Journal: Journal of nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway.

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: Fig. 1 Schematic illustration of the therapeutic system based on EVs used for the delivery of nucleic acid drugs targeting gene mutation sites for the precise treatment of CRC with the BRAFV600E mutation. siBRAFV600E was transfected into HEK293T cells via a lentiviral vector to stably produce EVs carrying siBRAFV600E. After treatment with these EVs- siBRAFV600E, the BRAF-MEK1/2-ERK1/2 pathway was inhibited in CRC cells. Subcutaneous xenograft tumor models and metastasis models of CRC cell lines were used to validate the therapeutic effects and toxicity of the EVs-siBRAFV600E. The efficacy of the EVs- siBRAFV600E was further confirmed via PDX models

Article Snippet: The primary antibodies used were against BRAFV600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Mutagenesis, Transfection, Plasmid Preparation, Stable Transfection

Journal: Journal of nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway.

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: Fig. 2 BRAF mutation is a poor prognostic factor in CRC patients. (A) K-M survival curves of patients with BRAF mutation and wild-type CRC. (B) Multivari ate COX regression model analysis of gender, stage and BRAF mutation status in CRC patients. (C) K-M survival curve of BRAF expression level in BRAFMT CRC. (D) Representative IHC images of BRAFV600E and KI-67 in CRC tissues. The scale bar represents 10 μm for the 10x objective and 5 μm for the 20x objective. (E) KI67 positivity rate in CRC tissues with BRAFV600E or BRAFWT. The data are reported as the means ± SDs of the experiments (n = 3). Two-tailed Student’s t-tests, **p < 0.01

Article Snippet: The primary antibodies used were against BRAFV600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: Mutagenesis, Expressing, Two Tailed Test

Journal: Journal of nanobiotechnology

Article Title: Extracellular vesicle-mediated gene therapy targets BRAF V600E -mutant colorectal cancer by inhibiting the MEK1/2-ERK1/2 pathway.

doi: 10.1186/s12951-025-03205-4

Figure Lengend Snippet: Fig. 7 EVs-siBRAFV600E inhibit BRAFV600E CRC in vivo. (A) Animal model handling protocol. (B) Subcutaneous tumor volume changes. Tumor volume measurements were performed every 3 days. (C) Images of the subcutaneous tumors in the three groups. (D) The weights of the subcutaneous tumors. (E) Immunohistochemistry images of BRAF and p-ERK1/2. Scale bar: 50 μm. (F)~(I) WB detection of BRAF expression and MEK1/2-ERK1/2 phosphorylation levels in subcutaneous tumor tissues. (J) Bioluminescence images of lung metastases. (K) Bioluminescence images of liver metastases. Each group of animals was treated with PBS, EVs-NC or EVs-siBRAFV600E. Each mouse was injected with 20 µg of extracellular vesicles per injection. Subcutaneous tumors: n = 5 per group. Lung metastasis and liver metastasis: n = 4 per group. The data are reported as the means ± SDs of the experiments. Two-way ANOVA for (B), and one-way ANOVA followed by Tukey test multiple comparisons for the others, *p < 0.05, **p < 0.01 and ***p < 0.001

Article Snippet: The primary antibodies used were against BRAFV600E (ZA-0668, ZSGB-BO, China), KI67 (27309-1-AP, Proteintech), BRAF (ab33899, Abcam), and phospho-p44/42 MAPK (p-ERK1/2) (4370s, Cell Signaling Technology).

Techniques: In Vivo, Animal Model, Immunohistochemistry, Expressing, Phospho-proteomics, Injection