中肺合剂抗小鼠Lewis肺癌肺转移作用及相关机制的实验研究

Anti-metastasis Effect and Mechanism of Zhongfei Mixture in the Mice Bearing Lewis Lung Carcinoma

目的观察中肺合剂对小鼠Lewis肺癌抗肿瘤肺转移的作用,并探索其相关机制。方法采用Lewis肺癌移植瘤C57BL/6小鼠模型观察中肺合剂抗肿瘤肺转移作用,并采用荧光定量QRT-PCR法测定中肺合剂对Lewis肺癌小鼠肿瘤组织基质金属蛋白酶(MMP)-9表达的影响,采用EnVision法检测肿瘤组织血管内皮细胞生长因子(VEGF)和CD34蛋白的表达情况,并计数微血管密度(MVD)。结果中肺合剂低、中、高剂量组(12.7,25.4,50.8g.kg-1)和环磷酰胺(CTX)阳性对照组对移植性Lewis肺癌小鼠的肺转移抑制率为16.7%、45.8%、19.4%和80.6%,以中剂量组肺转移抑制作用最佳。中肺合剂能下调Lewis肺癌小鼠肿瘤组织MMP-9、VEGF蛋白的表达水平,并降低肿瘤组织MVD。MMP-9、VEGF表达和MVD之间呈正相关。结论中肺合剂对小鼠Lewis肺癌具有显著的抗肿瘤肺转移作用,以中剂量组最佳。其作用机制可能与该药能降低肿瘤组织MMP-9、VEGF蛋白的表达水平,并降低肿瘤组织MVD有关。

OBJECTIVE To investigate the anti-metastasis effect and mechanism of Zhongfei Mixture in vivo.METHODS The mouse model of C57BL/6 bearing Lewis lung carcinoma was established to investigate the anti-pulmonary metastasis of Zhongfei Mixture.Further,the expressions of MMP-9 was detected by real-time chain reaction（RT-PCR）and the expression of VEGF,CD34 protein and the microvessel density（MVD）were detected in tumor tissues by EnVision two-step immuno-histochemistry staining.RESULTS The low,middle and high dose of Zhongfei Mixture groups（12.7,25.4,50.8 g·kg-1）as well as CTX（30 mg·kg-1）positive control group significantly inhibited pulmonary metastasis in the mice bearing Lewis lung carcinoma.The inhibitory rates were 16.7%,45.8%,19.4% and 80.6%,respectively,and the middle dose group displayed the most potent anti-pulmonary metastasis effect in the three groups of Zhongfei Mixture.Zhongfei Mixture down-regulated the expression of MMP-9,VEGF and CD34 protein in carcinoma tissues from the mice bearing Lewis lung carcinoma,compared with saline control group,the positive expression rate of the protein in middle and high dose groups was significantly different（P0.01）.There was a significantly positive correlation among MMP-9,VEGF and MVD.CONCLUSION Zhongfei Mixture was an effective anti-tumor agent with anti-metastasis effect in vivo,and the effect of the moderate dose group was optimum.The molecular mechanism of anti-tumor activity could be concerned with down-regulation the MMP-9,VEGF and CD34 in tumor tissues.