miRNA-126-3p regulates the proliferation and cycle change of human monocytic leukemia THP-1 cells
10.3760/cma.j.cn115356-20230519-00117
- VernacularTitle:miRNA-126-3p调控人单核细胞白血病THP-1细胞的增殖和周期变化
- Author:
Gang TI
1
;
Lili LI
;
Feng LI
Author Information
1. 山西医学科学院 山西白求恩医院 同济山西医院病案室，太原 030032
- Keywords:
Leukemia, monocytic, acute;
miRNA-126-3p;
RGS3 protein, human;
Cell proliferation;
Cell cycle
- From:
Journal of Leukemia & Lymphoma
2023;32(12):740-744
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of miRNA-126-3p (miR-126-3p) on proliferation and cycle of human monocytic leukemia THP-1 cells.Methods:Real-time fluorescence quantitative polymerase chain reaction (PCR) was used to verify the expression of miR-126-3p in THP-1 cells and monocytes isolated from the peripheral blood of healthy check-ups collected from May 2022 to January 2023 in Shanxi Cancer Hospital. The miR-126-3p overexpression vector was constructed and transfected inyo THP-1 cells (miR-126-3p overexpression group), and THP-1 cells transfected with the empty vector were used as the control. The proliferative ability of cells in each group was detected by CCK-8 assay, and the cell cycle was detected by flow cytometry. TargetScan software was applied to predict the miR-126-3p target gene as RGS3, which was verified by dual luciferase reporter gene assay. The protein expression level of RGS3 in THP-1 cells of the miR-126-3p overexpression group was detected by Western blotting.Results:The results of PCR showed that the expression level of miR-126-3p in THP-1 cells was low compared with that in monocytes isolated from peripheral blood of healthy individuals ( P < 0.05). Compared with the control group, THP-1 cells in the miR-126-3p overexpression group had low proliferative capacity after 48 h and 72 h of culture (both P < 0.05), and the cells were blocked in G 1 phase [proportion of cells in G 1 phase: (58.2±2.8)% vs. (44.1±2.4)%, P < 0.05]. The results of dual luciferase reporter gene assay showed that miR-126-3p bound to the 3'UTR of RGS3. Western blotting assay showed that the RGS3 protein expression level in THP-1 cells of the miR-126-3p overexpression group was lower than that in the control group. Conclusions:miR-126-3p may inhibit proliferation of human monocytic leukemia THP-1 cells and promote G 1-phase blockade by inhibiting the expression of RGS3.
