Sybr green

We used TRIzol reagent (TaKaRa, Otsu, Shiga, Japan) to extract RNA from tissues and cell lines, and then used NanoDrop to determine the concentration of the extract to quantify RNA. Next, we used miRNA first-strand cDNA synthesis kit (Applied Biosystems, Foster City, CA) to reverse transcribe miRNA. According to the manufacturer’s instructions, we performed qRT-PCR on the 7900 real-time PCR system (Applied Biosystems, Foster City, CA) and used SYBR Green (GenePharma, Shanghai, China) to detect the miR-4521 expression in the tissues and cell lines. U6 was selected as the endogenous control and the relative value of its expression level was calculated by the 2^−ΔΔCt method.

As per the manufacturer's guide, whole RNA from SW620/R and HCT116/R cells was acquired with the help of a Trizol reagent (Ambion). cDNA was prepared with the help of a reverse transcription system kit (Genepharma). qRT‐PCR was carried out using SYBR Green (Genepharma), and the GAPDH was kept as the endogenous reference on an Agilent Mx3005P fluorescence quantitative PCR instrument to assess the mRNA's relative expression. The tRF's relative expression was measured by the 2^−ΔΔCt method. Table S2 presents the details of the primer sequences.