Background: Cancer remains a major global health challenge, with treatment efficacy often limited by drug resistance and adverse effects. Drug repurposing offers promising opportunities for developing novel anticancer strategies. This study evaluated the cytotoxic, antiproliferative, and pro-apoptotic effects of metformin and caffeine, administered individually and in combination, in human cancer cell lines, as well as their potential interaction mechanisms. Methods: Human cervical carcinoma (HeLa), lung adenocarcinoma (A549), and colorectal carcinoma (HT29) cell lines were treated with metformin (0.05–50 mM) and caffeine (0.5–5 mM), either alone or in combination, for 24 and 48 h. Cell viability and proliferation were assessed using Trypan Blue and sulforhodamine B (SRB) assays. Apoptosis was analyzed by Annexin V/propidium iodide flow cytometry, and p53 expression in HeLa cells was determined by ELISA. Statistical analysis was performed using a one-way ANOVA followed by Tukey’s post hoc test. Results: Metformin induced dose- and time-dependent cytotoxicity in all tested cell lines, with the lowest IC₅₀ values observed in HeLa and A549 cells after 48 h (2.28 and 3.30 mM, respectively; p < 0.05). Caffeine showed moderate antiproliferative activity, with the strongest effects observed at 2.03 mM in HeLa cells and 2.01 mM in HT29 cells (p < 0.05). The combined treatment produced effects that varied depending on both the cell line and exposure time. At earlier time points, transient synergistic effects were observed in certain cell lines, particularly HeLa cells; however, these effects were not sustained over time. With prolonged exposure, the interaction shifted predominantly toward antagonistic effects, indicating the reduced overall efficacy of the combination compared with the expected additive outcomes. Increased apoptosis and elevated p53 expression further supported the activation of tumor-suppressive pathways. Conclusions: Metformin exhibited significant anticancer activity in vitro, supporting its potential repurposing in oncology. However, the addition of caffeine did not uniformly enhance its efficacy and appeared to exert context-dependent effects. Further in vivo studies are required to confirm the clinical relevance of these findings.

Background:Cancer remains a major global health challenge, with treatment efficacy li-mited by drug resistance and adverse effects. Drug repurposing offers opportunities for novel anticancer strategies. This study evaluated the cytotoxic, antiproliferative, and pro-apoptotic effects of metformin and caffeine, alone and in combination, in human cancer cell lines, and their potentialinteraction mechanisms. Methods:Human cervical carcinoma (HeLa), lung adenocarcinoma (A549), and colorectal carcinoma (HT29) cell lines were treated with metformin (0.05–50 mM) and caffeine (0.5–5 mM), alone or combined, for 24 and 48 h. Cell viability and proliferation were assessed using Trypan Blue and sulforhodamine B (SRB) assays. Apoptosis was analyzed by Annexin V/propidium iodide flow cytometry, and p53 expression in HeLa cells was determined by ELISA. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. Results:Metformin induced dose- and time-dependent cytotoxicity in all tested cell lines, with the lowest IC₅₀ values observed in HeLa and A549 cells after 48 h (2.28 and 3.30 mM, respectively; p < 0.05). Caffeine showed moderate antiproliferative activity, with the strongest effects at 2.03 mM in HeLa and 2.01 mM in HT29 cells (p < 0.05). The combined treatment produced effects that varied depending on both the cell line and exposure time. At earlier time points, transient synergistic effects were observed in certain cell lines, particularly HeLa; however, these effects were not sustained over time. With prolonged exposure, the interaction shifted toward predominantly antagonistic effects, indicating a reduced overall efficacy of the combination compared to expected additive outcomes.Increased apoptosis and elevated p53 expression further support the activation of tumor-suppressive pathways. Conclusions:Metformin exhibits significant anticancer activity in vitro, supporting met-formin repurposing in oncology. However,the addition of caffeine does not uniformly enhance its efficacy and appears to exert context-dependent effects.Further in vivo studies are required to confirm its clinical relevance. Keywords:AMPK;Antitumor activity; Apoptosis; Caffeine; Cancer cell lines;Chou–Talalay method; Drug repurposing;Docking;Metfomin;Molecular docking; p53;

Background/Objectives: Lumbar radiculopathy can be managed non-surgically or surgically; however, comparative changes in quality of life and functional status between these treatment pathways remain clinically relevant. Therefore, this study aimed to compare six-month outcomes in patients treated non-surgically and those treated with lumbar microdiscectomy followed by rehabilitation. Methods: A prospective, non-randomized comparative cohort study included 100 patients with discogenic lumbar radiculopathy: 50 treated non-surgically and 50 treated by lumbar microdiscectomy. Both groups underwent medical rehabilitation and were assessed at admission, three months, and six months using the Short Form-36 Health Survey (SF-36), including the Physical Component Summary (PCS) and Mental Component Summary (MCS), and the Oswestry Disability Index (ODI). Results: PCS improved from 35.5 to 50.8 in the non-surgical group and from 28.8 to 49.2 in the surgical group. PCS was significantly higher in non-surgically treated patients at admission (p < 0.001) and three months (p < 0.05), whereas the difference was not significant at six months (p = 0.064). No between-group differences were found for MCS at admission, three months, or six months (p = 0.104, p = 0.871, and p = 0.323, respectively). ODI decreased from 51.5% to 22.5% in the non-surgical group and from 56.1% to 23.7% in the surgical group, without significant between-group differences at any assessment (p = 0.109, p = 0.162, and p = 0.427). Conclusions: Both treatment pathways were associated with substantial improvement. The initial between-group difference in physical quality of life diminished during follow-up, while mental quality of life and disability outcomes were similar between groups.

Background: Cancer remains a major global health challenge, with treatment efficacy limited by drug resistance and adverse effects. Drug repurposing offers opportunities for novel anticancer strategies. This study evaluated the cytotoxic, antiproliferative, and pro-apoptotic effects of metformin and caffeine, alone and in combination, in human cancer cell lines, and their potentialinteraction mechanisms. Methods: Human cervical carcinoma (HeLa), lung adenocarcinoma (A549), and colorectal carcinoma (HT29) cell lines were treated with metformin (0.05–50 mM) and caffeine (0.5–5 mM), alone or combined, for 24 and 48 h. Cell viability and proliferation were assessed using Trypan Blue and sulforhodamine B (SRB) assays. Apoptosis was analyzed by Annexin V/propidium iodide flow cytometry, and p53 expression in HeLa cells was determined by ELISA. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test. Results: Metformin induced dose- and time-dependent cytotoxicity in all cell lines, with the lowest IC₅₀ values in HeLa and A549 cells after 48 h (2.28 and 3.30 mM; p < 0.05). Caffeine showed moderate antiproliferative activity, with strongest effects at 2.03 mM in HeLa and 2.01 mM in HT29 cells (p < 0.05). Combined treatment demonstrated variable effects depending on the cell line and treatment duration, with limited synergistic interaction observed only under specific conditions, while predominantly antagonistic effects were detected overall. Increased apoptosis and elevated p53 expression suggest activation of tumor-suppressive pathways. Conclusions: Metformin exhibits significant anticancer activity in vitro, supporting metformin repurposing in oncology. However,the addition of caffeine does not uniformly enhance its efficacy and appears to exert context-dependent effects.Further in vivo studies are required to confirm its clinical relevance.

Background: Metabolic reprogramming enables cancer cells to adapt to energetic stress and sustain proliferation. Simultaneous inhibition of glycolysis and mitochondrial respiration has emerged as a promising strategy to overcome metabolic plasticity. This study evaluated the antiproliferative effects of the glycolytic inhibitor 2-deoxy-D-glucose (2-DG) alone and in combination with metformin in human cancer cell lines. Methods: Human cervical carcinoma (HeLa), lung adenocarcinoma (A549), colorectal adenocarcinoma (HT-29), and normal lung fibroblasts (MRC-5) were treated with 2-DG or metformin individually, or with metformin in the presence of a fixed concentration of 2-DG (1 mM). Cell viability was assessed using the sulforhodamine B assay after 24 and 48 h. IC₅₀ values were calculated by nonlinear regression, Dose Reduction Index (DRI) analysis evaluated sensitization to metformin, and molecular docking was performed to investigate interactions with selected metabolic targets. Results: Both 2-DG and metformin inhibited cell proliferation in a concentration- and time-dependent manner. HeLa cells were the most sensitive to glycolytic inhibition, while A549 and HT-29 cells showed moderate susceptibility. Co-treatment with 2-DG significantly enhanced metformin activity, reducing its IC₅₀ in HeLa cells from 6.04 to 2.00 mM after 24 h and from 2.28 to 1.56 mM after 48 h. DRI analysis demonstrated increased sensitivity to metformin in all cancer cell lines, particularly HT-29 and A549, whereas normal MRC-5 fibroblasts remained comparatively less affected. Molecular docking revealed favorable binding of both 2-DG and metformin to proteins involved in cellular energy metabolism. Conclusions: Combined inhibition of glycolysis and mitochondrial respiration potentiates the antiproliferative effects of metformin, increases metabolic vulnerability in cancer cells, and is supported by molecular docking evidence of interactions with metabolic targets, while showing limited toxicity toward normal fibroblasts. These findings support dual metabolic targeting as a promising therapeutic strategy and warrant further mechanistic and in vivo studies.