miR-34a靶向CD47对弥漫性大B细胞淋巴瘤SU-DHL-2细胞生物学行为的影响

Effect of miR-34a targeting CD47 on biological behavior of diffuse large B-cell lymphoma cells SU-DHL-2

目的:探究miR-34a靶向CD47对弥漫性大B细胞淋巴瘤(DLBCL)细胞生物学行为的影响。方法:体外培养人DLBCL细胞SU-DHL-2,随机分为对照组、miR-34a mimics阴性对照组、miR-34a mimics组,CCK-8和流式细胞术分别测定各组SU-DHL-2细胞活力、凋亡率;细胞划痕和Transwell侵袭实验检测SU-DHL-2细胞迁移、侵袭能力;qRT-PCR检测SU-DHL-2细胞miR-34a和CD47 mRNA表达;Western blot检测凋亡相关蛋白Bcl-2、Bax、上皮-间质转化(EMT)相关蛋白Vimentin、E-cadherin和CD47蛋白表达;双荧光素酶报告基因实验检测miR-34a对CD47的靶向调节关系。结果:与对照组相比,miR-34a mimics组SU-DHL-2细胞凋亡率、miR-34a表达、E-cadherin及Bax蛋白表达明显升高(P<0.05),细胞活力、迁移细胞数、侵袭细胞数、CD47 mRNA表达及CD47、Vimentin、Bcl-2蛋白表达显著降低(P<0.05),miR-34a mimics阴性对照组SU-DHL-2细胞各指标差异无统计学意义(P>0.05)。miR-34a可靶向下调CD47表达。结论:miR-34a可靶向下调CD47表达,抑制DLBCL细胞增殖,诱导其凋亡,降低其侵袭转移能力。

Objective:To investigate effect of miR-34a targeting CD47 on biological behavior of diffuse large B-cell lymphoma(DLBCL)cells.Methods:Human DLBCL cells SU-DHL-2 were cultured in vitro and randomly divided into control group,miR-34a mimics negative control group and miR-34a mimics group.Activity and apoptosis rate of SU-DHL-2 cells were measured by CCK-8 and flow cytometry in each group;migration and invasion of SU-DHL-2 cells were detected by cell scratch test and Transwell invasion test;qRT-PCR was used to detect expressions of miR-34a and CD47 mRNA in SU-DHL-2 cells;expressions of apoptosis related proteins Bcl-2,Bax,epithelial-mesenchymal transition(EMT)related proteins Vimentin,E-cadherin and CD47 were detected by Western blot;double luciferase reporter gene assay was used to detect targeting regulatory relationship between miR-34a and CD47.Results:Compared with control group,apoptosis rate,miR-34a expression,E-cadherin and Bax protein expressions were significantly increased in miR-34a mimics group(P0.05).miR-34a could targeting down-regulate expression of CD47.Conclusion:miR-34a can targeting down-regulate expression of CD47,inhibit proliferation,induce apoptosis and decrease invasion and metastasis of DLBCL cells.