扶正抗癌方对Lewis肺癌荷瘤小鼠肺癌基因Bcl-2、抑癌基因Bax及血管内皮生长因子、表皮生长因子受体、金属蛋白酶表达的影响

Effects of Fuzheng Kangai Decoction on Expressions of the Bcl-2 Lung Cancer Gene,Tumor-Suppressor Gene Bax and VEGF,EGFR,MMP in Mice with Lewis Lung Cancer

目的:观察扶正抗癌方对小鼠Lewis肺癌抗转移机制的影响。方法:选取C57BL/6小鼠50只,雌雄各半,每只小鼠腋窝下接种肿瘤细胞0. 2 m L造模,24 h后,完全随机分为对照组、顺铂组、扶正抗癌方低、中、高剂量组,每组10只。分别给药21 d后取出完整肿瘤组织称重计算抑瘤率,同时用实时荧光定量PCR检测Lewis肺癌组织Bcl-2和Bax mRNA的表达,免疫组化检测Lewis肺癌组织基质金属蛋白酶-9(MMP-9)蛋白的表达,用蛋白质印迹法(WB法)检测Lewis肺癌组织血管内皮生长因子(VEGF)、表皮生长因子受体(EGFR)的表达。结果:与模型组小鼠比较,扶正抗癌方药可以呈剂量依赖性抑制Lewis肺癌荷瘤小鼠肿瘤生长,显著升高Lewis肺癌组织中Bax mRNA表达,降低Bcl-2 mRNA、MMP-9蛋白表达,与模型组比较,差异有统计学意义(P <0. 05),其中高剂量组在升高Lewis肺癌组织中Bax mRNA表达、降低Bcl-2 mRNA表达方面与顺铂组比较,差异有统计学意义(P <0. 05)。同时模型组Lewis肺癌组织中VEGF、EGFR表达显著升高,扶正抗癌方药可显著降低Lewis肺癌组织中VEGF、EGFR表达,与模型组比较,差异有统计学意义(P <0. 05),与顺铂组比较,无统计学意义(P> 0. 05)。结论:虽然扶正抗癌方总体上较顺铂作用偏弱,但依然能有效抑制肺癌细胞增殖,抑制肺癌细胞Bcl-2基因的表达,激活Bax的高表达,下调肺癌细胞中VEGF、EGFR、MMP的表达。

Objective:To observe Fuzheng Kangai Decoction on the anti-transfer mechanism of Lewis lung cancer in mice.Methods:A total of 50 C57BL/6 mice,half male and half female,were selected,and each mouse was inoculated with 0.2 mL of tumor cells under the armpit for 24 hours,and then were randomly divided into control group,DDP group,Fuzheng Kangai Decoction low,medium and high dose group,with 10 mice in each group.They were respectively administrated for 21 days,and the complete tumor tissue was taken out for weighing inhibitory rate calculation.The expression of Bcl-2 and Bax mRNA in Lewis lung cancer tissues were detected by Real-time fluorescent quantitative polymerase chain reaction (PCR) detection.The expression of MMP-9 protein in Lewis lung cancer tissues was detected by immunohistochemistry.And the expression of vascular endothelial growth factor (VEGF),epidermal growth factor receptor (EGFR) in Lewis lung cancer tissues was detected by method of western blot (WB).Results:Compared with the model group,Fuzheng Kangai Decoction could inhibit the tumor growth of Lewis lung cancer-bearing mice in a dose-dependent manner,and significantly increase the expressions of Bax mRNA in Lewis lung cancer tissues,and decrease the expression of Bcl-2 mRNA and MMP-9 protein.There were significant differences compared with the model group (P<0.01).The expressions of VEGF,EGFR in Lewis lung cancer tissues was significantly increased in the model group,and Fuzheng Kangai Decoction could significantly reduce the expressions of VEGF,EGFR in Lewis lung cancer tissues.There was a significant difference compared with the model group (P<0.01,0.05).Conclusion:Fuzheng Kangai Decoction can inhibit lung cancer cell proliferation,inhibit the expression of Bcl-2 gene in lung cancer cells,activate the high expression of Bax and down-regulate the expression of VEGF,EGFR and MMP in lung cancer cells.