复方蜥蜴散基于HIF-1α介导PI3K/AKT通路干预裸鼠胃癌浸润转移

Based on HIF-1α mediating PI3K/AKT pathway of the compound lizard powder intervention in invasion and metastasis of nude mice gastric cancer

目的 研究复方蜥蜴散基于缺氧诱导因子(HIF)-1α介导磷脂酰肌醇3激酶/蛋白激酶B(PI3K/AKT)信号通路干预裸鼠胃癌移植瘤模型上皮间质转化(EMT)的作用,探讨其干预胃癌浸润转移机制。方法 选取120只雄性SPF级4周龄BALB/c裸鼠,采用人胃癌SGC-7901细胞株右腋下皮下注射制备裸鼠胃原位癌种植转移模型,随机分为模型对照组、华蟾素对照组、5-FU对照组、复方蜥蜴散低、中、高剂量组。各组用灌胃或尾静脉注射等方法做相应药物干预治疗6 w后取胃癌组织,TUNEL检测胃癌细胞凋亡,免疫组化法及Western印迹检测胃癌细胞HIF-1α、PI3K、p-AKT、AKT及EMT相关蛋白表达。结果 与模型对照组比较,各药物干预组细胞凋亡率、E-cadherin表达水平显著升高,HIF-1α、PI3K、p-AKT、Integrin-β3相关蛋白表达水平显著降低(P<0.05);与华蟾素、5-Fu阳性对照组比较,复方蜥蜴散不同剂量组细胞凋亡率、E-cadherin表达水平显著升高,HIF-1α、PI3K、p-AKT、Integrin-β3相关蛋白表达水平显著降低(P<0.05);与复方蜥蜴散低、中剂量组比较,高剂量组细胞凋亡率显著升高,HIF-1α、PI3K、p-AKT、Integrin-β3相关蛋白表达水平差异有统计学意义(P<0.05);各组AKT表达水平差异无统计学意义(P>0.05)。结论 复方蜥蜴散能够通过解毒活血通络,改善细胞缺血缺氧微环境,诱导HIF-1α介导PI3K/AKT信号通路,进而影响EMT相关蛋白表达而抑制胃癌细胞浸润转移。

Objective To study the effect of hypoxia inducible factor(HIF)-1α mediating the phosphatidylinositol-3 kinase/protein kinase B(PI3K/AKT) signal pathway on the epithelial-mesenchymal transformation(EMT) of nude mice gastric cancer transplanted tumor model with the compound lizard powder, and explore its intervention mechanism.Methods 120 male SPF grade with 4-week-old BALB/c nude mice were selected, and the model of situ gastric carcinoma in nude mice was prepared by subcutaneous injection of human gastric cancer SGC-7901 cell line into the right axilla.Rats were randomly divided into model control group, cinobufagin control group, 5-FU control group, low, medium and high dose groups of the compound lizard powder.After 6 weeks of intervention treatment with corresponding drugs by gavage or tail vein injection, gastric cancer tissue was taken, TUNEL was used to detect gastric cancer cell apoptosis, immunohistochemistry and Western blot were used to detect gastric cancer cells HIF-1α, PI3K, p-AKT, AKT and EMT-related protein expression.Results Compared with the model control group, the apoptosis rate and the expression level of E-cadherin in each drug intervention group were significantly increased, and HIF-1α, PI3K,p-AKT,Integrin-β3 related proteins expression level were significantly decreased(P<0.05);compared with the cinobufagin and 5-Fu positive control groups, the apoptosis rate and the expression level of E-cadherin in different dose groups of the compound lizard powder were significantly increased, and HIF-1α,PI3K,p-AKT,Integrin-β3 related proteins expression levels were significantly decreased(P<0.05);compared with the low and medium dose groups of the compound lizard powder, the apoptosis rate of the high dose group was significantly higher, and HIF-1α,PI3K,p-AKT,Integrin-β3 related proteins expression level had significant difference(P<0.05);there was no significant difference in AKT expression level among the groups(P>0.05).Conclusions The compound lizard powder could improve the microenvironment of cell ischema and hypoxia by detoxifying and activating blood, induce HIF-1α mediating PI3K/AKT signal pathway, and then affect the expression of EMT-related protein to inhibit the invasion and metastasis of gastric cancer cells.