the
PINK1-mediated mitophagy pathway and glycolysis differential genes.
(H) Western blot analysis of PGK2 protein expression in MCF-7 and
MDA-MB-231 cells following PINK1 overexpression or knockdown.
Quantification of PGK2 levels is shown on the right, normalized to
β-actin. Statistical analyses were performed using GSEA, log-rank
test and Spearman correlation for the bioinformatics data, and
unpaired two-tailed Student's t-test for experimental validation.
Data represent means ± SEM from at least three independent
experiments. **P<0.01, ****P<0.0001. PINK1, PTEN-induced
kinase 1; DEGs, differentially expressed genes; Glyco, glycolysis;
TCGA, The Cancer Genome Atlas; BRCA, Breast Invasive Carcinoma;
GSEA, Gene Set Enrichment Analysis; NES, Normalized Enrichment
Score; PGK2, Phosphoglycerate Kinase 2.

Figure 11

Flow cytometric analysis of glucose
uptake in breast cancer cells upon PINK1 overexpression and PGK2
knockdown. (A) Flow cytometric analysis of glucose uptake in MCF-7
and MDA-MB-231 cells transfected with vector control, PINK1-OE or
PINK1-OE + siPGK2. Quantitative comparison of glucose-positive cell
populations in (B) MCF-7 and (C) MDA-MB-231 cells. Statistical
analysis was performed using one-way ANOVA followed by Tukey's
post-hoc test. Data are represented as the mean ± SEM from at least
three independent experiments. *P<0.05, ***P<0.001. PINK1,
PTEN-induced kinase 1; OE, overexpression; si, small interfering;
PGK2, phosphoglycerate kinase 2.

Figure 12

Wound healing assay evaluating the
effect of PINK1 overexpression and PGK2 knockdown on cell migration
in breast cancer cells. (A) Representative images of wound healing
in MCF-7 cells transfected with vector control, PINK1-OE or
PINK1-OE + siPGK2 groups at 0 and 24 h. (B) Quantitative analysis
of the relative migration distance in MCF-7 cells. (C)
Representative images of wound healing in MDA-MB-231 cells
transfected with vector control, PINK1-OE or PINK1-OE + siPGK2
groups at 0 and 24 h. (D) Quantitative analysis of the relative
migration distance in MDA-MB-231 cells. Statistical analysis was
performed using one-way ANOVA followed by Tukey's post-hoc test.
Data are represented as the mean ± SEM from at least three
independent experiments. *P<0.05, **P<0.01, ***P<0.001.
PINK1, PTEN-induced kinase 1; OE, overexpression; si, small
interfering; PGK2, phosphoglycerate kinase 2.

Figure 13

Mechanistic analysis of the
PINK1-PGK2 axis in regulating breast cancer invasion through
metabolic reprogramming. (A) Representative images of Transwell
invasion assays with MCF-7 cells and MDA-MB-231 cells transfected
with vector control, PINK1-OE or PINK1-OE + siPGK2. Quantitative
analysis of invasive cell numbers under the indicated conditions in
MCF-7 (B) cells and MDA-MB-231 (C) cells. Statistical analysis was
performed using a one-way ANOVA followed by Tukey's post-hoc test.
Data are represented as the mean ± SEM from at least three
independent experiments. **P<0.01, ***P<0.001,
****P<0.0001. PINK1, PTEN-induced kinase 1; OE, overexpression;
si, small interfering; PGK2, phosphoglycerate kinase 2.

Figure 14

Analysis of glycolysis-associated
metabolite level changes in MCF-7 and MDA-MB-231 breast cancer
cells. Levels of (A) PGK2, (B) pyruvate and (C) acetyl-CoA in MCF-7
cells, and (D) PGK2, (E) pyruvate and (F) acetyl-CoA in MDA-MB-231
cells transfected with vector control, PINK1-OE or PINK1-OE +
siPGK2. Quantitative analysis was performed using ELISA in cells
under the three indicated conditions. Statistical analysis was
performed using a one-way ANOVA followed by Tukey's post-hoc test.
Data are represented as the mean ± SEM from at least three
independent experiments. ***P<0.001. PGK2, phosphoglycerate
kinase 2; PINK1, PTEN-induced kinase 1; OE, overexpression; si,
small interfering.
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Copy and paste a formatted citation
Spandidos Publications style
Guo ZJ, Yu Q, Sha R, Li W and Dai HJ: PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming. Oncol Rep 55: 112, 2026.
APA
Guo, Z.J., Yu, Q., Sha, R., Li, W., & Dai, H.J. (2026). PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming. Oncology Reports, 55, 112. https://doi.org/10.3892/or.2026.9117
MLA
Guo, Z. J., Yu, Q., Sha, R., Li, W., Dai, H. J."PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming". Oncology Reports 55.6 (2026): 112.
Chicago
Guo, Z. J., Yu, Q., Sha, R., Li, W., Dai, H. J."PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming". Oncology Reports 55, no. 6 (2026): 112. https://doi.org/10.3892/or.2026.9117
Copy and paste a formatted citation
x
Spandidos Publications style
Guo ZJ, Yu Q, Sha R, Li W and Dai HJ: PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming. Oncol Rep 55: 112, 2026.
APA
Guo, Z.J., Yu, Q., Sha, R., Li, W., & Dai, H.J. (2026). PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming. Oncology Reports, 55, 112. https://doi.org/10.3892/or.2026.9117
MLA
Guo, Z. J., Yu, Q., Sha, R., Li, W., Dai, H. J."PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming". Oncology Reports 55.6 (2026): 112.
Chicago
Guo, Z. J., Yu, Q., Sha, R., Li, W., Dai, H. J."PINK1‑mediated mitophagy enhances breast cancer proliferation through metabolic reprogramming". Oncology Reports 55, no. 6 (2026): 112. https://doi.org/10.3892/or.2026.9117
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