Journal: Cancer Cell International

Article Title: Circulating extracellular vesicle PTPRO methylation: an exploratory biomarker for minimally invasive diagnosis of early-stage lung adenocarcinoma

doi: 10.1186/s12935-025-04127-9

Figure Lengend Snippet: Methylation analysis of PTPRO in LUAD cell lines and LUAD cells derived sEVs. (A) The mRNA expression levels of PTPRO were determined by RT-qPCR in the normal lung epithelial cell line HBE and LUAD cell lines A549, NCI-H460, and PC-9. (B-C) PTPRO methylation status in HBE and the LUAD cell lines A549, NCI-H460, and PC-9 was assessed using MSP and q-MSP assays. (D) Representative TEM images of sEVs purified from the conditioned media of HBE and LUAD cell lines (A549, NCI-H460, and PC-9). Scale bar: 100 nm. (E) The concentration and size distribution of sEVs from HBE and LUAD cell lines (A549, NCI-H460, and PC-9) were evaluated using NTA. (F) Expression of sEV markers (TSG101, Alix, CD63, and Calnexin) and PTPRO in sEVs purified from HBE and LUAD cell lines (A549, NCI-H460, and PC-9) was detected by immunoblotting. (G) The mRNA expression levels of PTPRO in sEVs purified from the conditioned media of HBE and LUAD cell lines (A549, NCI-H460, and PC-9) were measured by RT-qPCR. (H-I) MSP and q-MSP analyses of purified sEVs from HBE and LUAD cell lines (A549, NCI-H460, and PC-9). H 2 O was used as a negative control. Error bars indicate SEM. *** P < 0.001 by one-way ANOVA with Dunnett's multiple comparisons test.

Article Snippet: Human LUAD cell lines (A549, PC-9, and NCI-H460) and normal human bronchial epithelial cells (HBE) were obtained from the American Type Culture Collection (ATCC; Manassas, VA, USA).

Techniques: Methylation, Derivative Assay, Expressing, Quantitative RT-PCR, Purification, Concentration Assay, Western Blot, Negative Control