Objective: To explore the expression and prognostic significance of miR-223 in patients with mantle cell lymphoma (MCL) and to investigate the possible mechanism. Methods: Twenty-one newly diagnosed MCL patients with bone marrow involvement were enrolled in the present study, 20 healthy donors as normal control. The expression level of miR-223 and SOX11 mRNA was determined by RQ-PCR. CCK-8 and flow cytometer assays were used to analyze cell proliferation, cell cycle and apoptosis of the constructed miR-223 overexpressing MCL cell line, Granta519 cells. SOX11 protein expression level was determined by Western blot. The target gene of miR-223 was confirmed by dual luciferase reporter assay. Results: ①Of the 21 newly diagnosed MCL patients, 15 were male and 6 female, the median age was 58 (37-72) years. The expression level of miR-223 was significantly down regulated in MCL patients compared with that of healthy donors (14.7±10.5 vs 1 244.1±1 935.2, P<0.001). The lower expression of miR-223 was inversely correlated with high-risk mantle international prognostic index (P=0.001), elevated LDH (P=0.001), ECOG score ≥2 (P=0.035). ②Using the median relative expression level of miR-223 as the cutoff value, 21 MCL patients were divided into high-expression group (n=10) and low-expression group (n=11) and found that the high-expression group had a significantly superior OS (median OS: 36 vs 12 months, P=0.021). ③In vitro results showed that compared with the control group, the proliferation of miR-223 overexpressed Granta519 cells was inhibited (the most significant reduction on 96h, P<0.001), manifested by lower proportion of cells in G2/M phase (P<0.001) and increased apoptosis (P<0.001), and the expression level of SOX11 protein in Granta519 cells was significantly lower than that of the control group. ④miR-223 could inhibited the 3' untranslated region of SOX11, and the expression level of miR-223 was significantly negatively correlated with mRNA level of SOX11 in MCL patients (r=-0.81, P<0.001). Conclusions: The expression of miR-223 was repressed in MCL and was associated with poor clinical outcomes, which may be probably attributed to its direct targeting SOX11.
目的： 探讨miR-223在套细胞淋巴瘤（MCL）患者中的表达及预后意义，并探讨可能的作用机制。 方法： 以21例骨髓受累初治MCL患者为研究对象，以20例健康正常供者为正常对照，采用RQ-PCR法检测miR-223、SOX11 mRNA表达水平。构建过表达miR-223的MCL细胞系Granta519细胞，采用CCK8法和流式细胞术检测其增殖、周期和凋亡水平，采用Western blot法检测其SOX11蛋白表达水平，并利用双荧光素酶报告基因实验验证miR-223的靶基因。 结果： ①21例MCL患者中，男15例，女6例，中位年龄58（37~72）岁，其中17例为中高危组。与正常对照组比较，MCL组患者miR-223表达水平显著下调（1 244.1±1 935.2对14.7±10.5，P<0.001），且其低表达与MCL的IPI评分高危组（P=0.001）、LDH升高（P=0.001）、ECOG评分≥2分（P=0.035）等高危临床特征相关。②以患者组miR-223中位表达水平为阈值，将患者分为高表达组（10例）和低表达组（11例），生存分析结果显示前者的总生存时间较后者延长（36个月对12个月，P=0.021）。③体外实验结果显示，与对照组比较，过表达miR-223的Granta519细胞增殖受抑（96 h时最明显，P<0.001）、处于G(2)/M期的细胞明显减少（P<0.001）、细胞凋亡比例增加（P<0.001）；Granta519细胞的SOX11蛋白表达水平较对照组明显降低。④miR-223可抑制SOX11的3′非翻译区；MCL患者的miR-223与SOX11 mRNA表达水平呈明显负相关（r=-0.81，P<0.001）。 结论： miR-223在MCL患者中低表达，且与不良预后相关，机制上可能通过靶向SOX11而发挥作用。.
Keywords: Mantle cell lymphoma; MicroRNAs; SOX11 transcription factor.