Comprehensive RNA Extraction and qRT-PCR Analysis

According to the protocol, the Trizol reagent (Invitrogen) extracted the total RNA of CC cells. Next, the complementary DNA (cDNA) was obtained by transcribing RNA. RNA and cDNA quality was determined by Bioanalyzer (Agilent). The commercialized cDNA Reverse Transcription kit (Applied Biosystems, USA) was used. The level of qRT-PCR was assessed with the MicroRNA Assay and TaqMan® Universal PCR Master Mix (Applied Biosystems, USA) under the thermocycler conditions: 95°C for 5 min, and 40 cycles of 95°C for 5 s and 60°C for 31s, followed by 75°C for 40s. Besides, qRT-PCR was implemented to measure the level of mRNA and miRNA under the specific thermocycler conditions according to the agreement of the commercialization kit [21 (link)]. The primers were obtained from TAKARA (Beijing, China) and listed in Table 1.