Chinese Journal of Tissue Engineering Research ›› 2026, Vol. 30 ›› Issue (36): 9572-9579.doi: 10.12307/2026.920

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Inducing cuproptosis in pancreatic β cells: regulating homocysteine via targeting SLC39A14

Li Shujuan1, Ma Lingju2, 3, Li Xinru4, Sun Rui4, Ma Ben4, Qi Ning4, Zhang Qifan4, Liu Li4, Chai Yuee5, Ma Shengchao3, 4   

  1. 1Emergency Department, 2Department of Geriatrics and Special Needs Medicine, General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 3Key Laboratory of Metabolic Cardiovascular Disease Research of National Health and Wellness Committee, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 4Inspection College of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China; 5College of Pharmacy, Guizhou Medical University, Guiyang 550000, Guizhou Province, China
  • Received:2025-11-06 Revised:2026-03-25 Online:2026-12-28 Published:2026-05-25
  • Contact: Ma Shengchao, PhD, Associate professor, Key Laboratory of Metabolic Cardiovascular Disease Research of National Health and Wellness Committee, Yinchuan 750004, Ningxia Hui Autonomous Region, China; Inspection College of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China Co-corresponding author: Chai Yuee, PhD, Associate professor, College of Pharmacy, Guizhou Medical University, Guiyang 550000, Guizhou Province, China
  • About author:Li Shujuan, MS, Physician, Emergency Department of General Hospital of Ningxia Medical University, Yinchuan 750004, Ningxia Hui Autonomous Region, China
  • Supported by:
    National Natural Science Foundation of China (Young Scientist Program), No. 81900273 (to MSC); National Natural Science Foundation of China (General Program), Nos. 82060139 and 82270492 (both to MSC); Ningxia Natural Science Foundation (Excellent Young Program), No. 2023AAC05035 (to MSC); Key Project of Ningxia Medical University, No. XZ2022004; Ningxia Medical University College Student Innovation and Entrepreneurship Training Program, No. S202210752015 (to TR [principal investigator] and MSC [advisor])

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Abstract: BACKGROUND: Hyperhomocysteinemia is closely associated with pancreatic β-cell dysfunction and insulin resistance, but its specific molecular mechanism remains to be elucidated.
OBJECTIVE: To investigate the potential mechanism by which piR-000699 targets SLC39A14 to regulate homocysteine-induced cuproptosis in pancreatic β cells.
METHODS: The mouse pancreatic β-cell line Min6 was cultured in vitro and divided into a control group (0 μmol/L homocysteine) and homocysteine groups (treated with 40, 80, 100, and 120 μmol/L homocysteine, respectively). A concentration of 100 μmol/L homocysteine was selected as the optimal dose for intervening in pancreatic β cells. qRT-PCR was used to detect the baseline expression of piR-000699 in homocysteine-treated cells. Western blot was used to detect the expression of cuproptosis-related proteins FDX1 and HSP70, as well as the piR-000699 downstream target gene SLC39A14, in homocysteine-induced pancreatic β cells. A copper ion assay kit was used to measure intracellular Cu²⁺ levels in pancreatic β cells. Cell transfection groups were as follows: (1) control group, homocysteine group (100 μmol/L), homocysteine+mimics-NC group, homocysteine+piR-000699 mimics group; (2) control group, homocysteine group (100 μmol/L homocysteine), homocysteine+inhibitor-NC group, homocysteine+piR-000699 inhibitor group. qRT-PCR was used to detect piR-000699 mRNA expression after transfection with piR-000699 inhibitor and piR-000699 mimics. Western blot was used to detect the protein expression of FDX1, HSP70, and SLC39A14 after transfection with piR-000699 inhibitor and piR-000699 mimics. A copper ion assay kit was used to measure Cu²⁺ expression levels after transfection. Wild-type and mutant dual-luciferase gene vectors for SLC39A14 were constructed to verify the targeted binding relationship between piR-000699 and SLC39A14.
RESULTS AND CONCLUSION: (1) The optimal concentration of homocysteine for inducing pancreatic β-cell injury was 100 μmol/L. (2) Compared with the control group, piR-000699 mRNA expression was increased in the homocysteine group (P < 0.01), and the expression of cuproptosis-related proteins FDX1 and HSP70, as well as Cu²⁺ levels, were all increased (P < 0.001). (3) After interfering with piR-000699 in pancreatic β cells, homocysteine further downregulated the protein expression of FDX1, HSP70, and SLC39A14 (P < 0.01), while intracellular Cu²⁺ expression levels were decreased (P < 0.01). (4) After overexpressing piR-000699 in pancreatic β cells, homocysteine further upregulated the protein expression of FDX1, HSP70, and SLC39A14 in the cells (P < 0.01), while intracellular Cu²⁺ levels were significantly increased (P < 0.001). (5) Dual-luciferase reporter assay results indicated that, compared with mimics-NC, the relative luciferase activity was significantly increased in pancreatic β cells co-transfected with the SLC39A14 wild-type plasmid and piR-000699 mimics (P < 0.01). In conclusion, these findings indicate that piR-000699 can modulate the expression of cuproptosis-related proteins by targeting the SLC39A14 gene during homocysteine-induced cuproptosis in pancreatic β cells, providing new targets and strategies for the treatment and prevention of diabetes.

Key words: piRNA, SLC39A14, pancreatic β cells, homocysteine, cuproptosis

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