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目的观察补肾健脾方对人肝癌高转移原位移植瘤裸鼠切除术后血管生成拟态(Vasculogenic Mimicry,VM)形成以及肺转移的影响。方法建立HCCLM3原位荷瘤切除术后复发转移裸鼠模型,分为补肾健脾方组(A组)、索拉非尼组(B组)、联合用药组(C组)、模型对照组(D组),每组7只。术后第3天起,给予相应药物灌胃,共给药28天,每天1次。末次给药后48 h处死裸鼠并取材,采用免疫组化法检测复发瘤体VE-cadherin、Eph A2、PI3K、FAK蛋白表达,计算血管生成拟态密度(Vasculogenic mimicry density,VMD)以及肺转移细胞数。结果 (1)VMD:C组与其余3组比较,VECadherin蛋白阳性面积显著降低(P<0.05)。与D组相比较,A和C组Eph A2蛋白表达显著降低(P<0.05);A、B、C组PI3K、FAK蛋白表达均显著下调(P<0.05)。VMD由低到高依次为C、A、B、D组,与D组比较,下调倍数依次为1.4、1.2、1.1倍,但差异无统计学意义(P>0.05)。(2)肺转移细胞数:与D组比较,A、C组肺转移肿瘤细胞数显著减少(P<0.05)。结论补肾健脾方能有效降低VE-Cadherin、Eph A2、PI3K、FAK蛋白的表达,减少肺部转移;与索拉非尼联用时,对VE-Cadherin、Eph A2蛋白表达抑制尤为显著,能最大程度阻止VM的形成,降低肺部转移。
Objective To observe the effect of Bushen Jianpi Fang on the formation of Vasculogenic Mimicry (VM) and lung metastasis in nude mice after orthotopic transplanted human hepatoma with high metastasis. Methods The model of HCCLM3 tumor recurrence and metastasis in nude mice after resection of tumor was established and divided into three groups: Bushen Jianpi Fang group (A group), sorafenib group (B group), combination therapy group (C group) and model control group D group), 7 in each group. After the first 3 days, given the corresponding drug gavage, administered a total of 28 days, 1 day. The nude mice were sacrificed at 48 h after the last administration, and the expressions of VE-cadherin, EphA2, PI3K and FAK were detected by immunohistochemistry. Vasculogenic mimicry density (VMD) and lung metastasis number. Results (1) Compared with the other three groups, the positive area of VECadherin protein in VMD group was significantly lower (P <0.05). Compared with group D, the expression of EphA2 in group A and group C was significantly decreased (P <0.05). The protein expressions of PI3K and FAK in groups A, B and C were significantly decreased (P <0.05). The order of VMD was C, A, B, D from low to high. Compared with D group, the down-regulation times were 1.4,1.2,1.1 times, but the difference was not statistically significant (P> 0.05). (2) The number of lung metastasis cells: Compared with D group, the number of lung metastasis cells in A and C groups decreased significantly (P <0.05). Conclusion Bushenjianpi Decoction can effectively reduce the expression of VE-Cadherin, EphA2, PI3K and FAK protein and reduce the lung metastasis. When combined with sorafenib, the expression of VE-Cadherin and EphA2 protein is significantly inhibited To a certain extent prevent the formation of VM and reduce lung metastasis.