Journal: Oncology Letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: HOXB4 promotes STARD13 expression by directly binding to the STARD13 promoter. (A and B) The infection efficiencies of Lenti-HOXB4-CDS and Lenti-HOXB4-shRNA were detected by qRT-PCR and western blot analyses in MCF-7 and MDA-MB-231 cells. (C and D) STARD13 mRNA and protein levels were determined in cells with stable expression of HOXB4-CDS or knockdown of HOXB4. (E) The promoter activity of STARD13 was detected by luciferase reporter assay in 293T cells. (F and G) qRT-PCR was used to measure the enrichment of STARD13 promoter sequences in the immunoprecipitated complexes using promoter-specific primer sets in MCF-7 and MDA-MB-231 cells with stable expression of HOXB4. Data are presented as means ± SDs; n=3, **P<0.01 vs. the control.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Expressing, Binding Assay, Infection, shRNA, Quantitative RT-PCR, Western Blot, Knockdown, Activity Assay, Luciferase, Reporter Assay, Immunoprecipitation, Control

Journal: Oncology Letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: HOXB4 inhibites breast cancer cell migration in a STARD13-dependent manner. (A-D) Transwell migration assays were used to detect the migration ability of MCF-7 and MDA-MB-231 cells. Overexpression of HOXB4 in MDA-MB-231 cells or knockdown of HOXB4 in MCF-7 cells inhibited or promoted migration, respectively, and this effect was attenuated or reversed by co-infection with Ad-STARD13-shRNA or Ad-STARD13-CDS. Data are presented as means ± SDs; n=3, **P<0.01 vs. the control.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Migration, Over Expression, Knockdown, Infection, shRNA, Control

Journal: Oncology Letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: HOXB4 inhibites breast cancer adhesion ability and the EMT in a STARD13-dependent manner. (A and B) Cell adhesion assay of MDA-MB-231 cells with HOXB4 overexpression and MCF-7 cells with HOXB4 knockdown. (C-F) qRT-PCR and western blotting were used to detect the expression levels of EMT markers (Vimentin and E-cadherin) in MDA-MB-231 cells co-treated with Lenti-HOXB4-CDS and Ad-STARD13-shRNA and MCF-7 cells co-treated with Lenti-HOXB4-shRNA and Ad-STARD13-CDS. (G) Western blotting was used to detect E-cadherin protein levels in MCF-7 cells co-treated with Lenti-HOXB4-shRNA and Y-27632. Data are presented as means ± SD. n=3, **P<0.01.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Cell Adhesion Assay, Over Expression, Knockdown, Quantitative RT-PCR, Western Blot, Expressing, shRNA

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 1. HOXB4 and STARD13 expression levels are downregulated in breast cancer tissues and cells. (A and B) HOXB4 and STARD13 mRNA levels were detected in breast cancer with metastasis (n=20) or not (n=27) and normal tissues (n=27) by qRT‑PCR. (C and D) HOXB4 and STARD13 protein levels were examined in breast cancer and normal tissues by immunohistochemistry analysis. (E) qRT‑PCR results showed that HOXB4 and STARD13 expression levels were positively correlated. Data are presented as means ± SD; n=3, *P<0.05, **P<0.01 vs. the control.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Expressing, Immunohistochemistry, Control

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 3. HOXB4 and STARD13 expression levels are positively correlated with patients survival. (A and B) TCGA data were used to test the correlations between survival and HOXB4/STARD13‑expression levels. (C and D) HOXB4 mRNA and protein levels were determined in normal breast epithelial cells (MCF‑10A) and breast cancer cell lines (MCF‑7, MDA‑MB‑435 and MDA‑MB‑231). Data are presented as means ± SD; n=3, *P<0.05, **P<0.01 vs. the control.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Expressing, Control

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 2. HOXB4 and STARD13 expression levels are positively correlated. (A and B) TCGA data set was used to test the correlation between STARD13 and HOXB4 expression levels in two different clinical samples.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Expressing

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 4. HOXB4 promotes STARD13 expression by directly binding to the STARD13 promoter. (A and B) The infection efficiencies of Lenti‑HOXB4‑CDS and Lenti‑HOXB4‑shRNA were detected by qRT‑PCR and western blot analyses in MCF‑7 and MDA‑MB‑231 cells. (C and D) STARD13 mRNA and protein levels were determined in cells with stable expression of HOXB4‑CDS or knockdown of HOXB4. (E) The promoter activity of STARD13 was detected by luciferase reporter assay in 293T cells. (F and G) qRT‑PCR was used to measure the enrichment of STARD13 promoter sequences in the immunoprecipitated complexes using promoter‑specific primer sets in MCF‑7 and MDA‑MB‑231 cells with stable expression of HOXB4. Data are presented as means ± SDs; n=3, **P<0.01 vs. the control.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Expressing, Binding Assay, Infection, Western Blot, Knockdown, Activity Assay, Luciferase, Reporter Assay, Immunoprecipitation, Control

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 6. HOXB4 inhibites breast cancer adhesion ability and the EMT in a STARD13‑dependent manner. (A and B) Cell adhesion assay of MDA‑MB‑231 cells with HOXB4 overexpression and MCF‑7 cells with HOXB4 knockdown. (C‑F) qRT‑PCR and western blotting were used to detect the expression levels of EMT markers (Vimentin and E‑cadherin) in MDA‑MB‑231 cells co‑treated with Lenti‑HOXB4‑CDS and Ad‑STARD13‑shRNA and MCF‑7 cells co‑treated with Lenti‑HOXB4‑shRNA and Ad‑STARD13‑CDS. (G) Western blotting was used to detect E‑cadherin protein levels in MCF‑7 cells co‑treated with Lenti‑HOXB4‑shRNA and Y‑27632. Data are presented as means ± SD. n=3, **P<0.01.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Cell Adhesion Assay, Over Expression, Knockdown, Western Blot, Expressing

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 5. HOXB4 inhibites breast cancer cell migration in a STARD13‑dependent manner. (A‑D) Transwell migration assays were used to detect the migration ability of MCF‑7 and MDA‑MB‑231 cells. Overexpression of HOXB4 in MDA‑MB‑231 cells or knockdown of HOXB4 in MCF‑7 cells inhibited or promoted migration, respectively, and this effect was attenuated or reversed by co‑infection with Ad‑STARD13‑shRNA or Ad‑STARD13‑CDS. Data are presented as means ± SDs; n=3, **P<0.01 vs. the control.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Migration, Over Expression, Knockdown, Control

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 7. Overexpression of HOXB4 enhanced doxorubicin‑mediated downregulation of cell viability in breast cancer cells. (A and B) HOXB4 mRNA and protein levels were detected in MCF‑7 and MCF‑7‑ADR cells. (C) Cell viability was examined using the MTT assays in MCF‑7‑ADR cells treated with doxorubicin with or without Ad‑HOXB4‑CDS. (D) Cell viability of MCF‑7 cells treated with doxorubicin with or without Ad‑HOXB4‑CDS. (E and F) Cell viability in MCF‑7 and MDA‑MB‑231 cells with HOXB4 knockdown or overexpression, respectively. **P<0.01 vs. the control.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Over Expression, Knockdown, Control

Journal: Oncology letters

Article Title: Homeobox B4 inhibits breast cancer cell migration by directly binding to StAR-related lipid transfer domain protein 13.

doi: 10.3892/ol.2017.6825

Figure Lengend Snippet: Figure 8. Overexpression of HOXB4 enhanced doxorubicin‑mediated downregulation of cell apoptosis in breast cancer cells. (A‑C) Apoptosis assays was assessed in MCF‑7, MCF‑10Aand MDA‑MB‑231 cells with or without HOXB4 overexpression. (D) Bcl‑2 and Bax protein levels were determined by western blotting in MCF‑7 cells treated with Lenti‑HOXB4‑CDS with or without doxorubicin. (E) Bcl‑2 and Bax protein levels were determined by western blotting in MCF‑7 cells with or without HOXB4 knockdown. Data are presented as means ± SD. n=3, *P<0.05, **P<0.01.

Article Snippet: Lentivirus short hairpin(sh)RNA against human HOXB4 and a scramble non-targeting shRNA (sc-38692) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA), and inserted into pLKO.1.

Techniques: Over Expression, Western Blot, Knockdown