ArticleBiochemistry and biophysics reports2026
Comparative effects of a TLR7/8 agonist and doxorubicin on immune checkpoint modulation and apoptosis in 4T1 breast cancer cells.
Article in Biochemistry and biophysics reports, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Background: Breast cancer is still one of the challenges of medical science, despite fast and remarkable advancements in diagnosis and new therapy approaches. This study aims to explore the individual effects of TLR7/8 agonists and to compare their cytotoxic properties with those of the chemotherapy drug doxorubicin on 4T1 breast cancer cells, as well as to evaluate their role in the modulation of immune checkpoint molecules. Methods: 4T1 cancer cells were treated with the TLR7/8 agonist R848 and doxorubicin for a duration of 48 h. First, the viability of the cells was assessed using the MTT method. The relative expression of mRNA for Gal-9, PD-L1, and PVR was analyzed with HPRT serving as a housekeeping gene. Finally, an apoptosis test was conducted to evaluate the cytotoxic effects of R848 and doxorubicin on 4T1 cells. And the wound healing assay was completed in order to assess the migratory potential of 4T1 cells after treatment with R848 and doxorubicin. Results: The expression of the PVR and Gal-9 genes in the group treated with R848 showed a decrease compared to the control group, although this change was not statistically significant. In contrast, the expression of PD-L1 and PVR in the group treated with doxorubicin increased significantly when compared to both the control group and the R848 treated group. Additionally, the total apoptosis rate in both the R848 and doxorubicin treatment groups was significantly higher than that in the control group. After 24 h post-scratch, control 4T1 cells showed about 50 % wound closure, while R848 reduced it to 35 %. Doxorubicin lowered migration to under 10 %. By 48 h, control and R848 nearly closed the wound, unlike the DOX group. Conclusion: R848 and doxorubicin possess anti-proliferative and pro-apoptotic effects on 4T1 cells, doxorubicin effectively induces cell death and boosts immune checkpoint gene expression, while R848 fosters early apoptosis without raising checkpoint ligand expression.
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