Journal: ACS Omega

Article Title: APG-115 Induces SLC7A11-Mediated Ferroptosis and Upregulates PD-L1 Expression in Thyroid Cancer

doi: 10.1021/acsomega.5c04710

Figure Lengend Snippet: APG-115 exhibits therapeutic effects on TC both in vitro and in vivo. (A) CCK-8 assay demonstrating the relative cell viability of TPC-1 and B-CPAP cells at different concentrations of APG-115. (B) Cell scratch assay of B-CPAP and TPC-1 cells treated with APG-115 at 0 and 24 h. Scar bar = 100 μM. (C) Photograph of the tumors from the control and APG-115 treatment groups. (D,E) Tumor volume and weight of the mice from the control and APG-115 treatment groups. * p < 0.05 versus control group. Data are expressed as mean ± SEM, n = 5.

Article Snippet: We measured the cell viability of TC cell lines B-CPAP and TPC-1 following APG-115 treatment using a CCK-8 kit (HY-K0301, MCE).

Techniques: In Vitro, In Vivo, CCK-8 Assay, Wound Healing Assay, Control

Journal: ACS Omega

Article Title: APG-115 Induces SLC7A11-Mediated Ferroptosis and Upregulates PD-L1 Expression in Thyroid Cancer

doi: 10.1021/acsomega.5c04710

Figure Lengend Snippet: APG-115 inhibits the growth of TC patient-derived organoids. (A) Images of TC patient-derived organoids after APG-115 treatment. Scale bar = 100 μM. (B) Concentration–response viability curves of organoids following APG-115 treatment for 5 days. (C) IC 50 values of TC-derived organoids in APG-115 treatment.

Article Snippet: We measured the cell viability of TC cell lines B-CPAP and TPC-1 following APG-115 treatment using a CCK-8 kit (HY-K0301, MCE).

Techniques: Derivative Assay, Concentration Assay

Journal: ACS Omega

Article Title: APG-115 Induces SLC7A11-Mediated Ferroptosis and Upregulates PD-L1 Expression in Thyroid Cancer

doi: 10.1021/acsomega.5c04710

Figure Lengend Snippet: APG-115 treatment upregulates the expression levels of PD-L1, p53, and MDM2 in TC cells. (A) WB assay detected the proteins of PD-L1, p53, and MDM2 in TPC-1 and B-CPAP cells after treatment with APG-115 at 0.125, 0.25, 0.5, 1.0, and 2.0 μM. (B)­Relative expression levels of MDM2, PD-L1, and p53 proteins in BCPAP and TPC-1 cells. ns = nonsignificant, * p < 0.05, ** p < 0.01, *** p < 0.001 versus control group. Data are expressed as mean ± SEM, n = 3. (C) Immunohistochemical staining of the PD-L1 and MDM2 from the tissues of control and APG-115-treated groups. Scale bar = 20 μM. (D) Percentage of immunohistochemically positive area of MDM2 and PD-L1 in mouse tissues from control and APG-115-treated groups. * p < 0.05, *** p < 0.001 versus control group. Data are expressed as mean ± SEM, n = 3. (E) Flow cytometry analysis demonstrating the PD-L1 expression levels in TPC-1 cells from control and APG-115 treatment groups. (F) Correlation analysis of PD-L1 expression level and the p53 pathway in TC.

Article Snippet: We measured the cell viability of TC cell lines B-CPAP and TPC-1 following APG-115 treatment using a CCK-8 kit (HY-K0301, MCE).

Techniques: Expressing, Control, Immunohistochemical staining, Staining, Flow Cytometry

Journal: ACS Omega

Article Title: APG-115 Induces SLC7A11-Mediated Ferroptosis and Upregulates PD-L1 Expression in Thyroid Cancer

doi: 10.1021/acsomega.5c04710

Figure Lengend Snippet: (A) Images of TPC-1 and B-CPAP cells after APG-115 treatment at different concentrations detected by the C11-BODIPY581/591 probe. Scale bar = 100 μM. (B,C) WB assay demonstrated the proteins of SLC7A11 and GPX4 levels following APG-115 treatment in TPC-1 and B-CPAP cells; Relative expression levels of SLC7A11 and GPX4 proteins in BCPAP and TPC-1 cells following APG-115 treatment. ns = nonsignificant, * p < 0.05, ** p < 0.01, *** p < 0.001 versus control group. Data are expressed as mean ± SEM, n = 3. (D) Images of TPC-1 and B-CPAP cells in control, APG-115, FER-1, and combined treatment groups were detected by the C11-BODIPY581/591 probe. Scale bar = 100 μM. (E) WB assay and the relative expression levels of SLC7A11 and PD-L1 proteins in control, APG-115, FER-1, and combined treatment groups, both in TPC-1 and B-CPAP cells. ns = nonsignificant, * p < 0.05, ** p < 0.01, *** p < 0.001. Data are expressed as mean ± SEM, n = 3.

Article Snippet: We measured the cell viability of TC cell lines B-CPAP and TPC-1 following APG-115 treatment using a CCK-8 kit (HY-K0301, MCE).

Techniques: Expressing, Control

Journal: Anti-Cancer Drugs

Article Title: APG-115 synergizes with bortezomib to induce apoptosis in cervical cancer cells

doi: 10.1097/CAD.0000000000001735

Figure Lengend Snippet: APG-115 activates p53 expression and function. (a) HeLa cells and (b) SiHa cells were treated with different concentrations of APG-115 for 12 h, the relative mRNA expression levels of MDM2 , TP53 , and CDKN1A(p21 ) were measured by RT-qPCR. Experiments were performed in triplicate and repeated at least three times. (c) HeLa cells were treated with 5 μM APG-115 for different time intervals, and the protein expression levels of MDM2, p53, and p21 were analyzed by WB. β-actin served as the internal reference protein. The data shown are representative images from two independent experiments. (d) The quantification analysis of WB bands in (c) was performed using ImageJ software. WB, western blotting

Article Snippet: APG-115 was provided by Ascentage Pharma Co., Ltd. (Suzhou, China), and bortezomib was purchased from MedChemExpress (MCE).

Techniques: Expressing, Quantitative RT-PCR, Software, Western Blot

Journal: Anti-Cancer Drugs

Article Title: APG-115 synergizes with bortezomib to induce apoptosis in cervical cancer cells

doi: 10.1097/CAD.0000000000001735

Figure Lengend Snippet: APG-115 induces apoptosis in cervical cancer cells. (a) The inhibitory effect of APG-115 on the proliferation of HeLa cells and (b) SiHa cells was measured using the MTS assay. Apoptosis induction in (c) SiHa cells and (d and e) HeLa cells was assessed by FACS analysis. The graphs display the mean percentages of Annexin-V/PI-positive cells, with error bars representing the SD. Experiments were conducted in duplicate and repeated at least three times. (f) Representative Western blot images showing the expression levels of AKT, p-AKT, ERK, and p-ERK; (g) BCL-2, BCL-xL, and MCL-1; and (h) BAK, BAX, and BIM proteins in lysates of HeLa cells treated with different concentrations of APG-115. β-actin was used as a loading control. (i–k) Quantification of the protein bands in (f–h) was performed using ImageJ software. The band intensities were normalized to the internal control β-actin. Statistical analysis was performed using GraphPad Prism 8.0 software and Student’s t -test. A P -value of <0.05 was considered statistically significant (* P < 0.05; ** P < 0.01; and *** P < 0.001).

Article Snippet: APG-115 was provided by Ascentage Pharma Co., Ltd. (Suzhou, China), and bortezomib was purchased from MedChemExpress (MCE).

Techniques: MTS Assay, Western Blot, Expressing, Control, Software

Journal: Anti-Cancer Drugs

Article Title: APG-115 synergizes with bortezomib to induce apoptosis in cervical cancer cells

doi: 10.1097/CAD.0000000000001735

Figure Lengend Snippet: APG-115 synergizes with bortezomib to induce apoptosis in cervical cancer cells. (a) The percentage of apoptotic cells measured by FACS analysis in HeLa cells treated with APG-115 and bortezomib for 48 h. Experiments were performed in duplicate and repeated at least three times. Data are presented as means ± SD. (b) CI values were calculated using Compusyn software to evaluate the combination effect in panel (a). CI values indicate the degree of synergy: green represents synergy or strong synergy, and red indicates slight synergy. (c) Representative western blots showing the expression of MDM2, p53, p21, and BCL-2 proteins in HeLa cells treated with APG-115 and bortezomib for 6 h. β-actin was used as a loading control. (d) Quantification of the Western blot bands in panel (c) was performed using ImageJ software. Significance of the different treatments was assessed using GraphPad Prism 8.0, the two-way ANOVA analysis. A P -value of <0.05 was considered statistically significant, * P < 0.05; ** P < 0.01; *** P < 0.001; and **** P < 0.0001.

Article Snippet: APG-115 was provided by Ascentage Pharma Co., Ltd. (Suzhou, China), and bortezomib was purchased from MedChemExpress (MCE).

Techniques: Software, Western Blot, Expressing, Control

Journal: Anti-Cancer Drugs

Article Title: APG-115 synergizes with bortezomib to induce apoptosis in cervical cancer cells

doi: 10.1097/CAD.0000000000001735

Figure Lengend Snippet: Evaluation of the antitumor effect of APG-115 in mouse model. (a) Cervical cancer tumor-bearing nude mice were treated with APG-115, bortezomib, or a combination of APG-115 and bortezomib, with PBS used as a control. Tumor volumes were measured continuously over 40 days. (b) Tumor sizes in the different experimental groups. (c) Statistical analysis of tumor weights in the different experimental groups. (d) Pathological changes in the liver and kidney tissues of mice from the different experimental groups. Representative images of (e) Ki-67, (f) TUNEL, (g) p21and BCL-2 staining in tumor tissues from the four groups (magnification: 400×). (h–k) Quantification of the percentage of (e) Ki-67-positive, (f) TUNEL-positive, (g) p21-positive and BCL-2-positive cells in tumor tissues. Statistical analysis was performed using an unpaired Student’s t -test with Welch’s correction. Symbols denote statistical significance: *compared to the control group; #compared to the bortezomib group; &compared to the APG-115 group. *, #, & P < 0.05; **, ##, && P < 0.01; and ***, ###, &&& P < 0.001. Nonsignificant results are denoted by ‘ns’.

Article Snippet: APG-115 was provided by Ascentage Pharma Co., Ltd. (Suzhou, China), and bortezomib was purchased from MedChemExpress (MCE).

Techniques: Control, TUNEL Assay, Staining

Journal: Anti-Cancer Drugs

Article Title: APG-115 synergizes with bortezomib to induce apoptosis in cervical cancer cells

doi: 10.1097/CAD.0000000000001735

Figure Lengend Snippet: The proposed mechanism of APG-115 and bortezomib synergistically inducing apoptosis in cervical cancer cells. The schematic illustrates multiple potential mechanisms through which APG-115 and bortezomib collaboratively contribute to the synergistic induction of cervical cancer cell apoptosis.

Article Snippet: APG-115 was provided by Ascentage Pharma Co., Ltd. (Suzhou, China), and bortezomib was purchased from MedChemExpress (MCE).

Techniques: