Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: Effect of chemotherapeutic drugs on BLID expression. Treatment with chemotherapeutics increased BLID mRNA expression in (A) MCF-7 and (B) T47D breast cancer cells. Data were normalized to GAPDH, and the fold change in BLID expression was calculated relative to normalized vehicle control. Data are presented as the mean ± SD of two to three replicates with two to three experimental repeats per treatment group. All y-axes denote BLID mRNA expression (fold change vs. vehicle). *P<0.05 (vs. vehicle treatment at the time corresponding to the corresponding experimental treatment time period). DXR, doxorubicin; DTX, docetaxel; 5-FU, 5-fluorouracil; BLID, BH-3 like motif containing inducer of cell death.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: Expressing, Control

Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: Chemotherapeutics induce binding of FOXO proteins to the BLID promoter in MCF-7 cells. Cells were treated with (A) DXR or (B) PTX (+) or a vehicle control (−) as shown, and ChIP-PCR was subsequently performed as shown in the middle panels in (A and B). The left panels in (A and B) show input PCR. The ChIP-PCR assay controls tested were chromatin immunoprecipitated with RNA-Pol II antibody, mouse IgG or rabbit IgG, and using GAPDH or BLID/S1 primers as shown in the right panels in (A and B). (C) ChIP-qPCR. A total of 50 ng of the bound DNA fraction or input DNA was used for qPCR. qPCR data were normalized to input DNA. Data are presented as the mean ± SD of two to three replicates with two to three experimental repeats per treatment group. *P<0.05 (compared with vehicle treatment at the time corresponding to the corresponding experimental treatment time period). DXR, doxorubicin; PTX, paclitaxel; IP, immunoprecipitation; ChIP-PCR, chromatin immunoprecipitation-PCR; qPCR, quantitative PCR; RNA Pol-II, RNA polymerase II; prom., promoter; IgG, immunoglobulin G; BLID, BH-3 like motif containing inducer of cell death.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: Binding Assay, Control, Immunoprecipitation, ChIP-qPCR, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: BLID knockdown in breast cancer cells decreases the sensitivity to chemotherapeutic drugs. (A) Knockdown of BLID expression was confirmed using western blotting. Cells were transfected with the indicated BLID shRNA lentiviral clone or Scr and the cell lysates were analyzed by western blotting using anti-BLID antibody (dilution, 1:10,000). The same blot was also probed with anti-α-Tubulin (dilution, 1:1,000). Data were semi-quantified as shown in the right panel. (B) Knockdown of BLID expression using the sh47 lentiviral clone was further validated using western blotting. The same blot was also probed with anti-GAPDH antibody (dilution, 1:500). (C) Drug dose response. Following BLID knockdown, cells were treated with DXR or vehicle (DMSO) for 24 h and cell viability was assessed using an XTT cell viability assay. The x-axis title of the top graph is identical to that of the bottom graph. Data are presented as the mean ± SD from two to three independent experiments with each data point representing six wells. (D) Left panel: Drug response of MCF-7 cells stably expressing BLID shRNA (sh47) or control scrambled shRNA. Cells were treated with 10 µM 5-FU, 50 nM PTX or 5 nM ETO for 24 h and cell viability was measured. Solid bars, BLID shRNA; empty bars, control scrambled shRNA. Right panel: Reverse transcription-quantitative PCR analysis showing BLID knockdown in BLID shRNA (sh47) vs. scrambled control shRNA-transfected MCF-7 cells. β-Actin served as the housekeeping gene. *P<0.05, **P<0.01, n=3. shRNA/sh, short hairpin RNA; Scr, scrambled control shRNA; Ctl, control; DXR, doxorubicin; 5-FU, 5-fluorouracil; PTX, paclitaxel; ETO, etoposide; BLID, BH-3 like motif containing inducer of cell death.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: Knockdown, Expressing, Western Blot, Transfection, shRNA, Viability Assay, Stable Transfection, Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: RT-qPCR validation of changes in expression of a subset of genes following BLID knockdown. RT-qPCR analysis of several genes in (A) MCF-7 and (B) MDA-MB-231 cells. β-Actin served as the internal control. (A) The y-axis title of the middle graph is identical to the y-axis title of the left graph. (B) The y-axis title of the middle-left graph is identical to the y-axis title of the far-left graph. *P<0.05 and **P<0.01 (BLID shRNA vs. the corresponding Scr shRNA group), n=3. shRNA, short hairpin RNA; Scr, scramble control; Ctl, control; RT-qPCR, reverse transcription-quantitative PCR; BLID, BH-3 like motif containing inducer of cell death.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: Quantitative RT-PCR, Biomarker Discovery, Expressing, Knockdown, Control, shRNA, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: Validation of changes in the expression of a subset of genes in BLID-knockdown breast cancer cells. (A) PI3K-p110α protein expression was increased in BLID shRNA knockdown MCF-7 cells. (B) Left panel: cIAP2/BIRC3 protein expression was increased and DFNA5 expression was decreased in the BLID knockdown MDA-MB-231 cells in the absence or presence of Biri (50 µM; 6 h). Right panel: Reverse transcription-quantitative PCR analysis of BLID expression in MDA-MB-231 cells treated with BLID siRNA or Ctl siRNA. Gene expression was normalized to β-actin as an internal control. **P<0.01 vs. Ctl siRNA, n=3. shRNA, short hairpin RNA; siRNA, small interfering RNA; Ctl, control; Biri, birinapant; BLID, BH-3 like motif containing inducer of cell death; cIAP, cellular inhibitor of apoptosis protein; XIAP, X-linked inhibitor of apoptosis protein.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: Biomarker Discovery, Expressing, Knockdown, shRNA, Reverse Transcription, Real-time Polymerase Chain Reaction, Gene Expression, Control, Small Interfering RNA

Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: BLID is a significant prognostic biomarker in breast and liver cancer as shown by Kaplan-Meier plotter analysis. BLID, BH-3 like motif containing inducer of cell death; RNA-seq, RNA sequencing; FDR, false discovery rate; HR, hazard ratio.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: Biomarker Discovery, RNA Sequencing

Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: Kaplan-Meier analysis of the relationship between BLID expression and overall survival probability in patients with kidney and lung cancer. BLID, BH-3 like motif containing inducer of cell death; RNA-seq, RNA sequencing; FDR, false discovery rate; HR, hazard ratio.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: Expressing, RNA Sequencing

Journal: Oncology Letters

Article Title: BLID is a drug-responsive target of FOXO3a and multi-omics analysis reveals survival mechanisms and therapeutic vulnerabilities in BLID-deficient breast cancer cells

doi: 10.3892/ol.2025.15155

Figure Lengend Snippet: ROC plotter analysis of the predictive response of breast cancer to anthracyclines. ROC P=1.8×10 −5 . ROC, receiver operating characteristic; AUC, area under the curve; TPR, true positive rate; TNR, true negative rate; BLID, BH-3 like motif containing inducer of cell death.

Article Snippet: The BLID shRNA lentiviral plasmids and the scramble control lentiviral plasmid (Addgene cat. no. 1864) were transfected along with the packaging plasmid psPAX2 (Addgene cat. no. 12260) and the envelop expressing plasmid pMD2.G (Addgene cat. no. 12259) into 293T cells using FuGENE HD transfection reagent (Promega Corporations).

Techniques: