miR-335调控Survivin和NF-κB表达对B细胞淋巴瘤细胞侵袭、迁移和凋亡的影响

Effects of miR-335 on Survivin and NF-κB expressions on invasion,migration and apoptosis of B-cell lymphoma cells

目的:探讨miR-335调控Survivin和NF-κB表达对B细胞淋巴瘤细胞侵袭、迁移和凋亡的影响。方法:取对数生长期的B细胞淋巴瘤细胞株SU-DHL-4,分为对照组(不做任何处理)、空转染组(转染空载体)和过表达组(转染miR-335)。转染48 h后采用RT-PCR和Western blot分别检测各组细胞miR-335水平和Survivin、NF-κB蛋白的相对表达量。采用划痕实验和Transwell侵袭实验检测细胞迁移和侵袭能力。流式细胞仪检测细胞凋亡率。结果:与对照组相比,空转染组miR-335表达水平、Survivin蛋白表达水平、细胞迁移率、侵袭细胞数和细胞凋亡率差异无统计学意义(P>0.05),过表达组miR-335表达水平和细胞凋亡率明显升高(P<0.05),Survivin和NF-κB蛋白表达水平、细胞迁移率和侵袭细胞数明显降低(P<0.05)。与空转染组相比,过表达组miR-335表达水平和细胞凋亡率明显升高(P<0.05),Survivin和NF-κB蛋白表达水平、细胞迁移率和侵袭细胞数明显降低(P<0.05)。结论:miR-335可通过调控Survivin表达,下调NF-κB信号通路,诱导肿瘤细胞凋亡,并抑制B细胞淋巴瘤细胞的迁移和侵袭。

Objective:To investigate the effects of miR-335 on invasion,migration and apoptosis of B cell lymphoma cells by regulating expressions of Survivin and NF-κB protein.Methods:B cell lymphoma cell strain SU-DHL-4 in logarithmic growth stage were divided into control group(without any treatment),empty transfection group(transfected with empty vector)and overexpression group(transfected with miR-335).RT-PCR and Western blot were used to detect relative expressions of miR-335 and Survivin,NF-κB protein in each group 48 h after transfection.Scratch test and Transwell invasion test were used to detect cell migration and invasion ability.Flow cytometry was used to measure apoptosis rate.Results:Compared with control group,expression of miR-335,expression levels of Survivin protein,cell mobility,cell numbers of invasion and cell apoptosis rate in empty transfection group have no obvious difference(P>0.05),while expression of miR-335 and cell apoptosis rate in overexpression group were increased signi-ficantly(P<0.05),expression levels of Survivin and NF-κB protein,cell mobility and cell numbers of invasion were decreased significantly(P<0.05).Compared with empty transfection group,expression level of miR-335 and cell apoptosis rate in overexpression group were increased significantly(P<0.05),expression levels of Survivin and NF-κB protein,cell mobility and cell numbers of invasion were decreased significantly(P<0.05).Conclusion:miR-335 can induce apoptosis of tumor cells and inhibit migration and invasion of B-cell lymphoma cells by regulating expressions of Survivin protein,reduce NF-κB signaling pathway.