基于Aurora激酶A研究消癌平注射液协同化疗药物抗Burkitt淋巴瘤的效应和机制

It was demonstrated in our previous clinical and experimental studies, that Xiaoaiping injection in combination with chemotherapy has shown synergistic inhibition on lymphoma,and Xiaoaiping injection lowered the protein expression of Burrkitt lymphoma cell line Raji. Encouraged by these results, we are trying to explain the mechanism and clarify the targets involved. Aurora–A gene is overexpressed in Burkitt lymphoma cells and could be regulated by Xiaoaiping injection in our preliminary studies. Employing gene silencing or amplifying technique in lymphoma cells to manipulate Aurora–A gene, we will test the corperation effect of Xiaoaiping injection with chemo-regimen in this system and interpretation their role to control and adjust Aurora-A gene. And we will also figure out the relationship of Aurora-A gene regulation with synergistic inhibition by cellular and molecular experiments involving proliferating inhibition, cell cycle arresting, apoptosis induction and chemotherapy sensitization. Based on Aurora –A gene, molecular markers by which Xiaoaiping injection exhibits its efficacy will be explored, thus the important theoretical and realistic significance of reasonable utilization of Xiaoaiping injection and the integral treatment of lymphoma will be presented.

前期临床与实验研究显示，消癌平注射液联合化疗药物对淋巴瘤的治疗具有协同效应，Aurora激酶A在人Burkitt淋巴瘤细胞株Riji细胞中呈过表达，消癌平注射液能够降低其表达水平。本研究在此基础上，拟从消癌平注射液对Riji细胞增殖抑制、细胞周期阻滞、诱导凋亡和化疗增敏等方面明确其与化疗药物的协同作用，通过Aurora激酶A基因转染或沉默技术探讨消癌平注射液协同化疗药物抗Burkitt淋巴瘤的作用靶点，揭示消癌平注射液对化疗药物协同作用与其对Aurora激酶A调控作用的相关关系，明确消癌平注射液是否通过介导Aurora激酶A发挥对化疗药物抗淋巴瘤的协同作用。本项目以新型Aurora激酶A为切入点，探明消癌平注射液抗肿瘤的确切药效分子生物标志物，对消癌平注射液的合理应用与研发及淋巴瘤的中西医结合治疗具有重要的理论意义和现实意义。

Burkitt淋巴瘤（Burkitt lymphoma,BL）是一种高度恶性的淋巴瘤，其治疗仍然存在挑战。消癌平注射液是由传统中药通关藤的提取物，属于新一代单味多组份的广谱抗癌中药注射剂。前期临床研究发现消癌平注射液联合化疗方案可以明显提高B细胞淋巴瘤的缓解率和疾病控制率。为了探索消癌平注射液的作用机制，本研究首先采用RNA干扰技术(siRNA)抑制Raji细胞中Aurora-A的表达，然后通过体内外实验，验证消癌平注射液通过调控Aurora激酶A的表达发挥抗Burkitt淋巴瘤的作用。在体外实验中，MTT、流式检测结果表明，消癌平注射液能够明显抑制siRNA前后Raji细胞的增殖，促进其凋亡，导致细胞发生细胞周期G1期阻滞。在Aurora-A表达正常的情况下，消癌平注射液单独应用以及和化疗药物组合应用在一定程度上均能够抑制细胞周期相关蛋白Cyclin D1和P27 mRNA的表达，抑制BCL-2和C-myc的表达，而上调促凋亡蛋白Bax、Caspase-3、Caspase-8、Caspase-9的水平，能够抑制耐药相关蛋白P-gp、MRP1、BMI1和MDR1 的表达。但是在si-Aurora-A后，无论是消癌平注射液单药还是中西医结合治疗，上述蛋白的表达水平明显呈现明显差异。另外，体内和体外实验表明，消癌平注射液还可以通过靶向Aurora-A表达调控PI3K/AKT信号通路的活性，并增加P53和P21的活性来实现抗淋巴瘤作用。同时，消癌平注射液抗肿瘤血管生成的作用也是其抗淋巴瘤的重要途径。本课题以新型Aurora-A为作用靶点，结合基因调控技术，进一步明确了中药注射剂消癌平注射液的作用机制和靶点，为临床使用消癌平注射液提供有力参考。