槲皮素对宫颈癌裸鼠模型肿瘤生长及其对乙酰肝素酶表达的关系研究

Study on relationship of quercetin on cervical carcinoma in nude mice model of tumor growth and the expression of heparanase

目的研究槲皮素对宫颈癌生长的作用,及其对乙酰肝素酶表达的关系,初步探讨槲皮素抑制宫颈癌作用的可能机制。方法建立宫颈癌裸鼠模型,将植瘤成功的30只裸鼠进行如下随机分组:25 mg/kg槲皮素组、50 mg/kg槲皮素组、100 mg/kg槲皮素组和PI-88阳性对照组及空白对照组;观察各组透射电镜下血管内皮细胞超微结构的变化;以Western blot蛋白免疫印记技术检测癌瘤中乙酰肝素酶蛋白质水平改变,将空白组做为灰度参照,计算出各组间相对灰度值,进行统计学分析。结果 50 mg/kg、100mg/kg槲皮素组及PI-88组较空白组中宫颈癌生长体积小(P<0.05);50 mg/kg组可见肿瘤组织中血管内皮细胞和线粒体肿胀等改变,100 mg/kg组可见膜性结构的消失及胞质边集等改变,PI-88组可见管腔闭锁,液泡状滑面内质网和死亡的细胞;50 mg/kg组和100mg/kg组宫颈癌植瘤瘤体组织中乙酰肝素酶表达较空白对照组减少(P<0.05),PI-88组较50 mg/kg组和100 mg/kg组乙酰肝素酶表达减少(P<0.01),而25 mg/kg组较空白对照组差异无统计学意义。结论 50、100 mg/kg槲皮素可抑制宫颈癌细胞生长,其可导致宫颈癌植瘤瘤体内血管内皮细胞细胞超微结构的改变。槲皮素对宫颈癌的抑制作用可能与降低乙酰肝素酶蛋白表达有关。

Objective To explore effect of quercetin on growth of cervical cancer,and its relationship to expression heparanase,and to investigate the possible mechanism of the inhibition effect of quercetin on cervical cancer. Methods He La cells were injected into nude mouse to establish the model of cervical cancer. 30 nude mice with palpable tumors were randomly assigned to five groups： 25,50,100 mg / kg quercetin groups,PI-88 positive group and blank control group. Transmission electron microscope was used to observe the ultrastructural changes of the vascular endothelial cells in each group.The expression of heparanase protein was detected by Western blot. Used blank control group for grey reference,relative grey value of each group was calculated for the further statistical analysis. Results 50,100 mg / kg quercetin groups and group of PI-88 made the volume of tumor tissues smaller. In50 mg / kg quercetin group,swelling of VEC and mitochondria in tumor tissues was observed. In 100 mg / kg quercetin group,the disappearance of membraneaceous structures was found. In PI-88 positive group,the atresia of lumen,vacuole-shaped smooth endoplasmic reticulum and a dead cell were observed. 50 and 100 mg / kg quercetin groups decreased the heparanase protein levels significantly（ P〈0. 05）. Furthermore,PI-88 positive group could decrease the heparanase protein levels significantly（ P〈0. 05）,compared to 50 and 100 mg / kg quercetin groups. However,the difference between the25 mg / kg quercetin group and blank control group showed no statistical significance. Conclusion The growth of cervical cancer could be inhibited by 50,100 mg / kg quercetin. Meanwhile,the ultrastructural changes of tumor endothelial cells could be induced by 50,100 mg / kg quercetin. These changes may relate with the decrease of heparanase protein level.