乳腺癌是女性最常见的恶性肿瘤，主要的转移途径是淋巴转移, 淋巴管生成在淋巴转移过程中起着至关重要的作用，决定着乳腺癌分期和治疗方案的选择。长期以来人们对乳腺癌的研究一直把注意力放在血管生成上，对淋巴管生成与淋巴转移的问题没有深入研究。

随着促淋巴管生长因子和一些特异性淋巴管内皮标记物的发现，淋巴管生成和淋巴转移逐渐成为新的研究热点[1]。在众多淋巴管生成因子中，最具特点的淋巴管生长因子是血管内皮生长因子C/D(vascular endothelial growth factor-C/D，VEGF-C/D)及血管内皮生长因子受体-3(vascularendothelial growth factor receptor-3，VEGFR-3)，VEGF-C和VEGF-D通过与其受体VEGFR-3的结合促进淋巴管的生成。本研究采用人乳腺癌高转移细胞株MDA-MB-435S乳房原位移植BALB/c Nu/Nu裸鼠，建立自发性转移模型，以VEGF-C/D为切入点，研究乳腺癌组织中VEGF-C/D mRNA及VEGFR-3 mRNA的表达，探讨中药对乳腺癌高转移裸鼠模型淋巴管生成相关因子的影响，寻找中药干预淋巴转移的作用机制。

材料与方法

1. 动物及细胞株　BALB/c Nu/Nu裸鼠，雌性，6周龄，体质量(20±2)g，购自广西医科大学实验动物中心，许可证号：

SCXK(桂)2009-0002，于SPF条件下饲养，适应环境1周后进行实验。人乳腺癌MDA-MB-435S细胞株购于南京凯基生物科技发展有限公司。

2. 试剂　供试RPMI-1640培养液、胎牛血清购自Gibco公司(美国)、一步法RT-PCR试剂盒购自北京天根生化科技有限公司，Trizol试剂购自Invitrogen公司，RT-PCR引物由Invitrogen(上海)公司合成，化学发光剂购自Santa Cruz公司，预染色的蛋白Marker购自Solarbio公司。

3. 仪器　CM1900型冰冻切片机(德国LEICA公司)，3K30型高速冷冻离心机(德国Sigma公司)，MCO -15AC型二氧化碳培养箱(日本三洋)，FACSCalibur型流式细胞仪(美国B-D公司)，Lambda Bio 20型紫外分光光度计(美国Gene公司)，iCycler型梯度PCR仪(美国Bio-Rad公司)，Ge1Doc2000型凝胶电泳成像分析系统(美国Bio-Rad公司)，稳压稳流电泳仪及水平电泳槽(北京六一仪器厂)，电热恒温干热培养箱(天津中环实验电炉仪器厂)，电子天平(上海方瑞仪器有限公司)。

4. 药物　乳康饮制剂由黄芪30g，茯苓15g，青皮12g，柴胡9g，莪术9g，薏苡仁15g组成，药物购于泰山医学院附属医院中药房，经泰山医学院药学院鉴定，由药学院药剂学实验室制成Effects of Rukangyin on -s of VEGF-C/D, VEGFR-3 mRNA in breast cancertissue of nude miceSUN Xi-bo, LI Xiang-qi

( Affi liated Hospital of Taishan Medical University, Taian 271000, China )

Abstract: -ive: To observe the influence of traditional Chinese medicine compound Rukangyin (RKY) on -s of VEGF-C/D, VEGFR-3 mRNA in transplantation tumor of mice breast cancer, and to discuss the inhibition

mechanism of breast cancer -stasis. 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 -stasis models. They

were randomly divided into six groups, 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 mice in each group. Normal saline was given to mice in the model control group

at the daily dose of 0.4mL by gastrogavage. 5-FU was given to mice in the 5-FU control group at the daily dose of 30mg/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 90g/kg

by gastrogavage, respectively. 5-FU 30mg·kg-1·d-1 (by peritoneal injection)+RKY 45g·kg-1·d-1(by gastrogavage) were given to mice

in the medium dose RKY+5-FU group. All medications were carried out once daily for 6 successive weeks. The tumor volume,

the tumor inhibition ratio and the inhibition ratio of axillary lymph node -stasis were detected after medications treatment. The

- of VEGF-C/D and 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 (P

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