Postbiotics Combination Synergises the Antiproliferative Effects of Doxorubicin in Gastric Cancer Cells: A Cellular and Molecular Deep Dive

Figures

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  Figure 1. CI values of different inhibitory concentrations of APB+Dox on AGS gastric cancer cells.

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  Figure 2. Pathway enrichment analysis using differentially expressed proteins in Dox-treated AGS cells compared to control cells. (A) Volcano plot showing significantly regulated proteins (absolute log2FC ≥ 0.58 and Q ≤ 0.05). (B) Graphical summary of top predictions in ingenuity pathway analysis (IPA) interpret report as derived from IPA core analysis.

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  Figure 3. A schematic representation of the major pathway of rRNA processing in the nucleolus and cytosol. Proteins indicated in red were significantly upregulated, while proteins indicated in blue were downregulated in Dox-treated AGS cells compared to control untreated cells. The figure was generated by Qiagen Ingenuity Pathway Analysis (IPA).

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  Figure 4. Pathway enrichment analysis using differentially expressed proteins in APB+Dox-treated AGS cells compared to APB and Dox (Q ≤ 0.05). (A) Volcano plot showing significantly regulated proteins (absolute log2FC ≥ 0.58 and Q ≤ 0.05). (B) Graphical summary of top predictions in an IPA Interpret report presented in the form of a simple network.

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  Figure 5. Network analysis of differentially expressed proteins involved in tumour progression following APB+Dox treatment in AGS cells compared to monotreatments APB and Dox.

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  Figure 6. Flow cytometric assessment of the apoptotic profiles of the AGS gastric cancer cells after 24 h of treatment. (A) The live, early apoptotic, late apoptotic, and necrotic cell percentages after 24 h treatment with APB (3000 µg/mL), Dox (0.54 µg/mL), APB+Dox (3000.27 µg/mL), and control (n = 4). ** Indicates 0.01 < p-value < 0.05; *** indicates p < 0.001; **** indicates p < 0.0001 compared to the negative control. (B) Represented are the density plots of each drug treatment that is most representative of the average data from the flow cytometric analyses, with Q3-1 = necrotic cells, Q3-2 = late-stage apoptotic cells, Q3-3 = live cells, and Q3-4 = early-stage apoptotic cells.

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  Figure 7. Fold change in ROS generation following treatment with various concentrations: 3000:0.27 μg/mL, 1500:0.14 μg/mL, and 750:0.07 μg/mL of APB+Dox. Additionally, Doxorubicin (0.54, 0.27, 0.14 μg/mL) and tert-Butyl hydroperoxide (TBHP) (22.5 μg/mL or 250 μM) are included for comparative purposes. The values are expressed as mean ± SD. **** indicates p < 0.0001 compared to untreated control.