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Background: : Exosomal RNAs (ExoRNAs) offer valuable insights into their cellular origin. ExoRNA studies were faced with challenges in obtaining sufficient amounts of high-quality RNA. Herein, we aimed to compare three traditional exosome isolation methods to introduce an appropriate strategy to extract RNA from cancer-derived exosomes for further RNA analysis.
Methods: Exosomes were isolated through ultracentrifugation, precipitation kit, and size exclusion column chromatography, and then characterized by dynamic light scattering and transmission electron microscopy, followed by extracting total RNA. The quality and quantity of the extracted RNAs were assessed by a NanoDrop and 2.5% agarose gel electrophoresis.
Results: Extracted exosomes displayed a similar range of size and morphology. We found that polyethylene glycol-precipitation method resulted in a higher RNA yield with a 260/280 ratio of 1.9. The obtained exoRNA appeared as a smear in the agarose gel, indicative of small exoRNAs.
Conclusion: We provide researchers a suitable approach to isolate exosomes based on yield and purity of exoRNA.
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http://dx.doi.org/10.61186/ibj.4129 | DOI Listing |
Exp Cell Res
September 2025
Cancer Biology Laboratory, Dept of Life Sciences, GITAM School of Sciences, GITAM (Deemed to be University), Visakhapatnam-530045, Andhra Pradesh, India. Electronic address:
CD151 is a tetraspanin, abnormally expressed in triple negative breast cancer (TNBC). It is a prominent component of exosomes, facilitating the secretion of proteins that promote metastasis and drug resistance. We have previously demonstrated that silencing the CD151 gene reduces metastasis in TNBC.
View Article and Find Full Text PDFInt Immunopharmacol
September 2025
Department of Animal Science, College of Agricultural, Yanbian University, Yanji 133002, China; Jilin Provincial Key Laboratory of Transgenic Animal and Embryo Engineering, Yanbian University, Yanji 133002, China. Electronic address:
Objective: Long-term administration of dexamethasone (DEX) to treat severe inflammation or autoimmune disorders often result in skeletal muscle atrophy and functional decline. Exosomes facilitate intercellular communication by transferring bioactive molecules, reflecting the characteristics of their tissue of origin. Myostatin-knockout (MSTN) mice exhibit muscle hypertrophy, and their muscle-derived exosomes (KO-EXOs) retain this phenotype.
View Article and Find Full Text PDFLung Cancer
September 2025
Division of Respiratory Diseases, Department of Internal Medicine, The Jikei University School of Medicine, 3-25-8 Nishi-Shimbashi, Minato-ku, Tokyo, Japan; Division of Next-Generation Drug Development Research, Research Center for Medical Sciences, The Jikei University School of Medicine, 3-25-8 Ni
Background: The risk factors associated with treatment resistance to consolidation durvalumab following chemoradiotherapy (CRT) for locally advanced non-small cell lung cancer (NSCLC) have not been well established.
Methods: Extracellular vesicles (EVs) were isolated from the pretreatment serum of 73 patients treated with consolidation durvalumab. Isolation was performed using CD9/CD63 antibodies, and EV proteins were identified using liquid chromatography-tandem mass spectrometry (LC-MS).
JMIR Res Protoc
September 2025
Department of Medical Oncology, Early Phase Unit, Georges-François Leclerc Centre, Dijon, France.
Background: Sarcomas are rare cancer with a heterogeneous group of tumors. They affect both genders across all age groups and present significant heterogeneity, with more than 70 histological subtypes. Despite tailored treatments, the high metastatic potential of sarcomas remains a major factor in poor patient survival, as metastasis is often the leading cause of death.
View Article and Find Full Text PDFJ Proteome Res
September 2025
State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Research Unit of Proteomics & Research and Development of New Drug of Chinese Academy of Medical Sciences, Institute of Lifeomics, Beijing 102206, China.
Hepatocellular carcinoma (HCC) constitutes approximately 90% of liver cancers, yet its early detection remains challenging due to the low sensitivity of current diagnostic methods and the difficulty in identifying minimal cancer cells within the body. This study employed a patient-derived xenograft (PDX) mouse model to screen for biomarkers, leveraging its advantage of low background interference compared to human serum exosome studies. Using a novel microextraction technique, exosomes were isolated from just one microliter of serum from HCC PDX mice, followed by proteomic profiling.
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