八月札水提物对胶质瘤细胞EMT的作用机制研究

doi:10.3969/j.issn.1006-6233.2022.02.05

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Abstract Objective: To investigate the effect and mechanism of Akebia trifoliata Koidz aqueous extract on epithelial-mesenchymal transformation of glioma cell line SHG-44 through long non-coding RNA(LncRNA) tumor susceptibility gene(CASC11)/microRNA-641 signaling axis. Methods: SHG-44 cells were cultured in medium containing 0, 30, 60, 90, 120 and 180 μg/mL Akebia trifoliata Koidz aqueous extract, respectively, and the cell survival rate was detected by MTT assay after 24 h. SHG-44 cells were randomly divided into control group, Akebia trifoliata Koidz aqueous extract group (treated with 90μg/mL Akebia trifoliata Koidz aqueous extract for 24h), NC group (transfected with NC for 24h), CASC11 mimic group (transfected with CASC11 mimic for 24h) and Akebia trifoliata Koidz aqueous extract +CASC11 mimic group (after transfecting with CASC11 mimic for 24h, treated with 90μg/mL Akebia trifoliata Koidz aqueous extract for 24h). Dual luciferin reporter gene was used to detect the targeting relationship between CASC11 and miR-641, the expression levels of CASC11 and miR-641 were detected by qRT-PCR, the cell migration ability was detected by scratching experiment, the cell invasion ability was detected by Transwell experiment, and epithelial cadherin (E-cad), nerve cadherin (N- cad) and Vimentin levels were detected by Western blot assay. Results: With the increase of aqueous extract concentration, the cell survival rate decreased in a dose dependent manner (P<0.05). Compared with the control group, the CASC11 expression level, the percentage of wound healing area, the number of invasive cells and the relative expression levels of N-cad and Vimentin proteins were decreased, while the miR-641 expression level and the relative expression level of E-cad protein were increased in Akebia trifoliata Koidz aqueous extract group(P<0.05). Compared with the Akebia trifoliata Koidz aqueous extract group, the expression level of CASC11, the percentage of wound healing area, the number of invasive cells and the relative expression levels of N-cad and Vimentin were increased in the aqueous extract group +CASC11 mimic group, while the expression level of miR-641 and the relative expression level of E-cad protein were decreased (P<0.05). Compared with NC group, the expression level of CASC11, the percentage of wound healing area, the number of invasive cells and the relative expressions of N-cad and Vimentin in CASC11 mimic group were increased, while the expression level of miR-641 and the relative expression level of E-cad protein were decreased (P<0.05). Compared with CASC11 mimic group, the expression level of CASC11, the percentage of scratch healing area, the number of invasive cells and the relative expression levels of N-cad and Vimentin in the group of Akebia trifoliata Koidz aqueous extract +CASC11 mimic were decreased, while the expression level of miR-641 and the relative expression level of E-cad protein were increased (P<0.05). Conclusion: The inhibitory effect of Akebia trifoliata Koidz aqueous extract on the epithelial-mesenchymal transformation of glioma cell line SHG-44 may be achieveded by decreasing the expression of CASC11 while increasing the expression of miR-641.

Key words： Glioma Epithelial-mesenchymal transformation Akebia trifoliata Koidz aqueous extract CASC11/miR-641

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http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2022.02.05 OR http://www.hbyxzzs.cn/EN/Y2022/V28/I2/201

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