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Archive > Volume 33 Issue 2 > 2026,33(2):181-189. DOI:10.3872/j.issn.1007-385X.2026.02.010 Prev Next
Basic Research
Mitochondrial DNA (mtDNA) partial depletion induces malignant transformation and immune escape in colonic epithelial cell NCM460
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Abstract:
[Abstract] Objective: To clarify the effects of mitochondrial DNA (mtDNA) partial depletion on colonic epithelial cell NCM460. Methods: Mitochondrial DNA partially depleted cell model (mtDNA-reduced) and reversion model (reverted) were established by ethidium bromide (EB) induction. Mitochondrial function was evaluated by CCK-8 (Cell Counting Kit-8) assay, reactive oxygen species (ROS) and mitochondrial membrane potential (JC-1) detection. Metabolic phenotypes were assessed by glucose, lactate and glycolytic key enzyme activity assays. Cell invasion and epithelial-mesenchymal transition (EMT) were detected by Transwell invasion assay and Western blot. Anoikis resistance was examined by polyHEMA coated culture. The PI3K/Akt/mTOR signaling pathway and programmed death ligand 1 (PD-L1) expression were detected, and verified by PI3K inhibitor LY294002. Results: GEPIA (Gene Expression Profiling Interactive Analysis) database analysis showed that mtDNA-encoded genes were down-regulated in colorectal cancer (P < 0.05). Tissue microarray showed that cytochrome coxidase subunit 1 (COX1) protein was highly expressed in normal colonic tissues but lowly expressed in cancer tissues (P < 0.05). Mitochondrial DNA-reduced NCM460 cells showed impaired mitochondrial function (decreased growth rate, reduced ROS generation, and decreased mitochondrial membrane potential, P < 0.05), exhibited the Warburg effect (increased glucose uptake, enhanced lactate secretion, and increased glycolytic key enzyme activity, P < 0.05), enhanced invasion capacity (P < 0.05) with EMT, and resisted anoikis (P < 0.05). The PI3K/Akt/mTOR pathway was activated and PD-L1 expression was upregulated in these cells. After treatment with LY294002, the glucose content showed a decreasing trend (P > 0.05), the cell activity decreased (P < 0.05), and PD-L1 expression was downregulated (P < 0.05). Conclusion: Partial depletion of mtDNA in colonic epithelial cell NCM460 can induce malignant transformation and immune escape.
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