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.
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.
 Exosome precipitation Solution |
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| EPStep | ImmunoStep | 12 ml | EUR 277.5 |
) ExoQuick Plasma prep and Exosome precipitation kit (5 ml ExoQuick plus 500 ul Thrombin at 500U/mL) |
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| EXOQ5TM-1 | SBI | 75 reactions | EUR 843 |
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 Exosome precipitation from
Plasma + Thrombin |
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| EPStep-T | ImmunoStep | 5 ml | EUR 336 |
 ExoQuick Exosome RNA Column Purification kit |
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| EQ808A-1 | SBI | 20 preps | EUR 402 |
 Exosome precipitation from
Plasma and Serum |
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| EPStep-PS | ImmunoStep | 5 ml | EUR 297 |
 ExoQuick-LP for lipoprotein pre-clear & exosome isolation |
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| EXOLP5A-1 | SBI | 5 reactions | EUR 533 |
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) PEG-it Virus Precipitation Solution (250 mL aliquot) |
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| LV825A-1 | SBI | 250 ml | EUR 777 |
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) PEG-it Virus Precipitation Solution (100 mL aliquot) |
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| LV810A-1 | SBI | 100 ml | EUR 382 |
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) ExoQuick-CG cGMP-grade Exosome Isolation reagent (50 ml) |
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| EXOCG50A-1 | SBI | 50 ml | EUR 1181 |
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 ExoQuick PLUS Exosome Purification Kit - for serum and plasma |
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| EQPL10A-1 | SBI | 10 reactions | EUR 579 |
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 ExoQuick-TC PLUS Exosome Purification Kit - for tissue culture media |
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| EQPL10TC-1 | SBI | 10 reactions | EUR 579 |
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) ExoQuick Exosome Isolation and RNA Purification kit (for Serum and Plasma) |
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| EQ806A-1 | SBI | 20 preps | EUR 554 |
) ExoQuick Exosome Isolation and RNA Purification kit (for Tissue Culture Media) |
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| EQ806TC-1 | SBI | 10 preps | EUR 554 |
) SeraMir Exosome RNA Purification kit (5ml ExoQuick and 20 exoRNA columns) |
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| RA806A-1 | SBI | 20 preps | EUR 554 |
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) Thrombin Plasma prep for Exosome precipitation (500 ul at 500U/mL) |
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| TMEXO-1 | SBI | 100 reactions | EUR 583 |
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) Exosome isolation Solution - Serum/ Plasma (easy version of P101, 95% pure exosome) |
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| P101EZ | 101Bio | NULL | EUR 0 |
 ExoQuick TC 10ml |
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| EXOTC10A-1 | SBI | 10 ml | EUR 399 |
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 ExoQuick TC 50ml |
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| EXOTC50A-1 | SBI | 50 ml | EUR 1222 |
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 TMB for Precipitation |
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| 42R-TB100x | Fitzgerald | 1000 ml | EUR 538 |
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Description: High sensitivity TMB Substrate for Precipitation assays |
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 DNA Precipitation Buffer |
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| D106 | Cygnus Technologies | 55 ml | EUR 175 |
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Description: DNA Precipitation Buffer by Cygnus Technologies is available in Europe via Gentaur. |
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) Exosome isolation Solution - Cell Media/ Urine (easy version of P100, 95% pure exosome) |
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| P100EZ | 101Bio | NULL | EUR 0 |
 sample) Exosome isolation Solution - Serum/ Plasma (easy version of P101, 95% pure exosome) sample |
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| P101EZ--sample | 101Bio | NULL | EUR 0 |
 sample) Exosome isolation Solution - Cell Media/ Urine (easy version of P100, 95% pure exosome) sample |
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| P100EZ-sample | 101Bio | NULL | EUR 0 |
 10 preps) SeraMir Exosome RNA Purification kit (10ml ExoQuick-TC and 10 exoRNA columns) 10 preps |
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| RA806TC-1 | SBI | 10 preps | EUR 554 |
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 Chicken IgY precipitation reagent |
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| 85R-1010 | Fitzgerald | 4 ml | EUR 554 |
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Description: Chicken IgY precipitation reagent for use in immunoassays |
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 PEG Virus Precipitation Kit |
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| K904-200 | Biovision | EUR 1001 | |
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PEG Virus Precipitation Kit |
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| K904-50 | Biovision | EUR 376 | |
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PEG Virus Precipitation Kit |
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| K2190-200 | ApexBio | 200 preparations | EUR 1059 |
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PEG Virus Precipitation Kit |
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| K2190-50 | ApexBio | 50 preparations | EUR 405 |
 50ML Protein Precipitation Kit |
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| NAT1254 | National Diagnostics | 50ML | EUR 225 |
) SeraMir Exosome RNA Amplification kit from Media and Urine (10ml ExoQuick-TC and 10 exoRNA columns) |
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| RA800TC-1 | SBI | 10 reactions | EUR 659 |
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 101Bio High Efficiency Protein Precipitation kit |
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| P5W6 | 101Bio | NULL | EUR 0 |
 In Vitro SDS-K+ Precipitation Kit |
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| TG1010-1 | TopoGen | 100 assays | EUR 459 |
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In Vitro SDS-K+ Precipitation Kit |
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| TG1010-2 | TopoGen | 250 assays | EUR 571 |
 In Vivo SDS-K+ Precipitation Kit |
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| TG1011-1 | TopoGen | 100 assays | EUR 302 |
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In Vivo SDS-K+ Precipitation Kit |
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| TG1011-2 | TopoGen | 250 assays | EUR 414 |
 Psoriasis Exosome |
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| P141-PS | 101Bio | NULL | EUR 0 |
 Sodium Pyruvate Solution, 100 mM Solution |
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| CCM1753-100 | BBI Biotech | 100 mL | EUR 60.84 |
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 Lyophilized Exosome Standards |
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| ExoA375 | ImmunoStep | 100 μg | EUR 303.5 |
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Lyophilized Exosome Standards |
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| ExoHT29 | ImmunoStep | 100 μg | EUR 303.5 |
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Lyophilized Exosome Standards |
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| ExoMCF7 | ImmunoStep | 100 μg | EUR 303.5 |
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Lyophilized Exosome Standards |
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| ExoPC3 | ImmunoStep | 100 μg | EUR 303.5 |
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Lyophilized Exosome Standards |
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| ExoPLASMA | ImmunoStep | 100 μg | EUR 303.5 |
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Lyophilized Exosome Standards |
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| ExoUrine | ImmunoStep | 100 μg | EUR 303.5 |
 Leukemia - ALL Exosome |
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| P141-ALL | 101Bio | NULL | EUR 0 |
 Leukemia - AML Exosome |
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| P141-AML | 101Bio | NULL | EUR 0 |
 Bladder Cancer Exosome |
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| P141-BL | 101Bio | NULL | EUR 0 |
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| P141-BR | 101Bio | NULL | EUR 0 |
 Leukemia - CLL Exosome |
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| P141-CLL | 101Bio | NULL | EUR 0 |
 Leukemia - CML Exosome |
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| P141-CML | 101Bio | NULL | EUR 0 |
 Colorectal Cancer Exosome |
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| P141-CO | 101Bio | NULL | EUR 0 |
 Crohn's Disease Exosome |
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| P141-CR | 101Bio | NULL | EUR 0 |
 Kidney Cancer Exosome |
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| P141-KN | 101Bio | NULL | EUR 0 |
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| P141-LG | 101Bio | NULL | EUR 0 |
 Liver Cancer Exosome |
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| P141-LV | 101Bio | NULL | EUR 0 |
 Nasopharyngeal Cancer Exosome |
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| P141-NA | 101Bio | NULL | EUR 0 |
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| P141-PR | 101Bio | NULL | EUR 0 |
 Rheumatoid Arthritis Exosome |
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| P141-RA | 101Bio | NULL | EUR 0 |
 Thyroid Cancer Exosome |
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| P141-TH | 101Bio | NULL | EUR 0 |
 Psoriasis Exosome RNA |
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| P241-PS | 101Bio | NULL | EUR 0 |
 SDS Solution |
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| 2102-100 | Biovision | EUR 137 | |
 EDTA Solution |
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| 2103-100 | Biovision | EUR 137 | |
 Denaturing solution |
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| DE540 | BBI Biotech | 1L | EUR 61.31 |
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 TRIMMING SOLUTION |
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| 99-676-CM | CORNING | 1 L /pk | EUR 88 |
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Description: Specialty Media; Islet products |
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 AlphaBioCoat Solution |
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| AC001 | Neuromics | 20 ml | EUR 271 |
 Borax Solution |
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| BOR015 | ScyTek Laboratories | 15 ml | EUR 56 |
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Borax Solution |
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| BOR125 | ScyTek Laboratories | 125 ml | EUR 79 |
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Borax Solution |
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| BOR500 | ScyTek Laboratories | 500 ml | EUR 119 |
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Borax Solution |
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| BOR999 | ScyTek Laboratories | 1000 ml | EUR 183 |
 Stop Solution |
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| F006 | Cygnus Technologies | 12 ml | EUR 180 |
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Description: Stop Solution by Cygnus Technologies is available in Europe via Gentaur. |
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 Prehybridization Solution |
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| K2191050-1 | Biochain | 5 ml | EUR 137 |
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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. |
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 Hybridization Solution |
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| K2191050-2 | Biochain | 5 ml | EUR 157 |
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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. |
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 Blocking Solution |
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| K2191050-8 | Biochain | 250 ul | EUR 137 |
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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. |
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 Orcein Solution |
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| OAA125 | ScyTek Laboratories | 125 ml | EUR 66 |
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Orcein Solution |
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| OAA500 | ScyTek Laboratories | 500 ml | EUR 109 |
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Orcein Solution |
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| OAA999 | ScyTek Laboratories | 1000 ml | EUR 148 |
 Methenamine Solution |
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| MET125 | ScyTek Laboratories | 125 ml | EUR 64 |
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Methenamine Solution |
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| MET500 | ScyTek Laboratories | 500 ml | EUR 95 |
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Methenamine Solution |
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| MET999 | ScyTek Laboratories | 1000 ml | EUR 132 |
 DTT Solution |
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| R-1104 | EpiGentek | 1 ml, 500 mM | EUR 109.45 |
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Description: fast delivery possible |
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 Tartrazine Solution |
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| TZQ125 | ScyTek Laboratories | 125 ml | EUR 64 |
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Tartrazine Solution |
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| TZQ500 | ScyTek Laboratories | 500 ml | EUR 98 |
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Tartrazine Solution |
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| TZQ999 | ScyTek Laboratories | 1000 ml | EUR 127 |
 Schiff's Solution |
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| SRF030 | ScyTek Laboratories | 30 ml | EUR 64 |
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Schiff's Solution |
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| SRF250 | ScyTek Laboratories | 250 ml | EUR 77 |
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Schiff's Solution |
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| SRF3800 | ScyTek Laboratories | 1 Gal. | EUR 345 |
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Schiff's Solution |
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| SRF500 | ScyTek Laboratories | 500 ml | EUR 103 |
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Schiff's Solution |
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| SRF999 | ScyTek Laboratories | 1000 ml | EUR 149 |
) Exosome Isolation kit (for serum & plasma -Exosome-RNA-NGS) |
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| P101NGS | 101Bio | NULL | EUR 0 |
 50ml TC Tubes, Conical, 440 units/box |
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| 04-5540150 | Gentaur Genprice | 440 units/box | EUR 71 |
 GMP IL4, 50µg |
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| 04-GMP-HU-IL4-50UG | Gentaur Genprice | 50µg | EUR 483 |
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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. |
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 rHu IL 2 , 3MIU |
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| 04-RHIL2-02F01 | Gentaur Genprice | 1 vial | EUR 249 |
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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. |
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 rHu IL 2 , 3MIU , Lot 200908F02 |
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| 04-RHIL2-08F02 | Gentaur Genprice | 1 vial | EUR 249 |
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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. |
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 PRE-GMP rHu GM-CSF, Molgramostim-Leukoma |
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| 04-RHUGM-CSF-7A10 | Gentaur Genprice | 300 µg | EUR 385 |
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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. |
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 Gentamicin Solution, Gentamicin Solution, 50 mg/mL |
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| CCM1123-010 | BBI Biotech | 10 mL | EUR 64.05 |
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 ExoQuick ULTRA EV Isolation Kit for Serum and Plasma |
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| EQULTRA-20A-1 | SBI | 20 reactions | EUR 579 |
 Exosome Component 5 Protein |
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| 20-abx260873 | Abbexa |
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 Exosome Component 7 Protein |
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| 20-abx261347 | Abbexa |
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 Exosome Component 10 Protein |
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| 20-abx261805 | Abbexa |
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 Exosome Component 4 Protein |
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| 20-abx261954 | Abbexa |
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 Exosome Component 9 Protein |
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| 20-abx262123 | Abbexa |
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 Exosome Component 3 Protein |
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| 20-abx263422 | Abbexa |
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 Exosome Component 1 Protein |
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| 20-abx263423 | Abbexa |
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 Exosome Component 8 Protein |
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| 20-abx262850 | Abbexa |
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 EXOCET Exosome Quantitation Kit |
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| EXOCET96A-1 | SBI | 96 reactions | EUR 521 |
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 FluoroCet Exosome Quantitation Kit |
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| FCET96A-1 | SBI | 96 reactions | EUR 726 |
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) Fluorescent Exosome Standard (Urine ) |
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| ESF-15 | Creative Biolabs | 100 µg | EUR 1723 |
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Description: Fluorescent exosomes are labeled with green dye, which offer a stable fluorescent labeling. |
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 Exosome-TEM-easy Kit |
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| P130 | 101Bio | NULL | EUR 0 |
 Exosome) Diabetes (Type 1) Exosome |
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| P141-DB1 | 101Bio | NULL | EUR 0 |
 Exosome) Diabetes (Type 2) Exosome |
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| P141-DB2 | 101Bio | NULL | EUR 0 |
 Leukemia - Hairy Cell Exosome |
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| P141-HC | 101Bio | NULL | EUR 0 |
 Leukemia - Multiple Myeloma Exosome |
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| P141-MM | 101Bio | NULL | EUR 0 |
 Exosome) Melanoma (skin cancer) Exosome |
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| P141-MN | 101Bio | NULL | EUR 0 |
 Non-Hodgkin lymphoma Exosome |
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| P141-NH | 101Bio | NULL | EUR 0 |
 Systemic Lupus Erythematosus Exosome |
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| P141-SL | 101Bio | NULL | EUR 0 |
 Uterine Cervical Cancer Exosome |
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| P141-UC | 101Bio | NULL | EUR 0 |
 Uterine Corpus Cancer Exosome |
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| P141-UT | 101Bio | NULL | EUR 0 |
 Normal human exosome RNA |
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| P240 | 101Bio | NULL | EUR 0 |
 Leukemia - ALL Exosome RNA |
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| P241-ALL | 101Bio | NULL | EUR 0 |
 Leukemia - AML Exosome RNA |
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| P241-AML | 101Bio | NULL | EUR 0 |
 Bladder Cancer Exosome RNA |
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| P241-BL | 101Bio | NULL | EUR 0 |
 Breast Cancer Exosome RNA |
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| P241-BR | 101Bio | NULL | EUR 0 |
 Leukemia - CLL Exosome RNA |
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| P241-CLL | 101Bio | NULL | EUR 0 |
 Leukemia - CML Exosome RNA |
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| P241-CML | 101Bio | NULL | EUR 0 |
 Colorectal Cancer Exosome RNA |
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| P241-CO | 101Bio | NULL | EUR 0 |
 Crohn's Disease Exosome RNA |
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| P241-CR | 101Bio | NULL | EUR 0 |
 Kidney Cancer Exosome RNA |
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| P241-KN | 101Bio | NULL | EUR 0 |
 Lung Cancer Exosome RNA |
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| P241-LG | 101Bio | NULL | EUR 0 |
 Liver Cancer Exosome RNA |
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| P241-LV | 101Bio | NULL | EUR 0 |
 Nasopharyngeal Cancer Exosome RNA |
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| P241-NA | 101Bio | NULL | EUR 0 |
 Prostate Cancer Exosome RNA |
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| P241-PR | 101Bio | NULL | EUR 0 |
 Rheumatoid Arthritis Exosome RNA |
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| P241-RA | 101Bio | NULL | EUR 0 |
 Thyroid Cancer Exosome RNA |
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| P241-TH | 101Bio | NULL | EUR 0 |
 HYGROMYCIN B SOLUTION |
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| 30-240-CR | CORNING | 20 mL/pk | EUR 157 |
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Description: Media Catalog; Cell Culture Reagents |
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 Ponceau S Solution |
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| 22001 | Biotium | 1L | EUR 125 |
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Description: Minimum order quantity: 1 unit of 1L |
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) HEPES SOLUTION (1M) |
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| 25-060-CI | CORNING | 100 mL/pk | EUR 104 |
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Description: Media Catalog; Cell Culture Reagents |
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 Acrylamide Solution 30% |
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| A067-1L | TOKU-E | 1 L | EUR 103 |
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Acrylamide Solution 30% |
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| A067-500Ml | TOKU-E | 500 ml | EUR 71 |
 Ringer Solution Tablets |
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| 09-1025-100 | Medicago | 100 tablets | EUR 70 |
 PROTEINASE K SOLUTION |
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| 501-PK | Viagen | 5 ml | EUR 152 |
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Description: Genomic PCR Grade Proteinase K Solution |
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 CometAssay Lysis Solution |
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| 4250-010-01 | Biotechne | 100 mL | EUR 157.8 |
 Alcian Yellow Solution |
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| ANY125 | ScyTek Laboratories | 125 ml | EUR 83 |
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Alcian Yellow Solution |
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| ANY3800 | ScyTek Laboratories | 1 Gal. | EUR 454 |
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Alcian Yellow Solution |
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| ANY500 | ScyTek Laboratories | 500 ml | EUR 123 |
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Alcian Yellow Solution |
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| ANY999 | ScyTek Laboratories | 1000 ml | EUR 178 |
 solution) Tyloxapol (10% ) solution |
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| B1159-250 | Biovision | EUR 512 | |
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Tyloxapol (10% ) solution |
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| B1159-50 | Biovision | EUR 175 | |
 Hematoxylin Counterstain Solution |
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| AR0005 | BosterBio | 12mL(for 120-180 assays) | EUR 65 |
 Ponceau S solution |
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| AR0142 | BosterBio | 12mL(10X) | EUR 60 |
 Astra Blue Solution |
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| AST125 | ScyTek Laboratories | 125 ml | EUR 104 |
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Astra Blue Solution |
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| AST500 | ScyTek Laboratories | 500 ml | EUR 183 |
 DAB Differentiating Solution |
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| DDS500 | ScyTek Laboratories | 500 ml | EUR 109 |
 Crystal Violet Solution |
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| CVA500 | ScyTek Laboratories | 500 ml | EUR 106 |
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.