基于悬液微珠抗体芯片技术的肝癌和大肠癌术后“同病异证”“异病同证”分析

Analysis on ‘same disease but different syndromes’ and ‘same syndrome exist in different disease’ of post-operation of liver cancer and colorectal cancer based on multiplex biometric immunoassay technology

目的:揭示肝癌和大肠癌术后"同病异证""异病同证"的特征性差异表达的细胞因子。方法:运用悬液微珠抗体芯片技术,定量检测分析证候间45种细胞因子的差异表达谱;运用ELISA对TGF-β1进行定量检测,并对显著差异因子进行验证。结果:通过统计分析发现,IP-10、RANTES、MIP-1β、IL-18、IL-1RA以及IFN-γ在肝癌术后,RANTES、BDNF、PDGF-BB、IL-9以及FGF-2在肠癌术后的肝肾阴虚证、脾虚证、湿热证和隐证中表达差异有统计学意义(P<0.05)。与隐证相比,肝癌和大肠癌术后肝肾阴虚证、脾虚证和湿热证各具有不同的差异表达细胞因子谱。肝癌和大肠癌术后肝肾阴虚证共同的特异性细胞因子为RANTES,脾虚证共同的特异性细胞因子为TGF-β1,但湿热证中未发现共有的异病同证因子。对肝癌和大肠癌术后的共有显著差异细胞因子进行拟合和ROC分析,发现肝肾阴虚证的AUC值为0.837,脾虚证的AUC值为0.871,湿热证的AUC值为0.833,隐证的AUC值为0.936。生物学通路分析发现,Cytokine-cytokine receptor interaction、Cytosolic DNA-sensing pathway、Influenza A和Toll-like receptor signaling pathway等信号通路与肝癌术后同病异证相关;MAPK signaling pathway、Cytokine-cytokine receptor interaction以及Melanoma信号通路与肠癌术后同病异证相关。结论:肝癌和大肠癌术后肝肾阴虚证、脾虚证和湿热证有特异性细胞因子,这些细胞因子的变化可能与其证候的形成有关。

Objective: To explore the cytokines with characteristic differential expression in ‘same disease but different syndromes' and ‘same syndrome exist in different disease' of post-operation of liver cancer and colorectal cancer. Methods: The differential expression profiles of 45 cytokines were analyzed and tested quantificationally by using multiplex biometric immunoassay technology. The expression of TGF-β1 was tested by using ELISA, and the significant difference factors were verified. Results: It found that the expression of IP-10, RANTES, MIP-1β IL-18, IL-1RA, and IFN-γ was statistically significant in liver cancer after operation with syndrome of yin deficiency of liver and kidney, syndrome of spleen deficiency, syndrome of dampness-heat, and hidden syndrome(P<0.05), and the expression of RANTES, BDNF, PDGF-BB, IL-9, and FGF-2 was statistically significant in intestinal cancer after operation with syndrome of yin deficiency of liver and kidney, syndrome of spleen deficiency, syndrome of dampness-heat, and hidden syndrome(P<0.05). Compare with hidden syndrome, post-operation of liver cancer and intestinal cancer with syndrome of yin deficiency of liver and kidney, syndrome of spleen deficiency, syndrome of dampness-heat have their own differentially expressed cytokine profile. The common specific cytokine of post-operation of liver cancer and colorectal cancer with syndrome of yin deficiency of liver and kidney is RANTES, and the common specific cytokine of syndrome of spleen deficiency is TGF-β1, but there is not a common cytokine of same syndrome exist in different disease in syndrome of dampness-heat. Through the analysis of matching and ROC, it found that the AUC values of syndrome of yin deficiency of liver and kidney, syndrome of spleen deficiency, syndrome of dampness-heat and hidden syndrome were 0.837, 0.871, 0.833, and 0.936 respectively. The analysis of biologic pathway showed that the signaling pathways as Cytokine-cytokine receptor interaction, Cytosolic DNA-sensing pathway, Influenza A, and Toll-like receptor signaling pathway were closely related with same disease but different syndromesof post-operation of liver cancer, and the signaling pathways as MAPK signaling pathway, Cytokine-cytokine receptor interaction, and Melanoma were closely related with same disease but different syndromesof postoperation of intestinal cancer. Conclusion: The syndrome of yin deficiency of liver and kidney, syndrome of spleen deficiency, and syndrome of dampness-heat of post-operation of intestinal cancer and liver cancer have their own specific cytokines, and the changes in these cytokines may be related to the formation of their syndrome.