1. 转染IGF2BP3基因siRNA和真核表达质粒的 神经母细胞瘤细胞侵袭凋亡变化观察.
- Author
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朱凯, 王志茹, 高婷婷, and 吕志宝
- Abstract
Objective To observe the changes in invasion and apoptosis of neuroblastoma (NB) cells transfected with insulin-like growth factor 2 mRNA binding protein 3 (IGF2BP3) gene siRNA and eukaryotic expression plasmid, in order to investigate the effects of IGF2BP3 gene on invasion and apoptosis of NB cells. Methods NB cell lines IMR-32, SK-N-BE (2), BE (2)-C, and SH-SY-5Y were selected. The protein and mRNA expression levels of IGF2BP3 were detected by fluorescence quantitative PCR (qRT-qPCR) and Western blotting (WB), and Transwell assay was used to detect the invasive capacity of NB cell lines in vitro. SK-N-BE (2) cells with the highest expression level of IGF2BP3 and IMR-32 cells with the lowest expression level of IGF2BP3 were selected as the follow-up experiment cells. SK-N-BE (2) cells were transfected with siRNA targeting IGF2BP3 gene (SK-siIGF2BP3 group), and meanwhile, we set up a control group (SK- Control group, only liposomes) and a siRNA control group (SK-siNC group, transfected with the negative control sequence). IMR-32 cells were transfected with IGF2BP3 eukaryotic expression plasmid (IMR-IGF2BP3 group), and meanwhile, we set up a control group (IMR-Control group, only liposomes) and a empty vector control group (IMR-Vector group, transfected with the empty vector). After successful transfection, IGF2BP3 mRNA and protein of each group were detected by qRT-PCR, Western blotting, and immunofluorescence staining. The cell invasion ability of each group was observed by Transwell assay, and the apoptosis rate of NB cells in each group was measured by flow cytometry. The invasion related proteins E-cadherin, N-cadherin and vimentin were detected by Western blotting. Results Compared with the SK-Control group and SK-siNC group, the relative expression levels of IGF2BP3 mRNA and protein decreased in the SK-siIGF2BP3 group, the cells penetrating the Transwell membrane decreased, and the apoptosis rate increased (all P<0. 05). Compared with the IMR-Control group and IMR-Vector group, the relative mRNA and protein expression levels of IGF2BP3 increased, the cells penetrating the Transwell membrane increased, and the apoptosis rate decreased in the IMR-IGF2BP3 group (all P<0. 05). In the SK-N-BE (2) cells, compared with the SK-siNC group and SK-Control group, the relative expression levels of N-cadherin and Vimentin proteins decreased, while the relative expression level of E-cadherin protein increased in the SK-siIGF2BP3 group (all P<0. 05). In the IMR-32 cells, compared with the IMR-Vector group and IMR-Control group, the relative expression levels of N-cadherin and Vimentin proteins increased, while the relative expression level of E-cadherin protein decreased in the IMR-IGF2BP3 group (all P<0. 05). Conclusion The invasive ability of NB cells transfected with siRNA of IGF2BP3 gene decreased and the apoptosis rate increased, while the invasive ability of NB cells transfected with IGF2BP3 eukaryotic expression plasmid increased and the apoptosis rate decreased. [ABSTRACT FROM AUTHOR]
- Published
- 2023
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