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目的探讨中药复方解毒散结方对肺癌转移的作用及其机制,为临床运用解毒散解方治疗肺癌,尤其是为防止肺癌术后的复发转移提供依据。方法体外实验采用MTT、划痕、粘附、转移实验来明确解毒散结方抗肺腺癌细胞株A549增殖和侵袭转移的能力。体内实验通过构建裸小鼠人肺癌术后转移模型,将裸鼠随机分为4组:模型组、解毒散结方组、紫杉醇组、解毒散结方联合紫杉醇组,每组10只,观察解毒散解方对裸鼠体质量、肺部肿瘤转移数量、体积和无病生存期(DFS)的影响,并应用免疫组化观察半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)、血管内皮生长因子(VEGF)蛋白的表达。结果 MTT、体外划痕、粘附、转移实验显示在一定浓度范围内解毒散结方可抑制A549细胞增殖、迁移和转移的作用,并能降低其细胞间的粘附能力,且呈现出剂量依赖效应,与正常对照组相比较,差异具有统计学意义(P<0.05)。体内研究发现,解毒散结方组体质量呈不断上升趋势,从术后第3周开始与模型组和紫杉醇组比较,差异具有统计学意义(P<0.05,P<0.01),而且解毒散结方组裸鼠肺部肿瘤转移数量明显低于模型组,差异具有统计学意义(P<0.05),解毒散结方组裸鼠无病生存期也明显高于模型组,差异具有统计学意义(P<0.05)。此外,解毒散结方能促进Caspase3表达的上调和VEGF表达的下调,与模型组相比差异具有统计学意义(P<0.05),且解毒散结方联合紫杉醇组优于解毒散结方组,差异具有统计学意义(P<0.05)。结论解毒散结方可抑制A549肺腺癌细胞的迁移和转移并降低其细胞间的粘附能力,同时,解毒散结方可以延长裸鼠无病生存期,并上调Caspase-3的表达,降低VEGF的表达,提示解毒散结方可以控制肺癌转移,其抗转移的机制可能与促进肿瘤细胞凋亡、抗肿瘤血管生成相关。
Objective To investigate the effect of traditional Chinese medicine Jiedu Sanjie Decoction on metastasis of lung cancer and its mechanism for the clinical application of Jiedu Sanjie for the treatment of lung cancer, especially to prevent lung cancer recurrence and metastasis. Methods MTT, scratch, adhesion and metastasis experiments were performed in vitro to clarify the ability of anti-proliferation and invasion and metastasis of lung adenocarcinoma cell line A549. Nude mice were randomly divided into 4 groups: model group, Jiedu Sanjiefang group, paclitaxel group and Jiedu Sanjiefang combined with paclitaxel group, with 10 mice in each group. Detoxification The effect of Sanjie Fang on nude mice weight, the number of lung tumor metastasis, volume and disease-free survival (DFS), and the expression of caspase-3, Expression of vascular endothelial growth factor (VEGF) protein. Results MTT, in vitro scratch, adhesion and metastasis experiments showed that Jiedu Sanjie can inhibit the proliferation, migration and metastasis of A549 cells in a certain concentration range, and reduce the adhesion between A549 cells in a dose-dependent manner Effect, compared with the normal control group, the difference was statistically significant (P <0.05). In vivo study showed that the quality of Jiedu Sanjie Decoction group showed an increasing trend, and the difference was statistically significant (P <0.05, P <0.01) compared with model group and paclitaxel group from the third week after operation, The number of lung metastases in the untreated group was significantly lower than that in the untreated group (P <0.05), and the disease-free survival of the nude mice in the Jiedu Sanjie group was also significantly higher than that in the untreated group (P <0.05) P <0.05). In addition, Jiedu Sanjie can promote the up-regulation of Caspase3 expression and down-regulation of VEGF expression compared with the model group (P <0.05), and Jiedu Sanjie combined with paclitaxel group is superior to Jiedu Sanjiefang group, The difference was statistically significant (P <0.05). Conclusion Jiedu Sanjie Decoction can inhibit the migration and metastasis of A549 lung adenocarcinoma cells and reduce the cell adhesion ability. At the same time, Jiedu Sanjie can prolong the disease-free survival and up-regulate the expression of Caspase-3 in nude mice, VEGF expression, suggesting that Jiedu Sanjie can control lung cancer metastasis, the anti-metastasis mechanism may be related to promoting tumor cell apoptosis, anti-tumor angiogenesis.