Figure 1.
Cell viability values and statistical evaluation of escitalopram oxalate and tamoxifen (positive control) concentrations on MCF-7, MDA-MB-231, and THP-1 cell lines (****P < 0.0001, R2 is coefficient of determination).
The figure consists of six line graphs showing the inhibition percentage for three cell lines—MCF-7, MDA-MB-231, and THP-1—treated with Escitalopram Oxalate or Tamoxifen across various concentrations and repetitions. Each graph contains a curve representing different concentrations (125, 62.5, 31.25, 15.625, 7.8125, and 3.90625) with a control for comparison. Key details are:
1. **Graph a (MCF-7 on Escitalopram Oxalate)**: R² = 0.9994 shows inhibition % over repetitions.
2. **Graph b (MCF-7 on Tamoxifen)**: R² = 0.9977 with similar inhibition % representation.
3. **Graph c (MDA-MB-231 on Escitalopram Oxalate)**: R² = 0.9877 tracks inhibition % over repetitions.
4. **Graph d (MDA-MB-231 on Tamoxifen)**: R² = 0.9967 shows the effect over repetitions.
5. **Graph e (THP-1 on Escitalopram Oxalate)**: R² = 0.9789 with inhibition % data over repetitions.
6. **Graph f (THP-1 on Tamoxifen)**: R² = 0.9899 shows inhibition % change across repetitions.
Each graph shows a significant difference between different concentrations (denoted by **** for all values), indicating statistical significance.
Table 1.
IC50 values of tamoxifen and escitalopram oxalate for MCF-7 and MDA-MB-231 cell lines
This table compares the half maximal inhibitory concentration (IC₅₀) values of tamoxifen and escitalopram oxalate in two different cancer cell lines: MCF-7 and MDA-MB-231.
| Cell line | Tamoxifen (μM) | Escitalopram oxalate (μM) |
|---|---|---|
| MCF-7 | 54.6 | 13.7 |
| MDA-MB-231 | 58.4 | 10.9 |
Figure 2.
MCF-7 and MDA-MB-231 cell lines proliferation curve of 48 h concentrations of tamoxifen and escitalopram oxalate (n = 4, mean ± standard deviation). After 24 h, tamoxifen/escitalopram oxalate was given (arrowed). When the device starts to read again, the cell proliferation according to the concentrations is displayed on the graph hourly.
The image contains two line graphs comparing the effect of Tamoxifen (top graph) and Escitalopram Oxalate (bottom graph) on cell index over time.
**Top Graph (Tamoxifen)**:
- The Y-axis represents the cell index, ranging from -0.5 to 5.5.
- The X-axis shows time in hours, ranging from 0 to 72.
- The graph includes curves for different concentrations of Tamoxifen: 125, 62.5, 31.25, 15.63, 7.91, and 3.91 µM.
- Control and medium conditions are also shown.
- A marked drop in cell index occurs at approximately 24 hours (indicated by a black arrow), followed by a slow recovery for lower concentrations, while higher concentrations show a sustained decline in cell index.
**Bottom Graph (Escitalopram Oxalate)**:
- The Y-axis represents the cell index, ranging from 0 to 3.
- The X-axis shows time in hours, ranging from 0 to 72.
- The graph includes curves for different concentrations of Escitalopram Oxalate: 30, 15, and 7.5 µM.
- Control and medium conditions are also plotted.
- Similar to the top graph, there is a marked drop in cell index around 24 hours (indicated by a black arrow), with subsequent recovery for lower concentrations and continued inhibition for higher doses.
Both graphs illustrate how increasing concentrations of Tamoxifen and Escitalopram Oxalate significantly reduce cell index after a certain point in time.
The image contains two line graphs comparing the effect of Tamoxifen (top graph) and Escitalopram Oxalate (bottom graph) on cell index over time, with control and medium conditions included for comparison.
**Top Graph (Tamoxifen)**:
- The Y-axis represents the cell index, ranging from -0.5 to 2.0.
- The X-axis shows time in hours, ranging from 0 to 72.
- Different concentrations of Tamoxifen are shown: 125, 62.5, 31.25, 15.63, 7.81, and 3.91 µM, as well as control and medium.
- Around 24 hours (marked by a black arrow), a significant drop in cell index occurs for all concentrations, followed by varying degrees of recovery depending on the dose, with higher concentrations leading to continued inhibition and lower concentrations allowing recovery.
**Bottom Graph (Escitalopram Oxalate)**:
- The Y-axis represents the cell index, ranging from 0 to 8.
- The X-axis also represents time in hours, ranging from 0 to 72.
- Concentrations of Escitalopram Oxalate tested are 30, 15, and 7.5 µM, along with control and medium.
- Similar to the top graph, a significant drop in cell index is observed around 24 hours (marked by a black arrow), followed by a varied response in recovery based on the concentration. Higher doses (30 µM) show sustained inhibition, while lower doses exhibit a gradual recovery.
Both graphs emphasize the cytotoxic effects of Tamoxifen and Escitalopram Oxalate over time, particularly after the 24-hour mark, with concentration-dependent inhibition.
Figure 3.
The effect of IC50 values of tamoxifen and escitalopram oxalate on MCF-7 and MDA-MB-231 cell lines by staining with Annexin-V – PI dye in flow cytometry. Q13-LL Live cell, Q13-UL Necrosis, Q13-LR Early Apoptosis, Q13-UR Late Apoptosis.
The image consists of six scatter plots representing flow cytometry results, analyzing apoptosis in MCF-7 and MDA-MB-231 cells treated with Tamoxifen and Escitalopram Oxalate at their IC50 concentrations, compared to controls.
**Top Row (MCF-7 cells)**:
1. **F09 MCF-7 Control**:
- Majority of cells (92.4%) are in the lower-left quadrant (Q13-LL), indicating live cells.
- Minimal apoptosis is seen in the upper-right quadrant (Q13-UR, 2.6%).
2. **F10 MCF-7 Tamoxifen IC50**:
- Apoptosis increases significantly (Q13-UR, 17.9%).
- Live cells decrease to 62.0% (Q13-LL).
3. **F11 MCF-7 Escitalopram Oxalate IC50**:
- Apoptosis in the upper-right quadrant is 3.8%.
- Live cells remain the majority (Q13-LL, 81.0%).
**Bottom Row (MDA-MB-231 cells)**:
1. **F12 MDA Control**:
- Most cells (84.9%) are live (Q13-LL), with minimal apoptosis (Q13-UR, 2.2%).
2. **G01 MDA Tamoxifen IC50**:
- Apoptosis increases to 28.7% in the upper-right quadrant.
- Live cells drop to 62.7% (Q13-LL).
3. **G02 MDA Escitalopram Oxalate IC50**:
- Apoptosis is relatively low (Q13-UR, 1.6%).
- Majority of cells remain live (Q13-LL, 81.4%).
These scatter plots display how Tamoxifen induces significant apoptosis in both MCF-7 and MDA-MB-231 cells, while Escitalopram Oxalate shows comparatively less apoptosis.
Table 2.
Results for IL-1β, IL-6, IL-8, IL-10, and TNF-α in MCF-7 co-culture model (percent positive sign indicates amount secreted)
This table shows the percentage of cytokine secretion (IL-1β, IL-6, IL-8, IL-10, and TNF-α) in THP-1, MCF-7, and co-culture cell lines under control conditions and different treatments (LPS, tamoxifen, escitalopram). It compares how each cell line responds to the treatments in terms of inflammatory cytokine production.
| IL-1β | IL-6 | IL-8 | IL-10 | IL-α | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Cell lines | Substances | % (−) | % (+) | % (−) | % (+) | % (−) | % (+) | % (−) | % (+) | % (−) | % (+) |
| THP-1 | Control | 96.2 | 3.8 | 91.1 | 7.7 | 85.5 | 14.2 | 90.0 | 9.7 | 86.9 | 12.7 |
| 100 ng/mL LPS | 57.6 | 42.4 | 76.4 | 20.1 | 53.7 | 44.4 | 77.3 | 22.1 | 68.6 | 30.9 | |
| MCF-7 | Control | 93.8 | 6.2 | 81.2 | 18.5 | 77.3 | 20.2 | 88.6 | 11.0 | 79.8 | 19.9 |
| Co-culture | Control | 89.6 | 10.4 | 89.4 | 10.3 | 42.7 | 56.1 | 65.8 | 33.3 | 54.8 | 44.2 |
| Tamoxifen IC50 | 94.1 | 5.9 | 80.6 | 19.2 | 80.1 | 18.7 | 87.0 | 12.6 | 78.4 | 21.1 | |
| Escitalopram oxalate IC50 | 97.2 | 2.8 | 88.7 | 11.1 | 83.8 | 15.6 | 88.9 | 10.6 | 87.7 | 12.1 | |
Table 3.
Results for IL-1β, IL-6, IL-8, IL-10, and TNF-α in the MDA-MB-231 co-culture model (percent positive sign indicates amount secreted)
This table presents cytokine secretion results for IL-1β, IL-6, IL-8, IL-10, and TNF-α in THP-1 and MDA-MB-231 cell lines and their co-culture. The table compares control conditions and treatments with 100 ng/mL LPS, tamoxifen, and escitalopram oxalate. It highlights the percentage of cells secreting each cytokine under different conditions, showing variations in immune response across the treatments.
| IL-1β | IL-6 | IL-8 | IL-10 | IL-α | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Cell lines | Substances | % (−) | % (+) | % (−) | % (+) | % (−) | % (+) | % (−) | % (+) | % (−) | % (+) |
| THP-1 | Control | 96.2 | 3.8 | 91.1 | 7.7 | 85.5 | 14.2 | 90.0 | 9.7 | 86.9 | 12.7 |
| 100 ng/mL LPS | 57.6 | 42.4 | 76.4 | 20.1 | 53.7 | 44.4 | 77.3 | 22.1 | 68.6 | 30.9 | |
| MDA-MB-231 | Control | 95.8 | 4.2 | 93.3 | 6.4 | 81.8 | 17.9 | 93.4 | 6.3 | 92.4 | 7.4 |
| Co-culture | Control | 81.9 | 18.1 | 90.3 | 9.4 | 32.0 | 66.7 | 67.5 | 31.5 | 49.3 | 48.6 |
| Tamoxifen IC50 | 95.1 | 4.9 | 85.9 | 13.8 | 83.1 | 16.3 | 89.4 | 10.3 | 75.5 | 24.0 | |
| Escitalopram oxalate IC50 | 97.9 | 2.1 | 83.7 | 15.9 | 82.8 | 16.2 | 90.8 | 8.8 | 87.9 | 11.8 | |
Figure 4.
IL-1β, IL-6, IL-8, IL-10 and TNF-α levels in the MCF-7 and MDA-MB-231 co-culture model. Q16-LR amount of secreted IL-1β, Q20-IL amount of secreted IL-6, Q20-UL amount of secreted TNF-α, Q21-LR amount of secreted IL-8, and Q21-UL amount of secreted IL-10. IL, interleukin.
The image consists of multiple scatter plots (3x4 grid) representing flow cytometry data for IL-1 Beta expression across different cell lines (THP-1, MCF-7, and MDA-MB-231) treated with various conditions including LPS, IL-1 Beta, Tamoxifen, and Escitalopram Oxalate. Each scatter plot is divided into quadrants representing different cell populations, with a specific focus on IL-1 Beta expression.
**Top Row:**
1. **A01 THP-1 IL1B**: A control sample for THP-1 cells treated with IL-1 Beta, showing minimal IL-1 Beta expression in the Q16-UR quadrant (0.0%).
2. **A03 MCF-7 IL1B**: MCF-7 cells treated with IL-1 Beta, also showing negligible IL-1 Beta expression (0.0% in Q16-UR).
3. **A07 THP + MCF7 LPS IL1B Tamox**: Cells treated with LPS, IL-1 Beta, and Tamoxifen, with no detectable IL-1 Beta expression (Q16-UR, 0.0%).
**Middle Row:**
1. **F03 THP-1 100ng LPS IL1B**: THP-1 cells treated with 100ng LPS and IL-1 Beta, with no observable IL-1 Beta expression (Q16-UR, 0.0%).
2. **G10 MCF + THP LPS IL1B**: Mixed MCF and THP-1 cells treated with LPS and IL-1 Beta, showing no detectable expression of IL-1 Beta (Q16-UR, 0.0%).
3. **A08 THP + MCF7 LPS IL1B Escit**: Similar combination as A07 but with Escitalopram, again with no IL-1 Beta expression in Q16-UR (0.0%).
**Bottom Row:**
1. **A01 THP-1 IL1B (Replicate)**: Similar to the first plot, showing 1.2% IL-1 Beta expression in the lower right quadrant (Q16-LR).
2. **A04 MDA231 K IL1B**: MDA-MB-231 cells treated with IL-1 Beta, also showing negligible IL-1 Beta expression.
3. **A09 THP + MDA231 LPS IL1B Tamox**: Co-treatment of THP-1 and MDA-MB-231 cells with LPS, IL-1 Beta, and Tamoxifen, showing no IL-1 Beta expression in Q16-UR (0.0%).
Each plot highlights a focus on the lower quadrants (Q16-LL and Q16-LR), representing live cells with minimal IL-1 Beta expression in most treatment conditions.
The image contains a grid of 6x5 flow cytometry scatter plots showing the expression of IL-6 and IL-10 across different cell types (THP-1, MCF-7, and MDA-MB-231), treated with LPS, IL-6, IL-10, Tamoxifen, and Escitalopram Oxalate. The plots are divided by quadrants, representing different populations of cells and cytokine expression levels.
**Top Three Rows (IL-6 Expression)**:
1. **A11 THP-1 IL6** to **D06 THP-1 IL6**:
- These plots show IL-6 expression in THP-1 and MCF-7 cells treated with various conditions, including LPS, IL-6, Tamoxifen, and Escitalopram Oxalate.
- Cells treated with IL-6 (e.g., A11, A12, and A17) show minimal IL-6 expression in the upper-right quadrant (Q21-UR).
- Co-treatments with LPS and Tamoxifen, or LPS and Escitalopram, show slight variations in IL-6 expression but remain mostly clustered in the lower quadrants, indicating low IL-6 expression.
**Bottom Three Rows (IL-10 Expression)**:
1. **C07 THP-1 IL10** to **D04 THP-1 IL10**:
- These plots focus on IL-10 expression under similar treatments (LPS, IL-10, Tamoxifen, and Escitalopram Oxalate).
- The upper-right quadrants (Q21-UR) represent the fraction of cells expressing IL-10, which remains low across most treatments, with slight increases in certain conditions (e.g., C08, C12).
- Co-treatments with Tamoxifen and Escitalopram again show minimal effect on IL-10 expression in both cell types.
Overall, the scatter plots indicate low levels of IL-6 and IL-10 expression under these experimental conditions, with little variation in the upper-right quadrants (Q21-UR) across the different treatments.