Shikonin Induces Ferroptosis through ROS/JNK Pathway to Intervene in the Malignant Behavior of Pancreatic Cancer

Ruifeng QIN; Jiadong XUE; Jia ZHANG; Fan LIU; Shaohui ZHANG; Liyang YIN; Zengjiang YUAN

doi:10.12259/j.issn.2095-610X.S20251005

Volume 46 Issue 10

Oct. 2025

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Article Navigation > Journal of Kunming Medical University > 2025 > 46(10): 44-52

Ruifeng QIN, Jiadong XUE, Jia ZHANG, Fan LIU, Shaohui ZHANG, Liyang YIN, Zengjiang YUAN. Shikonin Induces Ferroptosis through ROS/JNK Pathway to Intervene in the Malignant Behavior of Pancreatic Cancer[J]. Journal of Kunming Medical University, 2025, 46(10): 44-52. doi: 10.12259/j.issn.2095-610X.S20251005

Citation:

Ruifeng QIN, Jiadong XUE, Jia ZHANG, Fan LIU, Shaohui ZHANG, Liyang YIN, Zengjiang YUAN. Shikonin Induces Ferroptosis through ROS/JNK Pathway to Intervene in the Malignant Behavior of Pancreatic Cancer[J]. Journal of Kunming Medical University, 2025, 46(10): 44-52. doi: 10.12259/j.issn.2095-610X.S20251005

Citation:

Ruifeng QIN, Jiadong XUE, Jia ZHANG, Fan LIU, Shaohui ZHANG, Liyang YIN, Zengjiang YUAN. Shikonin Induces Ferroptosis through ROS/JNK Pathway to Intervene in the Malignant Behavior of Pancreatic Cancer[J]. Journal of Kunming Medical University, 2025, 46(10): 44-52. doi: 10.12259/j.issn.2095-610X.S20251005

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Shikonin Induces Ferroptosis through ROS/JNK Pathway to Intervene in the Malignant Behavior of Pancreatic Cancer

doi: 10.12259/j.issn.2095-610X.S20251005

1.
Department of General Surgery III
2.
Operating Room，Handan Central Hospital，Handan Hebei 056001，China

Received Date: 2025-07-18
Available Online: 2025-09-28
Publish Date: 2025-10-28

Abstract

Abstract

Objective To investigate if Shikonin （SKI） can induce ferroptosis via the ROS/JNK pathway to inhibit the malignant behavior of pancreatic cancer. Methods Human pancreatic cancer PANC-1 or BxPC-3 cells were selected. Drug efficacy experiments were established with a blank control group （Con group） and low, medium, and high dose SKI groups （2, 4, 8 μmol/L）. JNK-related mechanism experiments were categorized into a blank control group （Con group）, SKI group, and SKI+JNK inhibitor group （SKI+SP600125 group）. ROS-related mechanism experiments were divided into a blank control group （Con group）, SKI group, and SKI+ROS scavenger group （SKI+NAC group）. Cell viability was assessed using the CCK-8 method to calculate IC₅₀; Transwell experiments evaluated cell migration and invasion capabilities; the C11 BODIPY 581/591 probe was utilized for flow cytometry to detect lipid peroxidation levels, while the FerroOrange fluorescent probe measured ferrous ion levels; ROS levels were determined using a ROS detection kit; the Western blot method identified ferroptosis-related key proteins （SLC7A11, GPX4）, apoptosis-related proteins （Caspase3, PARP）, and JNK pathway proteins （JNK, p-JNK）; an in vivo xenograft tumor model was employed to assess tumor proliferation. Results SKI treatment significantly and dose-dependently inhibited PANC-1 cell viability （IC₅₀: 6.04 μmol/L, P < 0.0001） and BxPC-3 cell viability （IC₅₀: 12.27 μmol/L, P < 0.0001）, and significantly reduced migrating and invasive cell numbers （P < 0.0001）, with migration cell numbers dropping to about 30% of the control group at 8 μmol/L SKI treatment （P < 0.0001）. Mechanistically, SKI induced increased intracellular lipid peroxidation, Fe²⁺ accumulation, and significant ROS production （P < 0.0001）, significantly downregulated SLC7A11 and GPX4 protein expression （GPX4 protein expression reduced to 40% of that in the control group, P < 0.0001）, and activated JNK phosphorylation （p-JNK/JNK ratio increased to 2.8-fold, P < 0.0001）. Pretreatment with the JNK-specific inhibitor SP600125 or ROS scavenger NAC effectively reversed SKI's inhibition of cell viability and downregulation of SLC7A11/GPX4 protein （all P < 0.01）. SKI also inhibited pancreatic cancer tumor cell proliferation in vivo （P < 0.0001）. Conclusion SKI induces ferroptosis by activating the ROS/JNK pathway, thereby inhibiting pancreatic cancer proliferation, migration, and invasion.
- Shikonin,
- Ferroptosis,
- Pancreatic cancer,
- Reactive oxygen species,
- JNK

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