Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway

HIC-5 expression was detected in osteosarkoma patients and osteosaroma cell lines by RT-PCR. Then RFP-SH-HIC-5 is transfected into the osteosarcoma cell line. The HIC-5 effect on cell viability, proliferation and apoptosis is assessed by MTT, Edu Kit and Flow Cytometry. Exosom is isolated from mg-63 cell supernatant by an exosomed insulation kit. Exosome-HIC-5 is confirmed by microscope electron transmission, particle size detection and RT-PCR.

Furthermore, the cells of the exosom-hic-5-5 are explored eligibility of cells, proliferation, and apoptosis. Furthermore, the C-IP test is used to identify the relationship between HIC-5 and SMAD4. TCF / LEF and the level of protein components of the WNT / β-Catenin signal are detected by the top luciferase test and Western blot. HIC-5 is regulated in osteosaroma tissue and cells. The decreased expression forced HIC-5 inhibits the proliferation of osteosarkoma cell lines, and induces the apoptosis of MG-63 and HOS. In Vivo, silencing HIC-5 sent the development of tumors. Furthermore, we isolate the exosom of the MG-63 supernatant, the excosom to conclude HIC-5 will regulate the proliferation and level of cell apoptosis of MG-63 and HOS. Furthermore, HIC-5 interacts with SMAD4 and regulates WNT / β-Catenin signals by reducing TCF / LEF activity.

Silencing HIC-5 inhibits proliferation and induces the apoptosis of cell osteosarcoma through deactivating the WNT / β-Catenin signal with the exosome path. Hypoxia is a characteristic of several serious diseases such as cardiovascular disorders and metabolism and cancer. A decrease at the oxygen level of the network induces hypoxic response in cells that seek to adapt to changing conditions. Failure to adapt to prolonged or severe hypoxia can trigger cell death. While some types of cells, such as neurons, are very susceptible to hypoxia, cancer cells utilize the hypoxia environment to undergo tumor growth, angiogenesis and metastasis. Hypoxic-induced processes trigger communication between complex cells and now there is an indication that the extracellular vesicle (EVS) plays a fundamental role in this process. The latest developments in EV isolation and the characterization methodology have raised awareness of the importance of EV purity in functional studies and cargo.

 

Exosomal limitations in the treatment of inflammatory diseases

Background: Despite great interest and many studies, there are currently no integrated standards that describe sequential manipulation with cells to get exosom for clinical use. The use of exosom has become an interesting alternative to cell therapy, because of the flexible properties of these biological vesicles. Enables scientists to manipulate their composition to produce the desired exosom to bring certain drugs, RNA and protein. This study aims to analyze the scientific literature on changes in the functional characteristics of exosom, depending on the manipulation method, which has the potential to contribute to the development of negative effects in the treatment of inflammatory genesis diseases. The choice of insulation method affects the set of protein markers disclosed, nucleic acid and receptors on microparticles.

Various surface receptors present on the exosome membrane can be engineered for the target of the lesion. Exocomals of healthy patients help reduce inflammation, normalize communication between cells and have antifibrotic, antioxidant and cytoprotective effects. Exosom can change the micro environment, but the micro environment can also change the composition of the exosomes.

Conclusion: Ecosomes obtained from sick patients carry markers of the characteristics of the appropriate disease. Such ecosomes can have pro-inflammatory properties, pro-fibrotic, cytotoxic, and oncogenic, and disrupt cellular cooperation. Until now, questions about doses, reactions to recurring giving, and the dose regime has not been fully resolved.

 Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway
Exosome-mediated Hic-5 regulates proliferation and apoptosis of osteosarcoma via Wnt/β-catenin signal pathway

Proteomic exploration based on mass spectrometry of small urinary extracellular vesicles in vasculitis associated with ANCA compared to total urine

The vasculitis associated with the ANCA (AAV) is a rare, but potentially serious self-immune disease, if not nowadays, with increased mortality and morbidity. Finding the first activity biomarkers and the prognosis is very important. The small extracellular vesicles (EVS) isolated from the urine can be considered as a non-invasive source of biomarkers. We evaluated several protocols for the isolation of urinary electric vehicles.

To eliminate contaminating non-vesicular proteins due to AAV associated proteinuria, we used the processing of proteinase K. We have studied the differences in proteomes of small patient events with AAV compared to healthy controls by the freedom of LC-MS / MS labels. In parallel, we performed a similar proteomic analysis of urine samples from identical patients. The results of the study showed significant differences and similarities in the EV proteome and urine, the latter being strongly affected by proteinuria. Using bioinformatic tools, we have explored the modified proteins differently and their related routes by focusing on AAV’s pathophysiology. Our results indicate a significant regulation of Golgi enzymes, such as MAN1A1, which may be involved in the activation of the T cell by N-glycans glycosylation and can play a key role in the pathogenesis and diagnosis of AAV.

Total RNA Isolation Kit II

FATRS--001 100 preps
EUR 369.6

Total RNA Isolation Kit II

FATRS--050 50 preps
EUR 259.2

Total RNA Isolation Kit (Plant)

P-9104 50 samples
EUR 416.7

Total RNA Isolation Kit (Plant)

PDC07-0100 100 rxns
EUR 209

Total RNA Isolation Kit (Tissue)

P-9102 50 samples
EUR 416.7

Total RNA Isolation Kit(Tissue)

PDC08-0100 100 rxns
EUR 209

Total RNA Isolation Reagent Kit

abx460023-1096tests 10 × 96 tests Ask for price

Total RNA Isolation Reagent Kit

abx460023-596tests 5 × 96 tests Ask for price

Total RNA Isolation Reagent Kit

abx460023-96tests 96 tests
EUR 275

Total RNA Isolation Reagent

PDR03-0100 100 rxns
EUR 66

EpiQuik Total RNA Isolation Fast Kit

P-9105-050 50 samples
EUR 176

EpiQuik Total RNA Isolation Fast Kit

P-9105 50 samples
EUR 163

Plant Total RNA Isolation Reagent Kit

abx460024-1096tests 10 × 96 tests Ask for price

Plant Total RNA Isolation Reagent Kit

abx460024-596tests 5 × 96 tests Ask for price

Plant Total RNA Isolation Reagent Kit

abx460024-96tests 96 tests
EUR 362.5

FastPure Cell/Tissue Total RNA Isolation Kit

RC101 50 rxn
EUR 274.8

Plant Total RNA Isolation Reagent

PDR04-0100 100 rxns
EUR 110

FastPure Universal Plant Total RNA Isolation Kit

RC411-01 50 rxns
EUR 100

FastPure Cell/Tissue Total RNA Isolation Kit V2

RC112-01 50 rxns
EUR 82.6

Total RNA Isolation Kit II (Tri-reagent + RNA spin column)

FATRS__050 1 kit
EUR 62

Total RNA Isolation Kit II (Tri-reagent + RNA spin column)

FATRS__100 1 kit
EUR 112

Total RNA Isolation Kit (Blood/Cultured Cell/Fungus)

PDC04-0100 100 rxns
EUR 220

FastPure Plant Total RNA Isolation Kit (Polysaccharides&Polyphenolics –rich)

RC401-01 50 rxns
EUR 100

FastPure Plant Total RNA Isolation Kit (Polysaccharides/Polyphenolics-Rich)

RC401 50 rxn
EUR 283.2

DNA/RNA Mini Kit (isolation of both genomic DNA and total RNA from a single sample)

FADRK_050 1 kit
EUR 90

Rna Isolation Kit - EACH

11828665001 EACH
EUR 425.95

Cartilage RNA Isolation Kit

K2031010 1 kit
EUR 487

Cell RNA Isolation(200) Kit

TBS6001 200 samples
EUR 358

Tissue RNA Isolation(200) Kit

TBS6003 200 samples
EUR 358

T-Pro Total Exosome Isolation reagent (from serum)

JO66-V002M 25ml/BT
EUR 800

T-Pro Total Exosome Isolation reagent (from serum)

JO66-V002S 1ml*5/set
EUR 200

EVery EV RNA Isolation Kit

EVery100B-1 20 preps
EUR 412

Blood RNA Isolation(50) Kit     

TBS6002 50 samples
EUR 198

RNA Broad Range Standard, 100 ng/uL

99850 1UL
EUR 63
Description: RNA from immortalized human cell line (Homo sapiens)

RNA Broad Range Standard, 100 ng/uL

99850-1 EA
EUR 63

Viral DNA/RNA Isolation Kit

AGDK-0044 Kit Ask for price
Description: The Viral DNA/RNA Isolation Kit is intended for rapid co-extraction of viral DNA and RNA from a variety of biofluid samples, such as, plasma, serum, milk and swap samples. The proprietary microspherical paramagnetic beads used in the kit have a large binding surfaces and a high affinity towards nucleic acids. Going through sample lysis/binding, washing, and elution steps, the whole process can be completed under 35 minutes, and yields highly pure nucleic acids elute.  The recovered nucleic acids can be used in a wide range of applications, such as PCR, RT-PCR, Sanger Sequencing, NGS, and gene chips.

Rapid Blood RNA Isolation Kit

BT4182 10Preps
EUR 98.36

Rapid Blood RNA Isolation Kit

BT4183 50Preps
EUR 197.03

Rapid Blood RNA Isolation Kit

BT4184 250preps
EUR 608.1

Rapid Plant RNA Isolation Kit

PT4191 50Preps
EUR 95.63

EpiMag Viral RNA Isolation Kit

P-9108-050 50 samples
EUR 152.9

EpiMag Viral RNA Isolation Kit

P-9108 50 samples
EUR 144

Easy3-Plant RNA Isolation Kit

FYG502-100P 100 Preps Ask for price

Easy3-Plant RNA Isolation Kit

FYG502-500P 500 Preps Ask for price

Rapid Bacteria RNA Isolation Kit

BS8625 50Preps
EUR 95.63

Reagent for Total Exosome Isolation (Culture Media Supplement)

EIR-02 50 ml
EUR 2485.2
Description: This product is used for isolating exosomes from culture media supplement based on chemical precipitation isolation method.

AccuBlue® Broad Range RNA Quantitation Kit (1000 assays)

31073-1 KT
EUR 341

Phenol-Free Plant RNA Isolation Kit

10760056-1 50 Prep(s)
EUR 170.53

Column-Pure Fungi RNA Isolation Kit

D510 50 Preparations
EUR 150

AccuBlue® Broad Range RNA Quantitation Kit (200 assays)

31073-T 1kit
EUR 99
Description: RNA from immortalized human cell line (H. sapiens)

AccuBlue® Broad Range RNA Quantitation Kit (200 assays)

31073-T-1 KT
EUR 99

Easy3-RNA Isolation Kit

FYG501-100P 100 Preps Ask for price

Easy3-RNA Isolation Kit

FYG501-500P 500 Preps Ask for price

T-Pro Total Exosome Isolation reagent (from cell culture media)

JO66-V001M 500ml/BT
EUR 800

T-Pro Total Exosome Isolation reagent (from cell culture media)

JO66-V001S 100ml/BT
EUR 200

EpiQuik Magbeads Quick RNA Isolation Kit

P-9106-050 50 samples
EUR 163.9

EpiQuik Magbeads Quick RNA Isolation Kit

P-9106 50 samples
EUR 151

Soil RNA Isolation Mini Kit (100prep)

FASRK-100 100 preps
EUR 224.4

Soil RNA Isolation Mini Kit (50prep)

FASRK-001 50 preps
EUR 180

EpiQuik Viral RNA Isolation Fast Kit

P-9107-050 50 samples
EUR 167.2

EpiQuik Viral RNA Isolation Fast Kit

P-9107 50 samples
EUR 158

Easy3-Total NucleicAcid Isolation Kit

FYG504-100ML 100 ml Ask for price

Easy3-Total NucleicAcid Isolation Kit

FYG504-500ML 500 ml Ask for price

RNA-easy Isolation Reagent

R701-01 100 ml
EUR 201.6

RNA-easy Isolation Reagent

R701-02 200 ml
EUR 254.4

rSYNC™ RNA Isolation Kit - Individual Packed Columns

RS004 each
EUR 3

rSYNC™ RNA Isolation Kit - Individual Packed Columns

RS050 each
EUR 83.4

rSYNC™ RNA Isolation Kit - Individual Packed Columns

RS100 each
EUR 153

rSYNC™ RNA Isolation Kit - Individual Packed Columns

RS300 each
EUR 408

miRNA Isolation Kit (Tri-reagent + RNA spin column)

FAMIK__001 1 kit
EUR 134.5

miRNA Isolation Kit (Tri-reagent + RNA spin column)

FAMIK__002 1 kit
EUR 73

Protein Marker-2, Broad Range

GWB-P0088J 500 ug Ask for price

S-adenosylmethionine ELISA kit, broad range

IK00202 96 tests
EUR 599
Description: S-adenosylmethionine ELISA kit, broad range

S-adenosylmethionine ELISA kit, broad range

IK00203 48 tests
EUR 349
Description: S-adenosylmethionine ELISA kit, broad range

S-adenosylhomocysteine ELISA kit, broad range

IK00302 96 tests
EUR 999
Description: S-adenosylhomocysteine ELISA kit, broad range

S-adenosylhomocysteine ELISA kit, broad range

IK00303 48 tests
EUR 499
Description: S-adenosylhomocysteine ELISA kit, broad range

PureXtract RNAsol RNA Isolation Solution, LS

R6103-120 2X60ml
EUR 343.2

Broad Range DNA Quantification Kit, 100 assays

NS1020-BR-100 1 each
EUR 133.93

AccuBlue® Broad Range dsDNA Quantitation Kit

31007-1 KT
EUR 295

miRNA Isolation Kit

FYG304-100P 100 Preps Ask for price

miRNA Isolation Kit

FAMIK--001 Rxns: 100
EUR 398.4
Description: Contents of the kit:
  • Rxns:100Lysis
  • Buffer-10 ml
  • Acid-Phenol-10 ml
  • RNA Column and Collection Tube-100 sets
  • Wash Buffer-10 ml
  • Release Buffer-0.5 ml

miRNA Isolation Kit

FAMIK--002 Rxns: 50
EUR 267.6

Exosome Isolation Kit

E601E001 20 tests
EUR 2500

rSYNC™ RNA Isolation Kit w/DNase Set - Individual Packed Columns

RSD004 each
EUR 5

rSYNC™ RNA Isolation Kit w/DNase Set - Individual Packed Columns

RSD050 each
EUR 103.4

rSYNC™ RNA Isolation Kit w/DNase Set - Individual Packed Columns

RSD100 each
EUR 193

rSYNC™ RNA Isolation Kit w/DNase Set - Individual Packed Columns

RSD300 each
EUR 518

MicroRNA Isolation Kit

KS341025 1 kit
EUR 284

Endosome Isolation Kit

P528 - Ask for price

Microsome Isolation Kit

K249-50 each
EUR 633.6

Protoplast Isolation Kit

30210002-1 20 Prep(s)
EUR 199.16

Protoplast Isolation Kit

30210002-2 50 Prep(s)
EUR 438.32

Chloroplast Isolation Kit

30210000-1 20 Prep(s)
EUR 273.97

Magnetic Pathogens Nucleic Acid (DNA/RNA)Isolation Kit, 50 reactions/pk

208601 8 pcs/cs
EUR 131.12
Description: Magnetic Pathogens Nucleic Acid (DNA/RNA)Isolation Kit, 50 reactions/pk

CELLDATA RNAstorm™ Fresh Cell and Tissue RNA Isolation Kit, 50 preps

CD504 1
EUR 270
Description: DNAse I from bovine pancreas (cow; Bos taurus)

CELLDATA RNAstorm™ Fresh Cell and Tissue RNA Isolation Kit, 50 preps

CD504-1 KT
EUR 270

Pharmalyte broad range pH 8-10.5 25mL - EACH

17045501 EACH
EUR 847.8

Pharmalyte broad range pH 3-10 25mL - EACH

17045601 EACH
EUR 708.75

Mitochondria Isolation Kit

AR0156 1 kit
EUR 127.2

Preadipocyte Isolation Kit

K583-5 each
EUR 639.6

Urine DNA Isolation Kit

K5011150 1 kit
EUR 261

Stool DNA Isolation Kit

K5011250 1 kit
EUR 261

Serum DNA Isolation Kit

K5018100 1 kit
EUR 391

Saliva DNA Isolation Kit

AG0226 Kit Ask for price
Description: Saliva DNA Isolation Kit provides a fast and simple spin column procedure for isolating high quality DNA from saliva samples collected and preserved using AcceGen's Saliva DNA Collection and Preservation Devices, as well as fresh saliva samples.Saliva DNA purified using AcceGen's kit is of the excellent quality, and is compatible with a number of downstream research applications including PCR, qPCR, Southern Blot analysis, sequencing and microarray analysis.

Saliva DNA Isolation Kit

K5011050 1 kit
EUR 261

Genomic DNA Isolation Kit

55R-1362 50 assays
EUR 273
Description: Genomic DNA Isolation Kit for use in the research laboratory

Genomic DNA Isolation Kit

GWB-AXR187 50 assays Ask for price

Genomic DNA Isolation Kit

K2118-50 50 assays
EUR 389
Description: Isolates highly pure Genomic DNA within 90 min.

Genomic DNA Isolation Kit

K281-50 each
EUR 326.4

Genomic DNA Isolation Kit

abx460025-1096tests 10 × 96 tests Ask for price

Genomic DNA Isolation Kit

abx460025-596tests 5 × 96 tests Ask for price

Genomic DNA Isolation Kit

abx460025-96tests 96 tests
EUR 437.5

Yeast Nuclei Isolation Kit

K289-50 each
EUR 470.4

ExoPure? Isolation Kit   (Urine)

K1240-10 each
EUR 836.4

ExoPure? Isolation Kit   (Urine)

K1240-2 each
EUR 483.6

Lipid Droplet Isolation Kit

MET-5011 50 preps
EUR 345

Prosi Prestained Protein Marker, Broad Range(11-170KDa)

P8500-040 200ulX2
EUR 271.2

Apoptotic Cell Isolation Kit

55R-1349 30 units
EUR 513
Description: Apoptotic Cell Isolation Kit for use in the research laboratory

miRNA ISOLATION KIT-TISSUE

IB47370 4 PREP KIT
EUR 30.42

miRNA ISOLATION KIT-TISSUE

IB47371 100 PREP KIT
EUR 579.66

Apoptotic Cell Isolation Kit

GWB-AXR172 30isolations Ask for price

Apoptotic Cell Isolation Kit

K258-30 each
EUR 561.6

Exosome Isolation kit (for serum & plasma -Exosome-RNA-NGS)

P101NGS - Ask for price

DNA Ladder One(Broad Range)(Ready To Use)

08362-85 500UL
EUR 105

Cell Mitochondria Isolation Kit

abx090630-50100tests 50-100 tests
EUR 309.6

Mitochondrial DNA Isolation Kit

55R-1361 50 assays
EUR 660
Description: Mitochondrial DNA Isolation Kit for use in the research laboratory

Mitochondrial DNA Isolation Kit

GWB-AXR186 50 assays Ask for price

Cell Mitochondria Isolation Kit

ETF3601 100 assays
EUR 135

Mitochondrial DNA Isolation Kit

K2117-1 1 kit
EUR 686
Description: Isolates mt. DNA, no genomic DNA contamination

Mitochondrial DNA Isolation Kit

K2117-50 50 assays
EUR 718.8

Mitochondrial DNA Isolation Kit

K280-50 each
EUR 705.6

Cell Mitochondria Isolation Kit

abx090630-100l 100 µl
EUR 187.5

Cell Mitochondria Isolation Kit

abx090630-1ml 1 ml Ask for price

Cell Mitochondria Isolation Kit

abx090630-200l 200 µl Ask for price

Exo-Urine EV Isolation Kit

EXOU100A-1 2 x 5 reactions
EUR 490

FitAmp Gel DNA Isolation Kit

P-1007 100 Samples
EUR 324.48

Pharmalyte broad range pH 2.5-5 25mL - EACH

17045101 EACH
EUR 847.8

Pharmalyte broad range pH 4-6.5 25mL - EACH

17045201 EACH
EUR 847.8

Meaning: The current study explores for the first time the changes in proteomes of small extracellular vesicles and urine of patients with vasculitis associated with former kidney relative to healthy controls by LC-MS / MS label. The isolation of vesicles from proteinuric urine samples has been modified to minimize plasma protein contamination and reduce co-insulation of extramuminal proteins. Differentially modified proteins and related routes with a role in AAV pathophysiology have been described and discussed. The results could be useful for the search for potential biomarkers in renal vasculitis associated with ANCa.