乳康饮对裸鼠乳腺癌自发性转移模型淋巴管生成的干预作用

Intervention of Rukangyin on the Lymphangiogenesis in Breast Cancer Spontaneous Metastasis Nude Mouse Model

目的观察复方中药乳康饮对乳腺癌移植瘤小鼠淋巴管生成和淋巴结转移的影响,探讨其抗肿瘤转移机制。方法 30只裸鼠乳腺原位种植人乳腺癌细胞株MDA-MB-435S,建立乳腺癌移植瘤自发性转移模型,随机分成6组,分别为模型组,5-氟尿嘧啶(5-FU)对照组,乳康饮小、中、大剂量组以及乳康饮加5-FU组,每组5只。模型组给予生理盐水0.4 mL/d灌胃;5-FU对照组给予30 mg/(kg.d)5-FU腹腔注射;乳康饮小、中、大剂量组分别给予乳康饮18、45、90 g/(kg.d)灌胃;乳康饮加5-FU组给予5-FU 30 mg/(kg.d)腹腔注射,同时给予乳康饮45 g/(kg.d)灌胃,均每天1次,连续6周。用药后检测肿瘤体积、抑瘤率和腋窝淋巴结转移抑制率;免疫组化法检测乳腺癌组织微淋巴管密度(lymphatic microvessel density,LMVD)和血管内皮生长因子C(vascular endothelial growth factor-C,VEGF-C)、血管内皮生长因子受体-3(vascular endothelial growth factor re-ceptor-3,VEGFR-3)的表达。结果与模型组比较,各用药组肿瘤体积明显减小,VEGF-C和VEGFR-3表达水平显著下调,LMVD明显降低,差异均有统计学意义(P<0.05,P<0.01);其中以乳康饮加5-FU组抑瘤率及腋窝淋巴结转移抑制率最高,LMVD最低,与其他用药组比较,差异均有统计学意义(P<0.05,P<0.01)。结论乳康饮可能通过干预VEGF-C及VEGFR-3的表达,影响淋巴管的生成,抑制乳腺癌淋巴转移。

Objective To observe the effects of Rukangyin（RKY） on the lymphangiogenesis and lymph node metastasis of breast cancer transplantation tumor mice,thus exploring its anti-tumor metastasis mechanisms.Methods Human breast cancer cell line MDA-MB-435S were in situ implanted into the mammary fat pad of 30 female nude mice to establish breast cancer transplantation tumor spontaneous metastasis model.They were randomly divided into six groups,i.e.,the model control group,the 5-FU control group,the small,medium,large dose RKY groups,and the medium dose RKY＋5-FU group,5 in each.Normal saline was given to mice in the model control group at the daily dose of 0.4 mL/kg by gastrogavage.5-FU was given to mice in the 5-FU control group at the daily dose of 30 mg/kg by peritoneal injection.RKY was given to mice in the small,medium,large dose RKY groups at the daily dose of 18,45,and 90 g/kg by gastrogavage.5-FU 30 mg/kg（by peritoneal injection）＋RKY 45 g/（kg·d）（by gastrogavage） was given to mice in the medium dose RKY＋5-FU group.All medication was carried out once daily for 6 successive weeks.The tumor volume,the tumor inhibition ratio,and the inhibition ratio of axillary lymph node metastasis were detected after medication.The lymphatic microvessel density（LMVD） and expressions of vascular endothelial growth factor-C（VEGF-C） and vascular endothelial growth factor receptor-3（VEGFR-3） of the breast cancer tissues were detected using immunohistochemical assay.Results Compared with the model control group,the tumor volume was markedly reduced in the small,medium,large dose RKY groups,and the medium dose RKY＋5-FU group,the expressions of VEGF-C and VEGFR-3 were significantly down-regulated and LMVD were obviously lowered,showing statistical difference（P0.05,P0.01）.The inhibition rates of tumor and axillary lymph node metastasis were highest and the LMVD was the lowest in the medium dose RKY＋5-FU group,showing statistical difference when compared with other medication groups（P0.05,P0.01）.Conclusion RKY might inhibit the lymph node metastasis of breast cancer possibly through intervening the expressions of VEGF-C and VEGFR-3,and suppressing lymphangiogenesis.