藏红花素抑制AKT和STAT3的活化对黑色素瘤A375细胞裸鼠移植瘤生长、运动及多器官病变的影响  

Effect of crocin inhibiting the activation of AKT and STAT3 on the growth, movement and multiple organ lesions of transplanted melanoma A375 cells in nude mice

作　　者：李媛[1] 李娜[1] 付文静[1] 舒春梅[1] 张玉杰[1] 马冲[2] Li Yuan;Li Na;Fu Wenjing(Dept of Dermatology and Venereology,The Affiliated Hospital of Binzhou Medical University,Binzhou 256600)

机构地区：[1]滨州医学院附属医院皮肤性病科,滨州256600 [2]滨州医学院附属医院结直肠肛门疝与腹壁外科,滨州256600

出　　处：《安徽医科大学学报》2020年第7期1073-1079,共7页Acta Universitatis Medicinalis Anhui

基　　金：山东省医药卫生科技发展计划项目(编号:2017WS545)。

摘　　要：目的探索藏红花素对黑色素瘤A375细胞生长、运动和对肿瘤引起的多器官病变的影响是否存在AKT和STAT3信号通路的参与,以及该通路相关蛋白的表达情况。方法建立黑色素瘤A375细胞裸鼠皮下移植瘤模型,将小鼠随机分成4组,分别用0、20、50、100 mg/kg浓度的藏红花素和生理盐水对瘤体进行局部注射,每次0.1 ml,观察瘤体变化。免疫组织化学法检测Ki67、Caspase-3的含量以及血管内皮细胞生长因子(VEGF)和波形蛋白(Vimetin)的表达;RT-PCR检测生长标志物Survivin、Bcl-2/Bax基因的表达;HE染色观察肝、肺、肾的组织病理形态;免疫印迹法检测运动和上皮-间质转化(EMT)标志物E-cadherin、N-cadherin以及Fibronectin的蛋白表达以及蛋白激酶B(AKT)和信号转导和转录激活因子3(STAT3)的磷酸化情况。结果与对照组比较,藏红花素低、中、高剂量组黑色素瘤体积减小,重量降低(P<0.05);Ki67、VEGF、Vimentin阳性细胞数减少(P<0.05),Caspase-3阳性细胞增加(P<0.05);Survivin mRNA的表达降低(P<0.05);Fibronectin蛋白表达水平降低(P<0.05)。与对照组比较,藏红花素中、高剂量组黑色素瘤中Bcl-2/Bax mRNA的表达降低(P<0.05);E-cadherin、N-cadherin蛋白表达水平降低(P<0.05);AKT和STAT3的磷酸化水平降低(P<0.05)。结论藏红花素通过抑制AKT和STAT3的磷酸化,减轻肝、肺和肾组织的病理形态学改变,抑制A375黑色素瘤的生长和运动,表明藏红花素对A375黑色素瘤有缓解作用,可减轻肝、肺、肾的组织损伤。藏红花素可以作为有效的调节剂用于预防和治疗黑色素瘤。Objective To investigate the effect of crocin on the growth, exercise and multi-organ injury of melanoma A375 cells, as well as the expression of proteins related to the AKT and STAT3 signaling pathways.Methods A subcutaneously transplanted tumor model of A375 nude mice was established. The mice were randomly divided into 4 groups, and the tumor was injected with 0, 20, 50, 100 mg/kg of crocin and normal saline respectively, and 0.1 ml each time to observe the changes of the tumor. Immunohistochemistry was used to detect the levels of Ki67,Caspase-3 and the expression of VEGF and Vimetin. The expression of Survivin and Bcl-2/Bax genes was detected by RT-PCR. The histopathological morphology of liver,lung and kidney was observed by HE staining. Western blot was used to detect the expression of motility and EMT markers E-cadherin,N-cadherin and Fibronectin,as well as phosphorylation of AKT and STAT3. Results Compared with the control group,the volume and weight of melanomas in the low,medium and high dose groups of Crocin decreased( P < 0. 05);the number of Ki67,VEGF and Vimentin positive cells decreased( P < 0. 05),while the number of Caspase-3 positive cells increased( P < 0. 05);the expression of Survivin mRNA decreased( P < 0. 05);the expression of Fibronectinprotein decreased( P < 0. 05).Compared with the control group,the expression of Bcl-2/Bax mRNA in the middle and high dose groups decreased( P < 0. 05);the expression levels of E-cadherin and N-cadherin decreased( P < 0. 05);the phosphorylation levels of AKT and STAT3 decreased( P < 0. 05);Crocin alleviated the tissue damage of liver,lung and kidney. Conclusion By inhibiting the phosphorylation of AKT and STAT3,Crocin can reduce the pathomorphological changes of liver,lung and kidney tissues,and inhibit the growth and movement of A375 melanoma,suggesting that saffron has a relieving effect on A375 melanoma. Saffron can be used as an effective and safe non-toxic regulator to prevent and treat melanoma.

关 键 词：藏红花素 AKT STAT3 黑色素瘤 器官病变 

分 类 号：R739.5[医药卫生—肿瘤]