|
| Status |
Public on May 03, 2023 |
| Title |
mi2_8-P |
| Sample type |
RNA |
| |
|
| Source name |
colorectal tumor tissues of CPC-Apc mice as “Tumor”, replicate 1
|
| Organism |
Mus musculus |
| Characteristics |
tissue: colorectal tumor
genotype/variation: CPC-Apc mice
|
| Treatment protocol |
We performed treatment of the combination of miRNAs to mouse model of spontaneous colorectal cancer, using CDX2P-NLS Cre;Apc+/loxP (CPC-Apc) mice. CPC-Apc mice were treated twenty-four times with formulated miRNAs (miR-200c, miR-302 (-a,-b,-c,-d), and miR-369 (-3p, -5p)) or negative control miRNA, three times a week from 8 weeks of age. MiRNAs were injected via tail vein using super carbonate apatite (sCA) as a drug delivery system. Briefly, 350 μL of 1 M CaCl2 was mixed with 25 μg of each miRNA or with a commensurate amount of NC miRNA in 87.5 mL of serum-free bicarbonate (44 mM)-buffered inorganic solution per mouse, and subsequently the mixture was incubated at 37°C for 30 min. The solution was centrifuged at 12,000 rpm for 3 min, and the pellet was dissolved with saline containing 0.5% albumin. The products were sonicated (38 kHz, 80 W) in a water bath for 10 min to generate sCA+0.5% albumin, which were intravenously injected within 10 min. At the time of sacrifice, colorectal tissue was collected and divided into normal tissues and tumor tissues for both groups of mice.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated with TRIzol Reagent (Invitrogen) for mouse samples following the manufacturer’s protocol. RNA quality was assessed with NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies).
|
| Label |
Cy3
|
| Label protocol |
31 ng of total RNA was dephosphorylated with calf intestine alkaline phosphatase, denatured with dimethyl sulfoxide, and labeled with pCp-Cy3 using T4 RNA ligase using the miRNA Labeling Reagent and Hybridization Kit.
|
| |
|
| Hybridization protocol |
Refer to Agilent miRNA hybridization protocol. Probes were hybridized at 55°C for 20 hours with rotation. Then the slides were washed by Gene expression wash buffer 1 at room temperature for 5 minutes and by Gene expression wash buffer 2 at 37°C for 5 minutes.
|
| Scan protocol |
After hybridization and washing, the slides were scanned using an Agilent scanner (G2505C). Images were extracted using Agilent Feature Extraction software (ver. 10.7.3.1) and Agilent GeneSpring GX software (ver. 13).
|
| Description |
MR-P
|
| Data processing |
Raw data were analyzed using Agilent GeneSpring GX software (ver.13).
|
| |
|
| Submission date |
Dec 27, 2016 |
| Last update date |
May 03, 2023 |
| Contact name |
Daisuke Okuzaki |
| E-mail(s) |
dokuzaki@biken.osaka-u.ac.jp
|
| Phone |
+81-6-6879-4935
|
| Organization name |
Osaka univ.
|
| Department |
Immunology Frontier Research Center
|
| Lab |
Human Immunology （Single Cell Genomics)
|
| Street address |
Yamadaoka 3-1
|
| City |
Suita |
| State/province |
Osaka |
| ZIP/Postal code |
565-0871 |
| Country |
Japan |
| |
|
| Platform ID |
GPL22861 |
| Series (2) |
| GSE92953 |
Inhibitory effect of microRNA on carcinogenesis of colorectal cancer using CDX2P-NLS Cre;Apc+/loxP (CPC-Apc) mice [miRNA] |
| GSE92974 |
Inhibitory effect of microRNA on carcinogenesis of colorectal cancer using CDX2P-NLS Cre;Apc+/loxP (CPC-Apc) mice |
|
