西黄解毒胶囊调控上皮间质转化抑制三阴性乳腺癌侵袭转移的作用机制

Xihuang Jiedu Capsules Inhibit Invasion and Metastasis of Triple-Negative Breast Cancer by Regulating Epithelial-Mesenchymal Transition

目的 三阴性乳腺癌(Triple negative breast cancer, TNBC)具有高侵袭性、高转移特性,临床预后极差,及早地预防和抑制三阴性乳腺癌转移对此类患者具有重要意义。西黄解毒胶囊为广安门医院院内制剂,临床在各阶段乳腺癌治疗中反馈良好。通过体内外实验观察西黄解毒胶囊对TNBC侵袭转移能力的影响,并探索其可能作用机制。方法 体外用西黄解毒胶囊干预4T1细胞,通过细胞侵袭实验和划痕愈合实验观察西黄解毒胶囊对4T1细胞侵袭和迁移能力的影响;体内通过建立4T1荷瘤小鼠模型,观察西黄解毒胶囊对4T1荷瘤小鼠肺重及肺转移发生率的影响。免疫荧光实验和免疫组化实验分别观察西黄解毒干预后细胞和动物组织中E-Cadherin和N-cadherin蛋白的表达情况,以初步探索其可能作用机制。结果 细胞侵袭实验结果显示,与对照组比较,西黄解毒高、中、低剂量组细胞侵袭个数明显降低,差异有统计学意义(P<0.05)。划痕愈合实验结果显示,与对照组和DMSO组比较,西黄解毒高、中、低剂量组划痕愈合率呈下降趋势,且高剂量组与对照组比较,差异有统计学意义(P<0.05)。与模型组比较,其他各组肺重降低,特别是联合组、化疗组和西黄解毒高剂量组降低最为明显。HE染色结果显示,与模型组比较,西黄解毒高、中、低剂量组显示出一定降低肺转移的趋势,且与化疗组比较,联合组有更好的趋势。免疫荧光检测结果显示与对照组比较,西黄解毒高、中、低剂量组能明显上调E-cadherin、下调N-cadherin蛋白的表达,差异有统计学意义(P<0.01)。免疫组化结果显示,与模型组比较,西黄解毒高、中、低剂量组明显增加E-caherin的表达,降低N-cadherin的表达,差异有统计学意义(P<0.01),且联合组优于化疗组,差异有统计学意义(P<0.01)。结论 西黄解毒能降低三阴性乳腺癌的侵袭转移能力,其作用机制可能与其逆转了上皮间质转化过程有关。

Objective Triple-negative breast cancer(TNBC)has high possibilities of invasion and metastasis and poor prognosis,and thus early prevention and inhibition of TNBC metastasis is of great significance for the patients.Xihuang Jiedu Capsules,an in-hospital preparation of Guang′anmen Hospital,have received good feedback in the clinical treatment of breast cancer at all stages.In this study,the effects of Xihuang Jiedu Capsules on the invasion and metastasis of TNBC were observed through cell and animal experiments,and the possible mechanism was explored.Methods Transwell assay and cell scratching assay were employed to examine the invasion and migration of 4T1 cells treated with Xihuang Jiedu Capsules.Furthermore,the effects of Xihuang Jiedu Capsules on the lung weight and lung metastasis incidence in 4T1 tumor-bearing mice were observed.The expression of E-cadherin and N-cadherin in cell and animal tissues after the intervention was observed by immunofluorescence and immunohistochemical assays,respectively,on the basis of which the possible mechanism was preliminarily explored.Results Compared with the control group,high,medium,and low doses of Xihuang Jiedu Capsules reduced the number of cell invasion(P<0.05).In the cell scratching experiment,high,medium,and low doses of Xihuang Jiedu Capsules decreased the healing rate compared with the control group and DMSO group,and the difference between the high dose group and the control group was statistically significant(P<0.05).Compared with the model group,other groups,especially the groups of combined therapy,chemotherapy,and high-dose Xihuang Jiedu Capsules showed reduced lung weight.The results of hematoxylin-eosin staining showed that compared with the model group,high,medium,and low doses of Xihuang Jiedu Capsules reduced the rate of lung metastasis,and the combined therapy outperformed chemotherapy.The results of immunofluorescence assay showed that compared with the control group,high-,medium-,and low-dose Xihuang Jiedu Capsules up-regulated the expression of E-cadherin and down-regulated the expression of N-cadherin(P<0.01).The immunohistochemical assay results showed that compared with the model group,high-,medium-and low-dose Xihuang Jiedu Capsules up-regulated the expression of E-caherin and down-regulated the expression of N-cadherin(P<0.01),and the combined therapy was superior to the chemotherapy(P<0.01).Conclusion Xihuang Jiedu Capsules can inhibit the invasion and metastasis of TNBC by reversing the epithelial-mesenchymal transition.