Quantitative real-time polymerase chain reaction (qRT-PCR)

The total RNA was obtained from cells and clinical samples using the RNAiso Plus (TaKaRa, Otsu, Shiga, Japan) and Trizol LS Reagent (TaKaRa), respectively. Then the reliability of the obtained RNA was verified by formaldehyde denaturation electrophoresis. Reverse transcription was performed using the PrimeScript™ RT Reagent Kit (TaKaRa) based on the instructions. The quantification of mRNA expressions was performed by standard real-time-PCR protocol with SYBR Premix Ex Taq (TaKaRa). Glycer aldehyde-3-phosphate dehydrogenase (GAPDH) served as a reference gene and the primers are exhibited in Table 1.

Note: qRT-PCR, quantitative real-time polymerase chain reaction; lncRNA PVT1, long non-coding RNA plasmacytoma variant translocation 1; MAPK, mitogen-activated protein kinase; NF-κB, Nuclear factor-kappa B; MMP, matrix metalloproteinase; TIMP-1, tissue inhibitor of metalloproteinase-1; CRP, C reactive protein; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; F, forward; R, reversed.