Effect of microRNA-146a-5p on invasion, migration and expression of ZBTB2 of hepatoma cells
WU Tao
CHEN Enhong
JIN Cheng
LIU Minfeng
Abstract：Objective To investigate the effect of microRNA-146a-5p (miR-146a-5p) on invasion, migration and expression of zinc finger and BTB domain containing 2 (ZBTB2) of hepatoma cells. Methods The miR-146a-5p levels of normal human hepato-cyte LO2 and hepatocarcinoma SMMC-7721 cells were detected by real-time quantitative PCR (QPCR). SMMC-7721 cells were trans-fected in vitro. According to the different transfection materials, SMMC-7721 cells were divided into enhanced transfection group ( transfection with miR-146a-5p mimics), inhibited transfection group (transfection with miR-146a-5p inhibitor) and blank transfection group (transfection with only liposome). The miR-146a-5p level, proliferation activity, number of transmembrane cells and ZBTB2 lev-el were detected by QPCR, MTT colorimetry, Transwell cell migration and invasion test and Western blotting. The specific seed se-quence containing 3’ untranslated region (3’UTR) of ZBTB2 gene was cloned into Luciferase Report plasmid psiCHECK-2 and the targeting relationship between miR-146a-5p and ZBTB2 was verified by double Luciferase Report gene system. Results The miR-146a-5p level in SMMC-7721 cells was 0. 156± 0. 047, lower than 1. 157± 0. 413 in LO2 cells ( P＜0. 05). Compared with 1. 241 ± 0. 375 in blank transfection group, miR-146a-5p level in enhanced transfection group increased to 8. 159±1. 274, while that in inhibi-ted transfection group decreased to 0. 219±0. 036; compared with blank transfection group, the proliferation activity in the enhanced transfection group decreased at 48 and 72 h after transfection, while those in inhibited transfection group increased; compared with 22. 069±3. 152 in blank transfection group in the cell invasion experiment, the number of transmembrane cells in enhanced transfection group decreased to 11. 847±2. 031, while that in inhibited transfection group increased to 69. 540± 4. 257; compared with 47. 338± 5. 067 in blank transfection group in the cell migration experiment, the number of transmembrane cells in enhanced transfection group decreased to 29. 640±3. 652, while that in inhibited transfection group increased to 136. 426±18. 469; compared with 0. 564±0. 089 in blank transfection group, the ZBTB2 level in enhanced transfection group decreased to 0. 168±0. 023, while that in inhibited transfec-tion group increased to 0. 826±0. 157; the difference among groups was statistically significant (P＜0. 05). After treatment with miR-146a-5p mimics, the relative fluorescence activity of cells transfected with wild-type ZBTB2 3’UTR plasmid was lower than that of the mutant plasmid (P＜0. 05). Conclusion MiR-146a-5p is low expression in hepatoma cells and plays an anti-tumor role. This miRNA may target ZBTB2 to inhibit the migration and invasion of hepatoma cells, which has a certain application prospect in the targeted treat-ment of hepatoma.
Keywords：Hepatoma cellsMicroRNA-146a-5pInvasion and migrationZinc finger and BTB domain containing 2
Publication Date：2019-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 984-988 )