miR-365 (microRNA): Potential Biomarker in Oral Squamous Cell Carcinoma Exosomes and Extracellular Vesicles
»
miR-365 (microRNA): Potential Biomarker in Oral Squamous Cell Carcinoma Exosomes and Extracellular Vesicles
Introduction: miR-365 is a non-coding microRNA that regulates transcription and have been shown to promote metastasis in oncogenes and some types of cancer, while suppressing the effect on others. Many microRNAs is produced and then exported extracellular in exosomes, the small extracellular vesicles ranging from 30 to 100 nm are found in eukaryotic fluid and facilitate various cellular functions.
Exosomes and extracellular vesicles produced by many types of cells, including cancer cells mouth-though no study to date has evaluated miR-365 and oral cancer exosomes or extracellular vesicles. Based on this information, the question of our research is to evaluate whether the cancer of the mouth produces extracellular exosomes or vesicles containing miR-365.
Materials and methods: Two commercially available oral cancer cell lines (SCC25 and CAL27) and normal oral keratinocytes (OKF4) were grown in serum-free media, supplemented with exosome-depleted fetal bovine serum. extracellular vesicles and exosomes then isolated using Invitrogen Total exosome RNA and protein isolation kit for processing using HSA-miR-365a-5p microRNA qPCR assay kits.
Results: RNA was isolated from exosome-depleted supernatant from each cell line-SCC9, SCC15, SCC25, and CAL27 (oral squamous cell carcinoma) and OKF4 (oral epithelial cell lines). the relative concentration of RNA were similar among each cell line, which is not significant, p = 0.233. RNA quality was established by A260: A280 absorbance using a NanoDrop, expressing the purity ranging from 1.73 to 1.86. Expression of miR-16 microRNA is used to confirm the presence of exosomes extracted and extracellular vesicles. The presence of miR-365 was later confirmed and normalized to miR-16 expression, which indicates the rate of increase of miR-365 in both CAL27 and SCC25. In addition, the normalization relative quantity (RQ) for miR-365 exhibited a greater variation between SCC25 (1.382 to 4.363) of cells CAL27 (1.248 to 1.536).
Conclusion: These results confirm that miR-365 was not only expressed in cancer cell lines of the mouth, but also subsequently exported to exosomes and extracellular vesicles derived from this culture. This data can help to contextualize this microRNA potential to contribute to the phenotype and behavior of oral cancers that express this microRNA. Future research will begin to investigate the potential mechanisms and pathways and to determine whether miR-365 may be useful as a biomarker of cancer of the mouth for saliva or fluid biopsy.
miR-365 (microRNA): Potential Biomarker in Oral Squamous Cell Carcinoma Exosomes and Extracellular Vesicles
Exosomal Circulating microRNAs as potential novel biomarker detection in pancreatic cancer
Circulating microRNAs exosomal (ex-miRNA) is reflective of the characteristics of the tumor and valuable biomarkers in a variety of tumor types. In addition, miRNAs serve an important role in tumor progression and metastasis.
This study aims to determine the outstanding ex-miRNA-21 and miRNA-210 as a new biomarker for patients with pancreatic cancer (PC). For this purpose, the former serum miRNAs extracted from the serum of patients with PC (n = 30) and chronic pancreatitis (CP) (n = 10) using the RNA isolation kit.
Description: SDS-Blue™ is an innovative patented formula, based on Coomassie blue, that comes in a convenient ready to use format for staining proteins in SDS-PAGE (sodium dodecyl sulphate–polyacrylamide gel electrophoresis). The formulation of SDS-Blue™ provides numerous advantages compared to the classic Coomassie staining or to other similar protein stains. SDS-Blue™ provides higher sensitivity, virtually no background and eliminates the need for destaining of the gel due to its high specificity and affinity to bind to protein only. Not only does SDS-Blue™ yield clear and sharp bands, but it also contains no methanol and acetic acid, making it non-hazardous, safe to handle and friendly to the environment when disposed of. Two other advantages that make SDS-Blue™ the better option is that it is not light sensitive and can be stored at ambient temperature for 24 months. And this provides a considerable convenience, especially to laboratories that need and keep big amount of protein staining solutions – no more jammed refrigerators, you can keep SDS-Blue™ wherever it is most convenient for You!
Thrombin Plasma prep for Exosome precipitation (500 ul at 500U/mL)
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.
Description: Can be used for various proteomics studies in both normal and pathological cases. It is an excellent control and suitable for educational purposes. This product is prepared from whole tissue homogenates and has undergone SDS-PAGE quality control analysis. The protein is stored in a buffer with protease inhibitor cocktail fo prevent degradation.
Description: The anti-IgY beads were made by cross-linking of bovine IgG anti-IgY antibodies to Protein A/G agarose beads. IgY is the original designation of chicken IgG like immunoglobulin.
Description: Recombinant Human IL-4 produced in E.Coli is a single, non-glycosylated polypeptide chain containing 130 amino acids and having a molecular mass of 15000 Dalton. The rHuIL-4 is purified by proprietary chromatographic techniques.
Description: Recombinant human interleukin-2 is a sterile protein product for injection. rHuIL-2 is produced by recombinant DNA technology using Yeast. It is a highly purified protein containing 133 amino acids, with cysteine mutated to alanine at 125 amino acid position, and has a molecular weight of approximately 15.4kD, non-glycosylated.
Description: Recombinant human interleukin-2 is a sterile protein product for injection. rHuIL-2 is produced by recombinant DNA technology using Yeast. It is a highly purified protein containing 133 amino acids, with cysteine mutated to alanine at 125 amino acid position, and has a molecular weight of approximately 15.4kD, non-glycosylated.
Description: Recombinant human GM-CSF produced in E.coli is a single, non-glycosylated, polypeptide chain containing 127 amino acids, two pairs of disulfide bonds and having a molecular mass of approximately 14.5kD.
Description: Monkeypox virus is the virus that causes the disease monkeypox in both humans and animals. Monkeypox virus is an Orthopoxvirus, a genus of the family Poxviridae that contains other viral species that target mammals. The virus is mainly found in tropical rainforest regions of central and West Africa. The primary route of infection is thought to be contact with the infected animals or their bodily fluids. The genome is not segmented and contains a single molecule of linear double-stranded DNA, 185000 nucleotides long. The Monkeypox Virus real time PCR Kit contains a specific ready-to-use system for the detection of the Monkeypox Virusthrough polymerase chain reaction (PCR) in the real-time PCR system. The master contains reagents and enzymes for the specific amplification of the Monkeypox Virus DNA. Fluorescence is emitted and measured by the real time systems ́ optical unit during the PCR. The detection of amplified Monkeypox Virus DNA fragment is performed in fluorimeter channel 530nm with the fluorescent quencher BHQ1. DNA extraction buffer is available in the kit and serum or lesion exudate samples are used for the extraction of the DNA. In addition, the kit contains a system to identify possible PCR inhibition by measuring the 560nm fluorescence of the internal control (IC). An external positive control defined as 1×10^7 copies/ml is supplied which allow the determination of the gene load.
Description: Monkeypox virus is the virus that causes the disease monkeypox in both humans and animals. Monkeypox virus is an Orthopoxvirus, a genus of the family Poxviridae that contains other viral species that target mammals. The virus is mainly found in tropical rainforest regions of central and West Africa. The primary route of infection is thought to be contact with the infected animals or their bodily fluids.The genome is not segmented and contains a single molecule of linear double-stranded DNA, 185000 nucleotides long.The Monkeypox Virus real time PCR Kit contains a specific ready-to-use system for the detection of the Monkeypox Virusthrough polymerase chain reaction (PCR) in the real-time PCR system. The master contains reagents and enzymes for the specific amplification of theMonkeypox VirusDNA. Fluorescence is emitted and measured by the real time systems ́ optical unit during the PCR. The detection of amplified Monkeypox Virus DNA fragment is performed in fluorimeter channelFAM with the fluorescent quencher BHQ1. DNA extraction buffer is available in the kit and serum or lesion exudate samples are used for the extraction of the DNA. In addition, the kit contains a system to identify possible PCR inhibition by measuring the HEX/VIC/JOE fluorescence of the internal control (IC). An external positive control defined as 1×107copies/ml is supplied which allow the determination of the gene load.
To exosome identified in serum, transmission electron micrographs were used to determine the crystal structure, western blotting was used to identify markers exosomal, and NanoSight used to characterize the nanoparticles.
