基于WAVE3调控和代谢组学的黄芪-乳香配伍对卵巢癌耐药及侵袭能力作用的机制研究

In clinic, traditional chinese medicine for invigorating blood circulation and eliminating stasis (DRIBCAES) Occupy an important position in drugs of cure neoplasm, but previous studies had query whether it induced resistance and neoplasm metastasis. Our previous study have shown that when drug for supplement qi compatibility used with hemorheologic agent, it not only eliminate possible negative effect of DRIBCAES especially drug for disintegrating blood stasis generated, but also synergistic interaction and delay metastasis. We adopt the supplement qi and invigorate blood representative drugs in treatment of tumor, Olibanum and Astragali Radix, to observe the impact on sensibility of ovarian cancer resistance and invasion and metastasis, from the angle of cell experiment and model of transplanted tumor of ovarian cancer. we introduced LC/TOFMs and PCR to processing theory of Chinese medicine field innovatively for the first time. we discuss the influences on inhibition ratio and transport rate of transplanted tumor of ovarian cancer after WAVE3 silence, using siRNA technique to silence WAVE3 gene. Meanwhile, we detect the expression of correlation factor of resistance P-gp, MRP, BCRP, LRP and imcorrelation factor of transplant MMP-2、VEGF、FGF2、COX-2, utilizing immunohistochemisty and Real time-PCR . We used biological information database to analyze the change rule of metabolic component by means of the overall identification and taget analysis and aimed to preliminary explore the mechanism of inversing tumor drug resistance and inhibiting invasion and metastasis of Olibanum compatibility with Astragali Radix. This providing a scientific basis for enrichment and development of the compatibility theory of Traditional Chinese Medicine, guiding clinical safe and effective usage.

中医临床肿瘤用药中，活血化瘀药占有重要地位，但已有的研究质疑其是否会导致耐药及肿瘤转移。项目前期研究表明：补气活血药配伍使用，不仅能消除活血化瘀药尤破血药带来的负面效应，且能协同增效及延缓转移。本项目对具中医治疗肿瘤特色的补气活血药黄芪及乳香配伍，从细胞、卵巢癌移植瘤模型及代谢组分等观察其对卵巢癌耐药敏感性及侵袭转移的影响。项目创新性地将LC/TOF/MS、PCR等前沿技术首次引入中医药理论研究领域。以siRNA技术沉默WAVE3基因，探讨配伍对卵巢癌移植瘤的抑制率及转移率的影响，以PCR检测耐药相关因子P-gp, MRP, BCRP, LRP及转移相关因子MMP-2、VEGF、FGF2、COX-2的表达，通过整体辨识和靶标分析，利用生物信息数据库解析代谢组分的变化规律，初步探寻黄芪、乳香配伍逆转肿瘤耐药及抑制侵袭转移机理，为丰富和发展中药配伍理论，指导临床安全、有效用药提供科学依据。

中医临床肿瘤用药中，活血化瘀药占重要地位，但已有研究质疑其是否会导致肿瘤细胞耐药及增加肿瘤转移。本项目对具有中医治疗肿瘤特色的补气活血药黄芪及乳香配伍，从细胞、移植瘤模型及代谢组学等观察其对卵巢癌耐药逆转及侵袭转移的影响。项目创新性地将基因敲除、基因过表达、LC/TOF/MS、PCR、WB等前沿生物学技术首次引入中医药理论研究领域。以siRNA技术沉默WAVE3基因、细胞转染技术过表达WAVE3 pcDNA3.1质粒，探讨补气活血配伍对卵巢癌移植瘤的抑制率及转移率的影响，以WB检测转移相关因子MMP-9、MMP-2、VEGF、 NF-KB、COX-2、pP38、P38的表达，通过整体辨识和靶标分析，利用生物信息数据库解析代谢组分的变化规律，探寻黄芪与乳香配伍抑制卵巢癌侵袭转移的分子机理，为丰富和发展中药配伍理论提供理论基础，并为临床卵巢癌的抗转移的治疗提供新的治疗方案和科学依据。项目研究表明，补气活血配伍不仅能消除活血化瘀药尤其破血药带来的负面效应，且能协同增效及延缓肿瘤转移。