miR-223靶向调控SOX11在套细胞淋巴瘤中的预后意义

Prognostic significance of miRNA-223 targeting SOX11 in mantle cell lymphoma

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2l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miR223、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Westernblot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（1244．1土1935．2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（96h时最明显，P〈0．001）、处于G2／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SOXll而发挥作用。

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 ① Of 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 intemational 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 significantlysuperior OS （median OS： 36 vs 12 months, P = 0.021）. @In 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 SOXll 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 tar-etin- SOX 11.