Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly reduces melanoma cell viability and proliferation. STAT1 mRNA expression was analyzed in (A) A375 and (B) RPMI-7951 melanoma cells post-STAT1 knockdown using siRNA. (C) Western blotting was used to assess STAT1 protein levels in melanoma cell lines following siRNA-mediated knockdown. Cell viability was evaluated after STAT1 knockdown in (D) A375 and (E) RPMI-7951 cells using Cell Counting Kit-8 assays. (F) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown. (G) Quantification of colony formation assay results. **P<0.01 vs. si-NC group. siRNA, small interfering RNA; STAT1, signal transducer and activator of transcription 1; si-STAT1, siRNA targeting STAT1; si-NC, negative control siRNA.

Article Snippet: The melanoma cell lines A2058 (cat. no. CRL-3601), SK-MEL-1 (cat. no. HTB-67), A375 (cat. no. CRL-1619) and RPMI-7951 (cat. no. HTB-66), as well as normal human epidermal melanocytes (HEM; cat. no. PCS-200-013) and 293T cells (cat. no. CRL-3216) were obtained from American Type Culture Collection.

Techniques: Knockdown, Expressing, Western Blot, Cell Counting, Colony Assay, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 knockdown significantly promotes melanoma cell apoptosis and inhibits migration. (A and B) Apoptosis was assessed and quantified in A375 and RPMI-7951 cells following STAT1 knockdown using flow cytometry. (C) Transwell migration assays were conducted to evaluate cell migration following STAT1 knockdown. Magnification, ×200. (D) Quantification of migration capacity in A375 and RPMI-7951 cells following STAT1 knockdown. *P<0.05 vs. si-NC. STAT1, signal transducer and activator of transcription 1; si-STAT1, small interfering RNA targeting STAT1; si-NC, negative control small interfering RNA.

Article Snippet: The melanoma cell lines A2058 (cat. no. CRL-3601), SK-MEL-1 (cat. no. HTB-67), A375 (cat. no. CRL-1619) and RPMI-7951 (cat. no. HTB-66), as well as normal human epidermal melanocytes (HEM; cat. no. PCS-200-013) and 293T cells (cat. no. CRL-3216) were obtained from American Type Culture Collection.

Techniques: Knockdown, Migration, Flow Cytometry, Small Interfering RNA, Negative Control

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: STAT1 regulates TUBB4A expression at the transcription level. (A) STAT1 mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (B) TUBB4A mRNA levels were measured in A375 and RPMI-7951 cells after STAT1 knockdown via transfection with different siRNA sequences. (C) Specific fragments of the TUBB4A promoter region were cloned into the luciferase reporter plasmids upstream of the firefly luciferase gene. (D) Transcriptional activity of various TUBB4A promoter fragments was analyzed by luciferase reporter assay in 293T cells, with the −1,783 and −1,771 fragments exhibiting the highest activity. (E) STAT1 siRNA-mediated knockdown significantly reduced STAT1 mRNA levels in A375 cells. (F) STAT1 knockdown significantly reduced the luciferase activity of the −1,783 fragment of the TUBB4A promoter, but not the −1,771 fragment. (G) Chromatin immunoprecipitation assays were performed in A375 and RPMI-7951 cells targeting the −1,783 binding site in the TUBB4A promoter region. Quantitative PCR provided evidence of STAT1 binding to this region. Genomic DNA input was set to 100%. **P<0.01 vs. si-NC; ## P<0.01 vs. PGL3; && P<0.01 vs. IgG. STAT1, signal transducer and activator of transcription 1; siRNA, small interfering RNA; si-NC, negative control siRNA; si-STAT1, siRNA targeting STAT1; si-STAT1-1, siRNA targeting STAT1 sequence 1; si-STAT1-2, siRNA targeting STAT1 sequence 2; TUBB4A, tubulin β4A; PGL3, promoter-gluc luciferase 3; LUC, firefly luciferase gene.

Article Snippet: The melanoma cell lines A2058 (cat. no. CRL-3601), SK-MEL-1 (cat. no. HTB-67), A375 (cat. no. CRL-1619) and RPMI-7951 (cat. no. HTB-66), as well as normal human epidermal melanocytes (HEM; cat. no. PCS-200-013) and 293T cells (cat. no. CRL-3216) were obtained from American Type Culture Collection.

Techniques: Expressing, Knockdown, Transfection, Clone Assay, Luciferase, Activity Assay, Reporter Assay, Chromatin Immunoprecipitation, Binding Assay, Real-time Polymerase Chain Reaction, Small Interfering RNA, Negative Control, Sequencing

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression mitigates the effects of STAT1 knockdown on cell viability and proliferation. TUBB4A mRNA levels were measured in (A) A375 and (B) RPMI-7951 cells following TUBB4A overexpression mediated by a lentiviral vector. (C) TUBB4A protein expression was analyzed after its overexpression. Combined STAT1 knockdown and TUBB4A overexpression transfections were performed, followed by a western blot analysis of TUBB4A protein levels in (D) A375 and (E) RPMI-7951 cells. Cell viability was assessed via Cell Counting Kit-8 assays following combined STAT1 knockdown and TUBB4A overexpression in (F) A375 and (G) RPMI-7951 cells. (H) Colony formation assays were performed to assess the proliferative capacity of cells after STAT1 knockdown and TUBB4A overexpression. (I) Quantification of colony formation assay results. **P<0.01 vs. Ov-NC; ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; STAT1, signal transducer and activator of transcription 1; si-NC, negative control small interfering RNA; si-STAT1, small interfering RNA targeting STAT1; Ov-NC, negative control lentiviral overexpression vector; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: The melanoma cell lines A2058 (cat. no. CRL-3601), SK-MEL-1 (cat. no. HTB-67), A375 (cat. no. CRL-1619) and RPMI-7951 (cat. no. HTB-66), as well as normal human epidermal melanocytes (HEM; cat. no. PCS-200-013) and 293T cells (cat. no. CRL-3216) were obtained from American Type Culture Collection.

Techniques: Over Expression, Knockdown, Plasmid Preparation, Expressing, Transfection, Western Blot, Cell Counting, Colony Assay, Negative Control, Small Interfering RNA

Journal: Molecular Medicine Reports

Article Title: STAT1 accelerates cutaneous melanoma progression through TUBB4A expression regulation

doi: 10.3892/mmr.2026.13828

Figure Lengend Snippet: TUBB4A overexpression reverses the effects of STAT1 knockdown on apoptosis, migration and tumor growth. (A-D) Apoptosis and migration were evaluated in A375 and RPMI-7951 cells following STAT1 knockdown and TUBB4A overexpression. Magnification, ×200. (E) Representative images of isolated xenograft tumors in mice. Tumor volumes were measured in nude mice injected subcutaneously with 2×10 6 A375 cells that had been subject to STAT1 knockdown and TUBB4A overexpression. (F) Quantification of mouse tumor volumes showed that STAT1 knockdown significantly suppressed tumor growth, whereas TUBB4A overexpression reversed this inhibitory effect. ## P<0.01 vs. si-STAT1. TUBB4A, tubulin β4A; si-NC, negative control small interfering RNA; Ov-TUBB4A, lentiviral vector for TUBB4A overexpression.

Article Snippet: The melanoma cell lines A2058 (cat. no. CRL-3601), SK-MEL-1 (cat. no. HTB-67), A375 (cat. no. CRL-1619) and RPMI-7951 (cat. no. HTB-66), as well as normal human epidermal melanocytes (HEM; cat. no. PCS-200-013) and 293T cells (cat. no. CRL-3216) were obtained from American Type Culture Collection.

Techniques: Over Expression, Knockdown, Migration, Isolation, Injection, Negative Control, Small Interfering RNA, Plasmid Preparation