RNAiso plus (Takara, Japan) was used for total RNA isolation according to manufacturer’s instructions. A HyperScript III miRNA 1st Strand cDNA Synthesis Kit (NovalBio, China), a CR0232 lncRNA First Strand cDNA Synthesis Kit (Cellregen,China), and Hifair® III 1st Strand cDNA Synthesis SuperMix for qPCR (Yeasen, China) were used for miRNA reverse transcription, lncRNA reverse transcription, and mRNA reverse transcription, respectively. Quantitative PCR was performed to analyze cDNA using an AB 7500 Real-Time PCR System with SYBR Green (Takara). GAPDH and U6 were used as the internal controls. The relative expression level of RNAs was determined using the 2−ΔΔCt method. The sequences of the primers used are presented in Table 1 .
The Sequences of the Primers in This Study
| Primer | Sequences |
|---|---|
| miR-3940-3p | Forward: 5′-CTCAAGGACCACCGCATC-3′ |
| Reverse: 5′-ATCTGCAAGGGACAGCACAG-3′ | |
| BBOX1-AS1 | Forward: 5ʹ-TGTGTGTTTCCTGAGGCCTC-3’ |
| Reverse: 5ʹ-CGCCTCTCTTGGAACACCTT-3’ | |
| KPNA2 | Forward: 5′-CTGGGACATCAGAACAAACCAAG-3′ |
| Reverse: 5′-ACACTGAGCCATCACCTGCAAT-3′ | |
| GAPDH | Forward: 5ʹ-CTCCTCCTGTTCGACAGTCAGC-3’ |
| Reverse: 5ʹ-CCCAATACGACCAAATCCGTT-3’ | |
| U6 | Forward: 5ʹ-CTCGCTTCGGCAGCACA-3’ |
| Reverse: 5ʹ-AACGCTTCACGAATTTGCGT-3’ |