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c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression

  • Authors:
    • Yanqiu Meng
    • Lebin Yuan
    • Gangrui Meng
    • Hongxiang Huang
    • Xianbin Huang
    • Xinping Xu
    • Xiaodong Peng
  • View Affiliations / Copyright

    Affiliations: Department of Oncology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, P.R. China, Department of Thyroid and Breast Surgery, Affiliated Hospital of Hubei University of Arts and Science, Xiangyang Central Hospital, Xiangyang, Hubei 441000, P.R. China, Department of General Surgery, First Clinical College, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China, Department of Oncology, Ganzhou Hospital‑Nanfang Hospital, Southern Medical University, Ganzhou, Jiangxi 341000, P.R. China, Jiangxi Provincial Key Laboratory of Respiratory Diseases, Jiangxi Institute of Respiratory Diseases, The Department of Respiratory and Critical Care Medicine, Jiangxi Clinical Research Center for Respiratory Diseases, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, Jiangxi 330006, P.R. China
    Copyright: © Meng et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
  • Article Number: 13
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    Published online on: November 27, 2025
       https://doi.org/10.3892/ijo.2025.5826
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Abstract

Hepatocellular carcinoma (HCC) continues to rank as a predominant contributor to cancer‑related mortality on a global scale, attributed to its insidious onset and unfavorable prognosis. The ribosomal protein lateral stalk subunit P0 (RPLP0) has recently gathered widespread attention as a crucial factor in the pathological progression of various neoplasms; however, its exact role in HCC remains inadequately defined. Consequently, the present study endeavored to shed light on the function and mechanistic underpinnings of RPLP0 in HCC and assess its clinical significance and potential as a therapeutic target. qPCR and western blot analyses indicated that RPLP0 was markedly upregulated in HCC, with its elevated levels correlating with poorer survival outcomes. Silencing RPLP0 expression suppressed the proliferative, invasive, migratory, and epithelial‑mesenchymal transition (EMT) abilities of HCC cells, while concurrently promoting apoptosis, autophagy, and G2/M cell cycle arrest, as evidenced by CCK‑8, colony formation, Transwell assays and flow cytometry analysis, respectively. Moreover, the findings revealed that RPLP0 downregulation mediated the suppression of the JAK2/STAT3 pathway through reactive oxygen species (ROS) accumulation, which in turn downregulated c‑Myc expression. Furthermore, chromatin immunoprecipitation and dual luciferase assays demonstrated that c‑Myc directly bound to the promoter sequence of RPLP0, thereby augmenting its transcriptional activity. In summary, the current study highlighted that RPLP0 establishes a feedback circuit with c‑Myc by facilitating JAK2/STAT3 pathway activation through suppressing ROS levels, while c‑Myc reciprocally activates RPLP0, forming a regulatory circuit loop that drives HCC progression. Thus, targeting the c‑Myc/RPLP0/ROS/JAK2/STAT3 axis emerges as a promising therapeutic strategy for the management of HCC.
View Figures

Figure 1

Elevated RPLP0 expression is
associated with unfavorable survival outcomes in HCC. (A) RPLP0
expression levels derived from the TCGA_GTEx database. (B)
Confirmation of RPLP0 expression in the GSE57957, GSE36411,
GSE39791 and GSE76427 datasets. Association between RPLP0
expression status and (C) OS and (D) DFS among HCC patients in the
GEPIA2 database. The relationship between RPLP0 expression and (E)
histological grade and (F) cancer stage in the TCGA database. The
expression levels of (G) mRNA and (H) protein for RPLP0 were
analyzed in 44 paired HCC samples. (I) The influence of RPLP0
expression on the survival outcomes of HCC patients. (J)
Immunohistochemical analysis of RPLP0 in HCC and adjacent
non-cancerous tissues (scale bar, 100 μm). RPLP0 (K) mRNA
and (L) protein levels were examined in immortalized hepatocytes
and HCC cell lines. *P<0.05, **P<0.01,
***P<0.001. RPLP0, ribosomal protein lateral stalk
subunit P0; HCC, hepatocellular carcinoma; TCGA, the cancer genome
atlas; GTEx, the genotype-tissue expression; N, normal; T, tumor;
GEO, gene expression omnibus; GEPIA2, gene expression profiling
interactive analysis; OS, overall survival; DFS, disease-free
survival.

Figure 2

RPLP0 overexpression promotes
malignant phenotypes in HCC cells. Alterations in mRNA and protein
levels following (A and B) RPLP0 knockdown and (C and D)
overexpression in HCC cells. (E) CCK-8, (F) colony formation and
(G) Transwell assays were conducted to assess HCC cell
proliferation, invasion and migration (scale bar, 150 μm).
(H) EMT-related protein expression changes (E-cadherin, Vimentin
and Snail) were observed in treated HCC cells.
*P<0.05, **P<0.01,
***P<0.001. RPLP0, ribosomal protein lateral stalk
subunit P0; HCC, hepatocellular carcinoma; CCK-8, cell counting
kit-8; EMT, epithelial-mesenchymal transition.

Figure 3

RPLP0 downregulation triggers
apoptosis and G2/M cell cycle arrest in HCC cells. Flow
cytometry was performed to evaluate the (A) apoptosis rate and (B)
cell cycle. (C) Western blotting was performed to quantify
apoptosis- and cycle-related protein levels (C-PARP, PARP, Cyclin
D1 and Bcl-2). *P<0.05, **P<0.01,
***P<0.001. RPLP0, ribosomal protein lateral stalk
subunit P0; HCC, hepatocellular carcinoma; PARP, Poly (ADP-ribose)
polymerase; C-, cleaved.

Figure 4

RPLP0 knockdown induces autophagy in
HCC cells. (A) GSEA demonstrated a significant enrichment of low
RPLP0 expression during autophagy. (B) Alterations in the levels of
autophagy-related proteins (P62 and LC3B). (C) The presence of
autophagosomes was assessed using MDC staining in HCC cells
subjected to RPLP0 knockdown and overexpression (scale bar, 50
μm). (D) Autophagosome quantity was examined via
transmission electron microscopy, with red arrows denoting
autophagosomes (scale bar, 2 μm). *P<0.05,
**P<0.01, ***P<0.001. RPLP0, ribosomal
protein lateral stalk subunit P0; HCC, hepatocellular carcinoma;
GSEA, gene set enrichment analysis; MDC, monodansylcadaverine.

Figure 5

RPLP0 knockdown stimulates autophagic
flux in HCC cells, as shown by 3-MA and CQ. (A) Western blotting
analyzed LC3B expression in HCC cells pretreated with transfection
and subsequently exposed to 5 mM 3-MA for 0, 12 and 24 h. (B) HCC
cells were treated with 20 μM CQ for 0, 12 and 24 h,
respectively, followed by RPLP0 knockdown, and western blotting
evaluated LC3B levels. *P<0.05,
**P<0.01, ***P<0.001. RPLP0, ribosomal
protein lateral stalk subunit P0; HCC, hepatocellular carcinoma;
3-MA, 3-methyladenine; CQ, chloroquine.

Figure 6

Suppression of RPLP0 expression
impedes xenograft tumor growth. (A) Subcutaneous xenograft tumor
models were established following pretreatment of MHCC97-H cells.
(B) Xenograft tumors were successfully isolated. The (C) volume and
(D) weight of subcutaneous xenografts were compared between the
shNC and shRPLP0 groups. (E) Western blotting was performed to
assess RPLP0, Ki-67, E-cadherin, Vimentin, Snail and LC3B levels in
the xenografts. (F) Immunohistochemical staining for Ki-67 in the
shNC and shRPLP0 groups (scale bar, 50 μm).
*P<0.05, **P<0.01,
***P<0.001. RPLP0, ribosomal protein lateral stalk
subunit P0; sh, short hairpin.

Figure 7

Suppression of RPLP0 expression
promotes ROS accumulation. (A) GSEA demonstrated a significant
correlation between reduced RPLP0 expression and increased ROS
accumulation. (B) ROS levels were evaluated via flow cytometry
following pretreatment. (C) Fluorescence microscopy was utilized to
evaluate ROS levels (scale bar, 50 μm). (D) HCC cells were
treated with 5 mM NAC for 0, 12 and 24 h and LC3B expression was
analyzed via western blotting. *P<0.05,
**P<0.01, ***P<0.001. RPLP0, ribosomal
protein lateral stalk subunit P0; ROS, reactive oxygen species;
GSEA, gene set enrichment analysis; HCC, hepatocellular carcinoma;
NAC, N-acetyl-L-cysteine.

Figure 8

Suppressing RPLP0 deactivates the
JAK2/STAT3 pathway by accumulating ROS. (A) Alterations in RPLP0,
p-JAK2/JAK2 and p-STAT3/STAT3 expression were assessed in HCC cells
following transfection. (B) After 48 h of transfection, Huh7 and
MHCC97-H cells were exposed to 40 ng/ml IL-6 for an additional 24
h, followed by western blot analysis of RPLP0, p-JAK2/JAK2 and
p-STAT3/STAT3 expression. (C) HCC cells underwent a 48-h
transfection pretreatment and were then treated with 5 mM NAC for
0, 12 and 24 h, followed by western blot analysis of RPLP0,
p-JAK2/JAK2 and p-STAT3/STAT3 expression. *P<0.05,
**P<0.01, ***P<0.001. RPLP0, ribosomal
protein lateral stalk subunit P0; ROS, reactive oxygen species; p-,
phosphorylated; HCC, hepatocellular carcinoma; NAC,
N-acetyl-L-cysteine.

Figure 9

c-Myc promotes RPLP0 transcription.
(A) The Cistrome Data Browser database was used to predict
transcription factors associated with RPLP0. Correlation analyses
of RPLP0 and c-Myc expression were conducted from the (B) TCGA
database and (C) HCC samples (n=44). (D) HCC cells were transfected
for 24 h to evaluate the knockdown efficiency of c-Myc mRNA and (E)
the corresponding expression level of RPLP0 mRNA. (F) HCC cells
were transfected for 48 h to examine alterations in c-Myc and RPLP0
protein expression. (G) A schematic illustration of c-Myc binding
motifs, along with the relative scoring of binding sites derived
from the JASPAR database. Corresponding primers were designed based
on these binding sites. (H) ChIP-qPCR was conducted to investigate
the binding of c-Myc to RPLP0 promoter sequences. (I) Agarose gel
electrophoresis was performed to visualize the binding of c-Myc to
RPLP0 promoter sequences (input as a positive control, IgG as a
negative control). (J) Identification of potential sites for c-Myc
binding to the RPLP0 promoter region. (K) HCC cells were
co-transfected and subsequently analyzed for luciferase activity.
**P<0.01, ***P<0.001. RPLP0, ribosomal
protein lateral stalk subunit P0; HCC, hepatocellular carcinoma;
ChIP, chromatin immunoprecipitation; qPCR, quantitative polymerase
chain reaction.

Figure 10

RPLP0 facilitates the proliferation
and metastasis of HCC cells through the regulation of c-Myc. (A)
CCK-8, (B) colony formation and (C) Transwell assays were used to
assess cell proliferation, migration and invasion (scale bar, 150
μm). (D) EMT-related protein levels, such as E-cadherin,
Vimentin and Snail, were examined in co-transfected HCC cells.
*P<0.05, **P<0.01,
***P<0.001. RPLP0, ribosomal protein lateral stalk
subunit P0; HCC, hepatocellular carcinoma; EMT,
epithelial-mesenchymal transition.

Figure 11

RPLP0-mediated regulation of
apoptosis and cell cycle distribution in HCC cells depends on
c-Myc. The (A) apoptosis rate and (B) cell cycle distribution were
analyzed via flow cytometry. (C) Western blotting was used to
evaluate PARP, C-PARP, Cyclin D1, Bcl-2 and LC3B expression.
*P<0.05, **P<0.01,
***P<0.001. RPLP0, ribosomal protein lateral stalk
subunit P0; HCC, hepatocellular carcinoma; PARP, Poly (ADP-ribose)
polymerase; C-, cleaved.

Figure 12

RPLP0, regulated by c-Myc,
facilitates HCC progression by reducing ROS levels and activating
the JAK2/STAT3 signaling pathway. Functional model of the
c-Myc/RPLP0/ROS/JAK2/STAT3 signaling axis. RPLP0, ribosomal protein
lateral stalk subunit P0; HCC, hepatocellular carcinoma; ROS,
reactive oxygen species.
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Copy and paste a formatted citation
Spandidos Publications style
Meng Y, Yuan L, Meng G, Huang H, Huang X, Xu X and Peng X: c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression. Int J Oncol 68: 13, 2026.
APA
Meng, Y., Yuan, L., Meng, G., Huang, H., Huang, X., Xu, X., & Peng, X. (2026). c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression. International Journal of Oncology, 68, 13. https://doi.org/10.3892/ijo.2025.5826
MLA
Meng, Y., Yuan, L., Meng, G., Huang, H., Huang, X., Xu, X., Peng, X."c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression". International Journal of Oncology 68.1 (2026): 13.
Chicago
Meng, Y., Yuan, L., Meng, G., Huang, H., Huang, X., Xu, X., Peng, X."c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression". International Journal of Oncology 68, no. 1 (2026): 13. https://doi.org/10.3892/ijo.2025.5826
Copy and paste a formatted citation
x
Spandidos Publications style
Meng Y, Yuan L, Meng G, Huang H, Huang X, Xu X and Peng X: c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression. Int J Oncol 68: 13, 2026.
APA
Meng, Y., Yuan, L., Meng, G., Huang, H., Huang, X., Xu, X., & Peng, X. (2026). c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression. International Journal of Oncology, 68, 13. https://doi.org/10.3892/ijo.2025.5826
MLA
Meng, Y., Yuan, L., Meng, G., Huang, H., Huang, X., Xu, X., Peng, X."c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression". International Journal of Oncology 68.1 (2026): 13.
Chicago
Meng, Y., Yuan, L., Meng, G., Huang, H., Huang, X., Xu, X., Peng, X."c‑Myc‑regulated RPLP0 via the ROS‑mediated JAK2/STAT3 positive feedback loop facilitates hepatocellular carcinoma malignancy progression". International Journal of Oncology 68, no. 1 (2026): 13. https://doi.org/10.3892/ijo.2025.5826
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