PKCδ表达与弥漫大B细胞淋巴瘤疗效和预后的相关性及其机制研究

Diffuse large B cell lymphoma is the most common type of lymphoma. After 6-8 cycles of R-CHOP regimen chemotherapy, most patients can be cured with a 5-year survival rate of 70%. In spite of this, some patients had poor response to initial treatment, or recurrence in the short term, with poor prognosis. Therefore, it is very important to search for markers related to curative effect and prognosis, and to explore new targeted therapies. PKCδ is a serine/threonine kinase involved in cell proliferation, growth and apoptosis. Previous studies have found that in diffuse large B-cell lymphoma, the prognosis of PKCδ is relatively poor. This study proposed by increasing sample size to verify the relationship between PKCδ and curative effect and prognosis of, in the experiments in vivo and in vitro, using RNA interference by PKCδ express, according to the drug sensitive test confirmed that PKCδ associated with lymphoma curative effect indeed. By using RNA-seq technology and western blot method,clarifying detailed mechanism of PKCδ's involvement in drug resistance , and find a targeted drug that could reverse the drug resistance of PKCδ. Finally, it can improve the treat efficacy of diffuse large B cell lymphoma and improve it's prognosis.

弥漫大B细胞淋巴瘤是最常见的一种淋巴瘤亚型。经过6-8个周期的R-CHOP方案化疗，大部分患者可以治愈，5年生存率可达到70%。尽管如此，仍有部分患者对初始治疗反应差，或者短期内复发，预后极差。因此，寻找与疗效和预后相关的标志物，对于预测疗效，探索新的靶向治疗非常重要。PKCδ是一种丝氨酸/苏氨酸蛋白激酶，参与细胞增殖、生长和凋亡等功能。前期研究发现在弥漫大B细胞淋巴瘤中，PKCδ高表达患者预后相对较差。本研究拟通过扩大样本量验证PKCδ与疗效和预后的关系，在体内外实验中，采用RNA干扰下调PKCδ表达，根据药敏试验证实PKCδ与淋巴瘤疗效确实有关。通过RNA-seq技术、western blot方法阐明PKCδ参与耐药的详细机制，并找到一种可以逆转PKCδ耐药的靶向药物。最终达到提高弥漫大B细胞淋巴瘤疗效，改善预后。

弥漫大B细胞淋巴瘤是最常见的一种淋巴瘤亚型，部分患者对初始治疗反应差，或者短期内复发，预后极差。本研究通过扩大样本量通过免疫组化验证PKCδ与疗效和预后的关系，发现PKCδ在DLBCL中的表达水平与患者预后呈显著负相关；在体内外实验中，采用RNA干扰下调PKCδ表达，根据药敏试验分析PKCδshRNA、抑制剂联合利妥昔单抗对淋巴瘤疗效的影响，发现PKCδ基因用shRNA 沉默、利妥昔单抗及化疗药物干预以后，细胞存活率显著降低，细胞增殖受到抑制，凋亡细胞显著增多，细胞周期S期显著缩短，G0/G1期延长。RNA-seq测序发现，PKCδ基因用shRNA 沉默以后，共有2988个基因显著差异表达，其中1400个上调，1588个下调，KEGG和GO富集分析显示，差异表达的2988个基因主要富集在AKT信号通路，MAPK信号通路，NFKB信号通路等通路中。裸鼠成瘤实验发现，PKCδ基因敲除、利妥昔单抗和PKCδ抑制剂治疗后均能显著抑制瘤块生长，且PKCδ抑制剂和利妥昔单抗联用时治疗效果更好。本次研究表明，PKCδ高表达是DLBCL的不良预后因素，低表达时可显著增强利妥昔单抗免疫治疗效果，PKCδ是弥漫大B细胞淋巴瘤药物治疗的潜在分子靶点，对其进行干预有望提高弥漫大B细胞淋巴瘤疗效，改善患者预后。