沉默重组蛋白2对NCI-H1688细胞的抑制作用及其Wnt/β连环蛋白信号通路机制

doi:10.13481/j.1671-587x.20200414

Abstract

Abstract: Objective: To investigate the effect of frequently rearranged in advanced T-cell lymphomas 2 (Frat2) on the proliferation of small cell lung cancer NCI-H1688 cells via Wnt/β-catenin signaling pathway,and to elucidate its possible mechanism. Methods: The human small cell lung cancer NCI-H1688 cells with good growth status and the normal bronchial epithelial-like cells HBE were selected;Western blotting method and Real-Time RT-qPCR were used to detect the expression levels of Frat2 mRNA and protein in the NCI-H1688 cells and the HBE cells.The NCI-H1688 cells were randomly divided into blank control group, negative control group,Frat2-siRNA group and Frat2-siRNA+XAV group.The cells in blank control group were not transfected.The cells in negative control group were transfected with negative control lentivirus,the cells in Frat2-siRNA group were transfected with Frat2-siRNA lentivirus and the cells in Frat2-siRNA+XAV group were transfected with the Frat2-siRNA lentivirus and treated with 4 μmol·L^-1of Wnt signaling pathway inhibitor XAV939.The expression levels of Frat2 protein and mRNA in the cells in various groups were determined by Western blotting method and Real-Time RT-qPCR method.The proliferation activities of the cells in various groups were determined by MTT method.The percentages of cells in different cell cycles were measured by flow cytometry.The expression levels of Frat2, proliferating cell nuclear antigen (PCNA), c-myc, cyclin D1, β-catenin and phosphorylated glycogen synthase kinase 3 (pGSK-3β) proteins in the cells were determined by Western blotting method. Results: Compared with the normal bronchial epithelial-like cells HBE, the levels of Frat2 protein and mRNA in the NCI-H1688 cells were elevated (P<0.01).Compared with blank control group and negative control group, the expression levels of protein and Frat2 mRNA in Frat2-siRNA group were increased (P<0.05).There were no significant difference in the expression levels of Frat2 protein and mRNA in the NCI-H1688 cells between blank control group and negative control group (P>0.05).Compared with blank control group and negative control group, the proliferation activity of the cells in Frat2-siRNA group was decreased (P<0.05), the percentage of cells in G₀/G₁ phase was increased (P<0.05),the percentages of cells in S phase and G₂/M phase were decreased (P<0.05), and the expression levels of PCNA, c-myc, Cyclin D1, β-catenin and pGSK-3β proteins were decreased (P<0.05).Compared with Frat2-siRNA group,the proliferation activity of cells in Frat2 mRNA+XAV group was decreased(P<0.05),the percentage of cells in G₀/G₁ phase was increased (P<0.05), the percentages of cells in S phase and G₂/M phase were decreased (P<0.05),and the expression levels of PCNA, c-myc, Cyclin D1, β-catenin and pGSK-3β proteins were decreased (P<0.05).There were no significant differences in the indexes mentioned above between blank control group and negative control group (P>0.05). Conclusion: Silencing Frat2 can inhibit the proliferation of small cell lung cancer cells, and its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.

Key words: frequently rearranged in advanced T-cell lymphomas 2, Wnt/β-catenin signaling pathway, small cell lung cancer, proliferation, proliferating cell nuclear antigen, cyclin D1, glycogen synthase kinase 3

CLC Number:

R734.2

CHEN Yan, PAN Dianzhu, LIU Zhong, MA Yanmei, ZHANG Lan. Inhibitory effect of sileneing Frat2 on NCI-H1688 cells and its Wnt/β catenin signaling pathway mechanism[J].Journal of Jilin University(Medicine Edition), 2020, 46(04): 751-758.

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Inhibitory effect of sileneing Frat2 on NCI-H1688 cells and its Wnt/β catenin signaling pathway mechanism

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