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目的:观察消癌解毒方含药血清对人肝癌细胞SMMC-7721细胞中蛋白表达变化的影响。方法:选择人肝癌SMMC-7721细胞作为实验对象,消癌解毒方含药血清作用人肝癌SMMC-7721细胞24h后,采用TMT肽段标记,质谱鉴定分析,运用IPA软件对结果进行生物信息学分析,观察用药前后各组蛋白质表达的差异,应用Western blot法检测Cytochrome C、Caspase-3、Cleaved-Caspase-3蛋白的表达。结果:共鉴定出3 025个蛋白点,消癌解毒方各剂量组与模型组相比差异表达明显的蛋白423个,其中消癌解毒方高剂量组负调控蛋白55个,正调控蛋白85个;消癌解毒方中剂量组负调控蛋白33个,正调控蛋白50个;消癌解毒方低剂量组负调控蛋白36个,正调控蛋白49个;顺铂组负调控蛋白38个,正调控蛋白77个。这些差异表达蛋白涉及脂质代谢、凋亡、氧化应激等方面。信号通路分析涉及Granzyme A Signaling、TR/RXR Activation等通路。Western blot结果显示消癌解毒方处理人肝癌SMMC-7721细胞24h后可引起Cytochrome C蛋白被释放,Cleaved-Caspase-3蛋白表达量上调,Caspase-3蛋白表达量下调。结论:消癌解毒方作用于人肝癌SMMC-7721细胞有多靶点,多中心的调控作用,其中激活线粒体凋亡相关蛋白可能是其抑制肿瘤生长,诱导肿瘤凋亡的途径之一。
Objective: To observe the effect of Xiaoyan Jiedu Fang containing serum on the protein expression of human hepatoma SMMC-7721 cells. Methods: Human hepatocellular carcinoma SMMC-7721 cells were selected as experimental subjects. Human hepatocellular carcinoma SMMC-7721 cells were treated with Xiaojian Jiedu decoction for 24 h. The results were analyzed by mass spectrometry (TMT) and identified by bioinformatics analysis using IPA software The difference of protein expression in each group before and after treatment was observed. Western blot was used to detect the expression of Cytochrome C, Caspase-3 and Cleaved-Caspase-3. Results: A total of 3 025 protein spots were identified. There were 423 differentially expressed proteins in each dose group of Xiaoyao Jiedu Fang compared with the model group. Among them, there were 55 negative regulators and 55 positive regulators The negative control protein of Xiaowangjiedu Fang middle dose group had 33 negative regulators and 50 positive regulators, while the negative control Xiaowangjiedu Fang low dose of 36 negative regulators, positive regulators of 49 and negative regulators of cisplatin 38 positive regulators 77 proteins. These differentially expressed proteins involved in lipid metabolism, apoptosis, oxidative stress and so on. Pathway analysis involves access to Granzyme A Signaling, TR / RXR Activation and more. Western blot results showed that Cytochrome C protein was released by Xiaojian Jiedu Fang treatment of human hepatocellular carcinoma SMMC-7721 cells for 24h, the expression of Cleaved-Caspase-3 protein was up-regulated and the expression of Caspase-3 protein was down-regulated. Conclusion: Xiaojian Jiedu Fang can inhibit the proliferation of human hepatocellular carcinoma SMMC-7721 cells and induce the apoptosis of SMMC-7721 cells. The activation of mitochondrial apoptosis-related proteins may be one of the pathways of inhibiting tumor growth and inducing tumor apoptosis.