|
| Status |
Public on Mar 10, 2023 |
| Title |
HCB514_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Cervical cancer cell line HCB-514
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: HCB-514
origin: Endocervical adenocarcinoma; HPV-16 positive
Sex: Female
age: 30 years
|
| Growth protocol |
HaCaT: were cultured in Dulbecco’s modified Eagle’s media (DMEM; Sigma) supplemented with 10% FBS (Thermo Fisher Scientific) at a humidified incubator with 5% CO2 at 37°C. HCB-514: were cultured in DMEM to one part Ham’s F12 media (F12; Sigma). The media was then supplemented with 1% P/S, 0.4 µg/mL hydrocortisone, 2.5 µg/mL insulin, 13 ng/mL liothyronine, 5 µg/mL transferrin, 2.5 µg/mL insulin and 0.1 µg/mL cholera enterotoxin and 5% FBS (all supplements were purchased from Sigma). Cells were maintained in a T25 flask at 37 °C, 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from cervical cancer cells using RecoverAll Total Nucleic Acid Isolation Kit (Thermo Fisher Scientific, Waltham, MA, USA) following the manufacturer's instructions. After, the purity and concentration of total RNA were evaluated by NanoDrop Spectrophotometer v3.7 (Thermo Fisher Scientific, Waltham, MA, USA).
|
| Label |
not provided
|
| Label protocol |
not provided
|
| |
|
| Hybridization protocol |
3 µL of sample containing 100 ng of total RNA were used, a reporter and a capture probe were added and hybridized in the respective genes of interest, according to the manufacturer's instructions (NanoString Technologies).
|
| Scan protocol |
the cartridge containing immobilized and aligned reporter complexes were transferred to nCounter® Digital Analyzer (NanoString Technologies), with 280 fields of view. These results were tabulated in RCC (Reporter Code Count) format.
|
| Data processing |
Raw data were collected and pre-processed by nSolver™ Analysis Software v4.0 (NanoString Technologies), and further normalization and differential expression analysis were performed in the R statistical environment (R-project v3.6.3; The R Foundation, Viena, Austria). The data were normalized using the housekeeping method by the NanoStringNorm package (Bioconductor). Normalized data were log2-transformed and used as input for the differential expression analysis.
|
| |
|
| Submission date |
Feb 01, 2023 |
| Last update date |
Mar 10, 2023 |
| Contact name |
Rhafaela Lima Causin |
| E-mail(s) |
rhafaela-lima@hotmail.com
|
| Organization name |
Barretos Cancer Hospital
|
| Department |
Molecular Oncology Research Center
|
| Lab |
CPOM
|
| Street address |
Antenor Duarte Villela, 1331
|
| City |
Barretos |
| State/province |
SP |
| ZIP/Postal code |
14784-400 |
| Country |
Brazil |
| |
|
| Platform ID |
GPL33084 |
| Series (1) |
| GSE224301 |
MicroRNA‑130a‑3p inhibition suppresses cervical cancer cell progression |
|
