Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: The INHBA is upregulated in BC cells and facilitates cell growth. (a) The INHBA overexpression in four BC cell lines was measured by RT-qPCR and Western blot. β-actin was used as the loading control. (b) The efficiency of INHBA knockdown in BT549 cells by sh-INHBA#1/2 or sh-NC vectors was assessed by RT-qPCR and Western blot. GAPDH was used as a loading control in Western blot. (c) CCK-8 assay and (d) colony formation assay were performed to examine the cell proliferation of BT549 cells after the transfection of shRNA. (e) The migration of transfected cells was examined by wound healing assay. (f) The invasion ability of transfected cells was determined by transwell assays (×200 magnification). Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing sh-INHBA (OriGene, Beijing, China) or negative control sh-NC (OriGene, Beijing, China) was transfected to BT549 cells. shRNA sequence targeting human INHBA were shown as below: INHBA#1: 5ʹ-CCATGTCCATGTTGTACTA-3ʹ, INHBA#2: 5ʹ-GCAACAAATTGATGAGCAA-3ʹ, and control sequence: 5ʹ-CAACAAGATGAAGAGCACCAA-3ʹ. qRT-PCR was performed to confirm the overexpression or knockdown efficiency 48 hours after transfection.

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, CCK-8 Assay, Colony Assay, Transfection, shRNA, Migration, Wound Healing Assay

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: INHBA induces the EMT in BC cells. (a) Western blot analysis of the EMT-related proteins after knockdown of INHBA in BT549 cells (left) and overexpression in MCF-7 cells (right). (b) RT-qPCR analysis of the EMT-related genes after knockdown of INHBA in BT549 cells (left) and overexpression in MCF-7 cells (right). GAPDH was used as a loading control in Western blot. Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing sh-INHBA (OriGene, Beijing, China) or negative control sh-NC (OriGene, Beijing, China) was transfected to BT549 cells. shRNA sequence targeting human INHBA were shown as below: INHBA#1: 5ʹ-CCATGTCCATGTTGTACTA-3ʹ, INHBA#2: 5ʹ-GCAACAAATTGATGAGCAA-3ʹ, and control sequence: 5ʹ-CAACAAGATGAAGAGCACCAA-3ʹ. qRT-PCR was performed to confirm the overexpression or knockdown efficiency 48 hours after transfection.

Techniques: Western Blot, Over Expression, Quantitative RT-PCR

Journal: Bioengineered

Article Title: Inhibin β-A (INHBA) induces epithelial–mesenchymal transition and accelerates the motility of breast cancer cells by activating the TGF-β signaling pathway

doi: 10.1080/21655979.2021.1957754

Figure Lengend Snippet: INHBA overexpression activates the TGF-β signaling pathway, which is suppressed by TGF-β inhibitor. (a) The protein level of TGF-β1, Smad2, p-Smad2, Smad3, p-Smad3 and VEGF-A in BT549 (INHBA knockdown) and MCF-7 cells (INHBA overexpression). GAPDH was used as loading control in Western blot. (b) The protein level of Smad2, p-Smad2, Smad3, p-Smad3, TGF-β1, INHBA in MCT-7 cells treated by TGF-β inhibitor (SB-431,542). 10 µM SB-431,542 was used as final concentration in the medium. DMSO solvent was also used as a negative control. (c) The cell migration (upper two) and invasion (lower two) analyses for MCF-7 cells treated with (or without) TGF-β inhibitor (×200 magnification). (d) Western blot analysis of EMT-related genes in MCF-7 cells transfected with (or without) INHBA expression vector, along with (or without) SB-431,542. Data are summary of 3 independent experiments (n = 3). The error bars are defined as s.d. *, P < 0.05, **, P < 0.01, and ***, P < 0.001

Article Snippet: The plasmid containing sh-INHBA (OriGene, Beijing, China) or negative control sh-NC (OriGene, Beijing, China) was transfected to BT549 cells. shRNA sequence targeting human INHBA were shown as below: INHBA#1: 5ʹ-CCATGTCCATGTTGTACTA-3ʹ, INHBA#2: 5ʹ-GCAACAAATTGATGAGCAA-3ʹ, and control sequence: 5ʹ-CAACAAGATGAAGAGCACCAA-3ʹ. qRT-PCR was performed to confirm the overexpression or knockdown efficiency 48 hours after transfection.

Techniques: Over Expression, Western Blot, Concentration Assay, Negative Control, Migration, Transfection, Expressing, Plasmid Preparation

Journal: Oncotarget

Article Title: High-throughput RNAi screening for novel modulators of vimentin expression identifies MTHFD2 as a regulator of breast cancer cell migration and invasion

doi:

Figure Lengend Snippet: A: The effect of MTHFD2 siRNAs on MDA-MB-231(SA) and BT-549 cell motility in wound healing experiment. Cells were automatically imaged once every hour after wound scratching. Wound closure effect was calculated as wound confluence which the cells gained in twelve hours. B: MDA-MB-231(SA) cells transfected with either vimentin targeting siRNA (siVIM), MTHFD2 targeting siRNA (siMTHFD2) or negative control siRNA (siScr), allowed to invade into Matrigel matrix for 4 days and imaged. Shown are the quantifications of two replicate experiments (imaged at 5-10 different positions; ***p < 0.005) and representative images. The dashed lines indicate the top of the Matrigel matrix.

Article Snippet: BT-549 cells were obtained from Cell Lines Service (Eppelheim, Germany).

Techniques: Transfection, Negative Control

Journal: Scientific Reports

Article Title: TRAF6 as a potential target in advanced breast cancer: a systematic review, meta-analysis, and bioinformatics validation

doi: 10.1038/s41598-023-31557-0

Figure Lengend Snippet: Summary of meta-analysis of included studies showing significant association of in vitro breast cancer cell behaviour and modulation of TRAF2/4/6.

Article Snippet: , Proliferation , Pharmacological inhibition , BT549 (4) MCF7 (4) MDA-MB-231 (8) , − 5.78 [− 12.04, 0.47] − 9.50 [− 18.82, − 0.19] − 5.62 [− 10.34, − 0.90] , − 21.46 [− 30.40, − 12.51] , Mean Difference (IV, Random, 95% CI) , Tau 2 = 321.80; Chi 2 = 677.01, df = 15 (P < 0.00001); I 2 = 98% , Z = 4.70 (P < 0.00001).

Techniques: In Vitro, Migration, Inhibition

Journal: Scientific Reports

Article Title: TRAF6 as a potential target in advanced breast cancer: a systematic review, meta-analysis, and bioinformatics validation

doi: 10.1038/s41598-023-31557-0

Figure Lengend Snippet: Summary of meta-analysis of included studies showing significant association of in vivo tumour burden and overt metastasis with modulation of TRAF2/4/6.

Article Snippet: , Proliferation , Pharmacological inhibition , BT549 (4) MCF7 (4) MDA-MB-231 (8) , − 5.78 [− 12.04, 0.47] − 9.50 [− 18.82, − 0.19] − 5.62 [− 10.34, − 0.90] , − 21.46 [− 30.40, − 12.51] , Mean Difference (IV, Random, 95% CI) , Tau 2 = 321.80; Chi 2 = 677.01, df = 15 (P < 0.00001); I 2 = 98% , Z = 4.70 (P < 0.00001).

Techniques: In Vivo, Inhibition