Introduction
Material and methods
BEND cell culture
Experimental design
RT-qPCR for miRNAs and mRNAs
Forward primer (5’-3’) | Reverse primer (5’-3’) | Product length (bp) | Annealing temperature (°C) | Gene bank accession | |
---|---|---|---|---|---|
PTGS2 | CTG AGT ACT TTT GAC TGT GGG AG | CTC TTC CTC CTG TGC CTG AT | 359 | 60 | NM_174445 |
PLA2G4A | AAA TGT CAG CCA CAA CCC TC | ATG GAG GGT GAA AAG CG | 229 | 56 | NM_001075864.1 |
PR | GAG AGCT CAT CAA GGC AAT TGG | CAC CAT CCC TGC CAA TAT CTTG | 227 | 60 | NM_001205356.1 |
ESR1 | AGG GAA GCT CCT ATT TGC TCC | CGG TGG ATG TGG TCC TTC TCT | 234 | 58 | AY538775 |
SDHA | GGG AGG ACT TCA AGG AGA GG | CTC CTC AGT AGG AGC GGA TG | 219 | 60 | DQ386895.1 |
SUZ12 | TTC GTT GGA CAG GAG AGA CC | GTG CAC CAA GGG CAA TGT AG | 286 | 60 | NM_001205587.1 |
ACTB | CGG TGC CCA TCT ATG AGG | GAT GGT GAT GAC CTG CCC | 266 | 58 | AY141970 |
GAPDH | CCC AGA AGA CTG TGG ATG G | AGT CGC AGG AGA CAA CCT G | 306 | 32 | U85042 |
Primer sequence (5’-3’) | ||
---|---|---|
bta-miR-106a | RT6-miRNA | TGT CAG GCA ACC GTA TTC ACC GTG AGT GGT TAC CTG |
miRNA-rev | CGT CAG ATG TCC GAG TAG AGG GGG AAC GGC GAA AAG TGC TTA CAG TG | |
bta-miR-99a-5p | RT6-miRNA | TGT CAG GCA ACC GTA TTC ACC GTG AGT GGT ACA AGA |
miRNA-rev | CGT CAG ATG TCC GAG TAG AGG GGG AAC GGC G AAC CCG TAG ATC CGA TCT |
Statistical and bioinformatics analysis
Results and discussion
Regulation of PTGS2 and PLA2G4A
IFNT upregulates PR and PDBU downregulates estrogen and progesterone receptors
Expression of miR-106a is regulated by IFNT
IRF-1 and IRF-6 are found in the promoter regions of regulated genes
IRF-1 | IRF-2 | IRF-6 | IRF-9 | |
---|---|---|---|---|
Protein coding genes promoters | 2603 | 83 | 40315 | 5 |
miRNA coding genes promoters | 54 | 2 | 1135 | 0 |
IRF | Number of binding sites | Gene |
---|---|---|
IRF-1 | 1 |
PLA2G4A
|
IRF-6 | 1 |
PR
|
IRF-6 | 1 |
PTGS2
|
IRF-6 | 1 |
ESR1
|
IRF-6 | 1 |
PLA2G4A
|
IRF-6 | 3 |
MIR106A
|