Musashi RNA-binding protein 2 promotes hepatocellular carcinoma growth through the Wnt/β-catenin
10.3760/cma.j.cn113884-20220422-00183
- VernacularTitle:Musashi RNA结合蛋白2通过Wnt/β-catenin信号通路对肝癌细胞增殖的影响
- Author:
Yanjun LI
1
;
Haichao ZHAO
Author Information
1. 山西白求恩医院肝胆外科，太原 030032
- Keywords:
Carcinoma, hepatocellular;
Cell proliferation;
Wnt signaling pathway;
Beta-catenin
- From:
Chinese Journal of Hepatobiliary Surgery
2022;28(10):766-771
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the mechanism that how Musashi RNA-binding protein 2 (MSI2) regulates HCC growth.Methods:Short hairpin RNA (shRNA) was transfected to inhibit MSI2 expression, and cells were divided into transfection control plasmid (sh-Ctrl) group and sh-MSI2 group. In the MSI2 overexpression experiment, cells were divided into control group (Vector group, transfected with blank plasmid Vector) and overexpression group (MSI2 group, transfected with MSI2 recombinant plasmid). Cell proliferation was detected by CCK-8 and plate cloning assays. Western blotting was used to detect the expressions of β-catenin, transcription factor 7 (TCF7) and lymphoid enhancer factor 1 (LEF1) after MSI2 intervention. In the Rescue recovery experiment, cells were divided into MSI2+ sh-Ctrl group (transfected with MSI2 recombinant plasmid and sh-Ctrl plasmid at the same time) and MSI2+ sh-β-catenin group (transfected with MSI2 recombinant plasmid and sh-β-catenin plasmid at the same time). On the basis of overexpression of MSI2, β-catenin was knocked down to detect the proliferation ability of HCC cells.Results:The proliferation rates of HepG2 and MHCC97H cells in the sh-MSI2 group were lower than those in the sh-Ctrl group, and the difference was statistically significant ( P<0.05). The proliferation rates of SMMC-7721 cells and MHCC97L cells in the MSI2 group were higher than those in the Vector group, and the difference was statistically significant ( P<0.05). The results of the clone formation experiment showed that compared with the sh-Ctrl group, the number of HepG2 cell clones in the sh-MSI2 group [(129.7±6.5) vs. (286.0±12.8)] and the number of MHCC97H cell clones [(134.0±6.7) vs. (248.0±14.1)] were reduced, and the differences were statistically significant ( P<0.05); compared with the Vector group, the number of SMMC-7721 cell clones [(242.0±5.6) vs. (135.3±8.7)] and MHCC97L cell clones [(308.0±9.0) vs. (149.7±5.9)] in the MIS2 group were increased, and the difference was statistically significant ( P<0.05). The mRNA and protein expression of β-catenin, TCF7 and LEF1 in HepG2 and MHCC97H cells in the sh-MSI2 group were lower than those in the sh-Ctrl group, and the differences between groups were statistically significant ( P<0.05). In contrast, compared with the Vector group, the mRNA and protein expression levels of β-catenin, TCF7 and LEF1 in SMMC-7721 and MHCC97L cells in the MSI2 group were all increased, and the differences were statistically significant ( P<0.05). Compared with the MSI2+ sh-Ctrl group, the cell proliferation ability of the MSI2+ sh-β-catenin group was decreased, and the difference was statistically significant ( P<0.05). Plate cloning experiments showed that the number of cell clones in the MSI2+ sh-β-catenin group was less than that in the MSI2+ sh-Ctrl group [(138.3±7.0) vs. (246.3±8.0), P=0.028]. Conclusion:MSI2 promotes HCC cell proliferation through Wnt/β-catenin signaling pathway, leading to tumor progression.
