基于消退素调控炎症相关通路探讨片仔癀治疗肝癌的作用机制

Inflammation is closely related to the occurrence and development of cancers. Recently, it has been found that residual cancer cells can stimulate tumor growth by inducing inflammatory response after conventional treatment. Resolvins are endogenous lipid derived mediators, which can suppress the growth and metastasis of cancer by inhibiting inflammatory response and promoting the solution of inflammation. Pien Tze Huang (PZH) is a traditional Chinese medicine, which has been confirmed having good anti-inflammatory and anti-cancer effect in clinic. Our previous study found that Pien Tze Huang could suppress cancer progression, inhibit the inflammatory signaling pathway and promote the resolvins generation in hepatocellular carcinoma-bearing mice, suggesting that its anti-cancer effect might be through regulating the production of resolvins and their related pathways. Therefore, in this study, we plan to establish two kinds of hepatocellular carcinoma mice models: orthotopic transplantation tumor models of hepatocellular carcinoma and metastases models, detect the changes of inflammation-related signal pathways in mice models after PZH treatment by RNA sequence technology, and analyze the metabolism characteristics of resolvins by metabolic analysis. Then, biological information technology will be used to analyze the correlation between RNA sequence data and metabolomic data. The inflammatory signal pathways and target proteins involved in the PZH treating hepatocellular carcinoma will be predicted. Finally, experiments will be performed to verify these signal pathways and target proteins. This research will clarify the action mechanism of PZH regulating resolvins-involved inflammatory signal pathway in hepatocellular carcinoma, which will provide scientific evidence for PZH being used in the prevention and treatment of hepatocellular carcinoma.

炎症和肿瘤的发生发展密切相关，近来研究发现常规治疗后残余的肿瘤细胞能通过诱导机体炎症反应来刺激肿瘤继续生长。消退素是一种内源性脂质衍生的介导子，能通过抑制炎症反应、促进炎症解决，进而抑制肿瘤生长和转移。片仔癀是我国传统中成药，临床证实其具有良好的抗炎及抗肿瘤作用。我们前期研究发现：片仔癀可以抑制荷肝癌小鼠肿瘤生长、抑制炎症信号通路活化并促进消退素的产生，提示其可能通过调控消退素及相关通路来发挥抗瘤作用。据此，本研究拟建立原位肝移植瘤及肝转移瘤小鼠模型，采用RNA测序检测片仔癀对荷瘤鼠炎症信号通路的影响，并借助代谢组学平台技术分析药物干预后荷瘤鼠血液消退素的代谢特征；进而通过生物信息学技术对代谢组学结果与RNA测序结果进行关联分析，推测药物作用通路；最后借助免疫学及分子生物学技术对所预测的通路及靶蛋白进行实验验证，明确片仔癀调控消退素及其相关炎症通路的分子机制，为片仔癀防治肝癌提供科学依据。

慢性炎症与肝癌的发生、发展及转移密切相关。消退素能通过抑制炎症反应、促进炎症解决，进而抑制肿瘤生长和转移。片仔癀是我国传统中成药，临床证实其具有良好的抗炎及抗肿瘤作用，但其作用特点及机制不明。我们前期研究提示片仔癀可能通过调控消退素及相关通路来发挥抗瘤作用。据此，本项目以消退素及炎症通路为切入点，采用多组学联合生物信息学方法对片仔癀治疗肝癌的作用特点及分子机制开展了一系列研究工作，结果发现：片仔癀对肝移植瘤小鼠具有明确的治疗效果，可以抑制肿瘤生长，减少荷瘤小鼠血清中促炎细胞因子的表达，改善肿瘤炎症微环境。片仔癀可以调控SPMs及其前体脂肪酸代谢，特别是17(S)-HDHA、19(20)-EpDPE、D系列消退素。被调控的SPMs及其前体脂肪酸通过参与KDR、CD5、GSK3A等分子及NF-κB和PI3K/AKT信号通路网络的调控发挥抗炎作用。此外，片仔癀治疗肝癌主要通过参与炎症相关通路的调控，而索拉非尼则主要通过参与癌症相关通路的调控。片仔癀和索拉非尼均可延缓肿瘤细胞周期的进展，其中片仔癀在细胞周期中诱导G2/M的阻滞，索拉非尼则主要引起G1期转化延迟。进一步的免疫学及分子生物学实验也证实了上述结果。综上，本项目证明了片仔癀对肝移植瘤小鼠具有确切的治疗效果；明确了片仔癀治疗肝癌的作用机制与调控消退素及其相关炎症通路相关，这为片仔癀治疗肝癌提供了更丰富的实验依据。此外，本项目还发现片仔癀与索拉非尼在治疗肝癌过程中作用机制存在不同，即片仔癀在抗炎方面具有优势，以上结论可为索拉非尼联合片仔癀治疗肝癌提供实验证据。