Quantitative Real-Time PCR

Total RNA from cells was extracted with TRIeasy Total RNA Extraction Reagent TRIeasy (YEASEN, Shanghai, China) following the manufacturer’s recommendation. Total RNA from paraffin section was extracted using RNAprep Pure FFPE Kit (TIANGEN, Beijing, China). cDNA was synthesized by using the M-MLV (Vazyme Biotech, China) following standard protocols. Quantitative real-time PCR was performed by using the Applied Biosystems StepOnePlus system. mRNA levels were normalized to GAPDH. EzOmics SYBR qPCR mix and miR-873-5p qPCR kit were purchased from Biomics. miR-873-5p level was normalized to U6 snRNA. The primer sequences were mentioned in Table S2. The relative gene expression levels were calculated using the 2^−ΔΔct method.