|
| Status |
Public on Mar 04, 2020 |
| Title |
HSC-4 EV |
| Sample type |
RNA |
| |
|
| Source name |
Human oral cancer cell line HSC-4 EV
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HSC-4
cell type: oral squamous cell carcinoma
genotype/variation: EV lentivirus (control)
|
| Treatment protocol |
HSC-3 and HSC-4 were infected with lentivral EV or overexpressed ORAOV1-B, stably cell lines were generated by puromycin selection.
|
| Growth protocol |
Culture oral squamous cell carcinoma cell line (HSC-3 and HSC-4) in Dulbecco’s modified Eagle’s medium (DMEM) supplied with 10% fetal bovine serum and penicilin /streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Trizol isolation system (Invitrogen) according to manufacturer’s instructions. Total RNA quality was then determined. Reverse transcription was performed using High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems) to get cDNA.
|
| Label |
Cy3
|
| Label protocol |
cDNA labeled with a fluorescent dye (Cy3-dCTP) was produced by Eberwine’s linear RNA amplification method and subsequent enzymatic reaction upon using CapitalBio cRNA Amplification and Labeling Kit (CapitalBio)
|
| |
|
| Hybridization protocol |
Labeled controls and test samples with Cy3-dCTP were dissolved in 80 L hybridization solution containing 3×SSC, 0.2% SDS, 5×Denhardt’s solution and 25% formamide. DNA in hybridization solution was denatured at 95℃ for 3 min prior to loading onto a microarray. Arrays were hybridized was performed in a Agilent Hybridization Oven overnight at a rotation speed of 20 rpm at 42℃ and washed with two consecutive solutions (0.2% SDS, 2× SSC at 42℃ for 5 min, and 0.2× SSC for 5 min at room temperature)
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2565CA)
|
| Description |
Gene expression in EV set of OSCC cells
C V
|
| Data processing |
The array data were analyzed for data summarization, normalization and quality control by using the GeneSpring software V12 (Agilent)
|
| |
|
| Submission date |
Feb 20, 2020 |
| Last update date |
Mar 06, 2020 |
| Contact name |
Qianming Chen |
| Organization name |
Sichuan University
|
| Department |
oral medicine
|
| Street address |
Renmin Nan Street Section 3 NO.14
|
| City |
Chengdu |
| State/province |
Sichuan |
| ZIP/Postal code |
610041 |
| Country |
China |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE145599 |
The function and mechanism about how ORAOV1-B potentiates the OSCC metastasis. |
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