Zhang Q, Zhang W T, Wu S S, Yuan J J, Tian L, Liu Y Y, Zuo W L, Song Y P, Zhou K S
Department of Hematology, the Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital, Zhengzhou 450008, China.
Zhonghua Xue Ye Xue Za Zhi. 2018 Jul 14;39(7):579-583. doi: 10.3760/cma.j.issn.0253-2727.2018.07.010.
To explore the expression and prognostic significance of miR-223 in patients with mantle cell lymphoma (MCL) and to investigate the possible mechanism. 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. 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 1 244.1±1 935.2, <0.001). The lower expression of miR-223 was inversely correlated with high-risk mantle international prognostic index (=0.001), elevated LDH (=0.001), ECOG score ≥2 (=0.035). ②Using the median relative expression level of miR-223 as the cutoff value, 21 MCL patients were divided into high-expression group (=10) and low-expression group (=11) and found that the high-expression group had a significantly superior OS (median OS: 36 12 months, =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, <0.001), manifested by lower proportion of cells in G2/M phase (<0.001) and increased apoptosis (<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 (=-0.81, <0.001). 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.
探讨微小RNA-223（miR-223）在套细胞淋巴瘤（MCL）患者中的表达及预后意义，并研究其可能机制。本研究纳入21例新诊断的伴有骨髓侵犯的MCL患者，20例健康供者作为正常对照。采用实时定量聚合酶链反应（RQ-PCR）检测miR-223和SOX11 mRNA的表达水平。采用细胞计数试剂盒-8（CCK-8）法和流式细胞术分析构建的miR-223过表达MCL细胞系Granta519细胞的增殖、细胞周期及凋亡情况。采用蛋白质免疫印迹法检测SOX11蛋白表达水平。通过双荧光素酶报告基因实验验证miR-223的靶基因。①21例新诊断的MCL患者中，男性15例，女性6例，中位年龄58（37~72）岁。与健康供者相比，MCL患者miR-223表达水平显著下调（14.7±10.5比244.1±1935.2，<0.001）。miR-223低表达与高危套细胞国际预后指数（=0.001）、乳酸脱氢酶升高（=0.001）、东部肿瘤协作组（ECOG）评分≥2（=0.035）呈负相关。②以miR-223相对表达水平的中位数为截断值，将21例MCL患者分为高表达组（=10）和低表达组（=11），发现高表达组总生存期显著更长（中位总生存期：36比12个月，=0.021）。③体外实验结果显示，与对照组相比，miR-223过表达的Granta519细胞增殖受到抑制（96小时时抑制最显著，<0.001），表现为G2/M期细胞比例降低（<0.001），凋亡增加（<0.001），且Granta519细胞中SOX11蛋白表达水平显著低于对照组。④miR-223可抑制SOX11的3'非翻译区，且MCL患者中miR-223表达水平与SOX11 mRNA水平显著负相关（=-0.81，<0.001）。MCL中miR-223表达受抑制且与不良临床结局相关，这可能归因于其直接靶向SOX11。
