Background
Methods
Human tissue samples
Identification of HPV type
RNA extraction and qRT-PCR
Cell line culture
RNA-fluorescence in situ hybridization (FISH) assay
RNaseR resistance assay
Northern blot assay
Western blot assay
Gene knockdown and overexpression
Cell viability, migration, invasion and apoptosis assays
Extracellular acidification rate (ECAR)
Inhibition of glycolysis in cells
Biotin-labeled RNA pull-down assay and mass spectrometry analysis
RNA immunoprecipitation (RIP) assay
Determination of mRNA half-life
Design and synthesis of inhibitory peptides
Biotin peptide pull-down assay
Nude mice xenograft experiments
MicroPET/CT scans in nude mice
Digestion of subcutaneous tumors
Detection of LA and PA
Zebrafish embryo model
Immunohistochemical staining
Statistical analysis
Results
CircCDKN2B-AS1 is upregulated in cervical cancer and precancerous tissues
circCDKN2B-AS1 expression | |||
---|---|---|---|
High | Low | P value | |
Age (years) | 0.203 | ||
< 35 | 6 | 4 | |
≥ 35 | 51 | 14 | |
FIGO stage | 0.022 | ||
IB | 34 | 16 | |
IIA | 23 | 2 | |
Tumor size (cm) | 0.029 | ||
< 4 | 35 | 16 | |
≥ 4 cm | 22 | 2 | |
Deep stromal invasion | |||
< 66% | 20 | 13 | 0.006 |
≥ 66% | 37 | 5 | |
Lymph nodes Metastasis | 0.044 | ||
negative | 46 | 18 | |
positive | 11 | 0 |