Peroxiredoxin-6调控Fas/FasL凋亡途径对弥漫性大B细胞淋巴瘤发生、发展的作用

Diffuse large B cell lymphoma (DLBCL) is a common kind of invasive B cell tumor with highly heterogeneity in China. The pathogenesis of DLBCL is not clear and there is lack of effective indicators for its prognosis evaluation. In our preliminary study, we confirmed for the first time that Peroxiredoxin-6 (Prdx6), a new kind of antioxidant, was over-expressed in DLBCL tissues and it was closely related with tumour invasive behaviors. On the basis of published studies and our work, we propose a hypothesis that over-expressed Prdx6 in DLBCL could inhibit Fas/FasL-induced cell apoptosis, thus improving the development and progression of lymphoma. To test this hypothesis, we will continuously detect the expression of Prdx6 in DLBCL tissues by immunohistochemical method and analyze their correlation with the prognosis of DLBCL. In addition, using human diffuse large B cell lymphoma cells, we will explore the role of Prdx6 in the proliferation, apoptosis and progression of lymphoma cells by building Prdx6 RNA interference or over-expression lentivirus vectors and infecting target cells with them to down/up-regulate the expression of Prdx6 in tumour cells, further indicating its effect on the regulation of Fas/FasL-mediated apoptosis signalling pathway. Moreover, by setting up a lymphoma-burdened mice model, we will confirm the relationship between the abnormal expression of Prdx6 and the development and progression of DLBCL in vivo. Our findings would provide a new molecular marker for DLBCL diagnosis and its prognosis evaluation.

弥漫性大B细胞淋巴瘤（DLBCL）是常见的具有高度异质性的侵袭性淋巴瘤，其发生机制不明确且缺乏预后评估有效指标。我们首次证实一种新的抗氧化蛋白Peroxiredoxin-6（Prdx6）在DLBCL中高表达并与侵袭行为相关。在总结前人及自己工作基础上，我们提出假说即Prdx6在DLBCL中高表达，异常表达的Prdx6通过抑制Fas/FasL介导的细胞凋亡途径，从而促进淋巴瘤发生、发展过程。为验证该假说，我们通过病例分析Prdx6异常表达与DLBCL预后相关性；以人DLBCL细胞株为研究对象，构建针对Prdx6基因的慢病毒RNA干扰或过表达载体并感染靶细胞，体外探讨Prdx6基因在淋巴瘤细胞增殖、凋亡及侵袭转移中的作用及其对Fas/FasL凋亡通路的调控；构建淋巴瘤荷瘤裸鼠模型，体内证实Prdx6异常表达及其调控机制与DLBCL发生、发展的关系，为DLBCL诊断及预后评估提供新的分子标记物。

弥漫性大B细胞淋巴瘤（DLBCL）是常见的具有高度异质性的侵袭性淋巴瘤，我们首次证实一种新的抗氧化蛋白Peroxiredoxin-6（Prdx6）在DLBCL中高表达，且与非生发中心组（non-GCB）组、B症状和乳酸脱氢酶（LDH）有关。同时，Prdx6高表达与患者预后密切相关，提示可能对DLBCL的形成以及发生、发展机制具有一定作用。接着，我们以DLBCL细胞株为研究对象，构建针对Prdx6基因的慢病毒RNA干扰或过表达载体并感染靶细胞。由于悬浮细胞感染困难，我们通过增加细胞株类型（OCI-LY1、OCI-LY19、SU-DHL-4、SU-DHL-10、Farage）以及离心法进行操作，最终筛选出OCI-LY1和Farage进行后续实验。考虑到前期靶细胞构建耗费较多时间，我们同时进行Prdx6基因上调和下调对其增殖、凋亡、侵袭、迁移、克隆形成等研究。但从目前的结果来看，两株细胞并不具有侵袭、迁移和克隆形成能力，只有CCK8、凋亡以及CellTiter中少量数据有统计学意义。所以，后续也许可以尝试从凋亡、以及KD6方向入手接着往下做一些研究，但结果具有一定的不确定性。由于课题时间限制以及人员能力问题，目前我们只能研究到这里。