摘要
目的:探讨绞股蓝皂苷调节音猬因子(Shh)/胶质细胞瘤转录因子1(Gli1)信号通路对乳腺癌大鼠肿瘤生长和转移的影响。方法:选取40只SD雌性大鼠,通过皮下注射大鼠乳腺癌SHZ-88细胞构建乳腺癌大鼠模型,并随机分为SHZ-88组、绞股蓝皂苷-75组(75 mg/kg绞股蓝皂苷)、绞股蓝皂苷-150组(150 mg/kg绞股蓝皂苷)和绞股蓝皂苷-150+Shh/Gli1信号通路激活剂嘌吗啡胺(PM)组(150 mg/kg绞股蓝皂苷+10 mg/kg PM),每组10只。绞股蓝皂苷以灌胃的方式给药,体积为0.5 ml;灌胃后间隔30 min腹腔注射PM;SHZ-88组给予相同剂量的0.9%氯化钠溶液;每日1次,连续21 d。治疗结束收集大鼠血液并记录肿瘤的重量和体积;ELISA检测乳腺癌大鼠血清中癌胚抗原(CEA)、糖类抗原(CA15-3)、雌激素受体(ER)和孕激素受体(PR)浓度;HE染色观察肿瘤和肝组织病理学变化;免疫组织化学染色检测肿瘤组织细胞增殖核抗原(Ki-67)、ER和PR阳性率;TUNEL染色检测肿瘤细胞凋亡率;RT-qPCR检测Shh/Gli1信号通路基因转录水平;Western blot检测Shh/Gli1信号通路及转移相关蛋白表达。结果:与SHZ-88组比较,绞股蓝皂苷-75组和绞股蓝皂苷-150组大鼠肿瘤肿瘤和体积减小,肿瘤细胞排列较为有序且细胞形态完整,肝组织细胞坏死得到改善,转移灶面积占比减少,血清CEA、CA15-3、ER和PR水平降低,肿瘤组织中Ki-67、ER、PR阳性率及Shh、PTCH1、SMO、Gli1 mRNA和Shh、PTCH1、SMO、Gli1、波形蛋白(Vimentin)蛋白表达水平降低,肿瘤细胞凋亡率和肿瘤组织上皮钙黏蛋白(E-cadherin)蛋白表达水平升高(均P<0.05)。与绞股蓝皂苷-150组比较,绞股蓝皂苷-150+PM组大鼠肿瘤重量和体积增加,肿瘤细胞变形严重,排列不齐,肝组织病变加剧,转移灶面积占比升高,血清CEA、CA15-3、ER和PR水平升高,肿瘤组织中Ki-67、ER、PR阳性率及Shh、PTCH1、SMO、Gli1 mRNA和Shh、PTCH1、SMO、Gli1、Vimentin蛋白表达水平升高,肿瘤细胞凋亡率和肿瘤组织E-cadherin蛋白表达水平降低(均P<0.05)。结论:绞股蓝皂苷可能通过阻断Shh/Gli1信号通路抑制乳腺癌大鼠肿瘤生长和转移。
Objective:To investigate the effects of gypenosides on tumor growth and metastasis in breast cancer rats by regulating the Sonic hedgehog(Shh)/glioma-associated oncogene homolog 1(Gli1)signaling pathway.Methods:Forty female SD rats were selected and subcutaneously injected with SHZ-88 rat breast cancer cells to establish a breast cancer rat model.The rats were randomly divided into four groups:SHZ-88 group,gypenosides-75 group(75 mg/kg gypenosides),gypenosides-150 group(150 mg/kg gypenosides),and gypenosides-150+Shh/Gli1 signaling pathway activator purmorphamine(PM)group(150 mg/kg gypenosides+10 mg/kg PM),with 10 rats in each group.Gypenosides were administered orally at a volume of 0.5 ml,followed by intraperitoneal injection of PM 30 minutes later.The SHZ-88 group received the same volume of 0.9%saline.Treatments were given once daily for 21 consecutive days.After treatment,blood samples were collected,and tumor weight and volume were recorded.ELISA was used to detect serum levels of carcinoembryonic antigen(CEA),carbohydrate antigen 15-3(CA15-3),estrogen receptor(ER)and progesterone receptor(PR).HE staining was used to observe pathological changes in tumor and liver tissues.Immunohistochemistry was used to detect the positive rates of Ki-67,ER and PR in tumor tissues.TUNEL staining was used to assess tumor cell apoptosis.RT-qPCR was used to measure the mRNA expression levels of Shh/Gli1 signaling pathway-related genes.Western blot was used to detect the expression of proteins related to the Shh/Gli1 signaling pathway and metastasis.Results:Compared with the SHZ-88 group,the gypenosides-75 and gypenosides-150 groups showed reduced tumor weight and volume,more orderly tumor cell arrangement with intact morphology,improved hepatic necrosis,decreased metastatic lesion area,lower serum levels of CEA,CA15-3,ER and PR,reduced positive rates of Ki-67,ER and PR in tumor tissues,decreased mRNA and protein expression levels of Shh,PTCH1,SMO and Gli1,as well as Vimentin protein in tumor tissues,and increased tumor cell apoptosis rate and E-cadherin protein expression in tumor tissues(all P<0.05).Compared with the gypenosides-150 group,the gypenosides-150+PM group showed increased tumor weight and volume,severe tumor cell deformation and disarray,aggravated liver tissue lesions,increased metastatic lesion area,elevated serum levels of CEA,CA15-3,ER and PR,increased positive rates of Ki-67,ER and PR in tumor tissues,elevated mRNA and protein expression levels of Shh,PTCH1,SMO,Gli1,as well as Vimentin protein in tumor tissues,and decreased tumor cell apoptosis rate and E-cadherin protein expression in tumor tissues(all P<0.05).Conclusion:Gypenosides may inhibit tumor growth and metastasis in breast cancer rats by blocking the Shh/Gli1 signaling pathway.
作者
王瑞芹
赵龙挺
连婧
马丽娜
武鹏鹏
WANG Ruiqin;ZHAO Longting;LIAN Jing;MA Lina;WU Pengpeng(Department of Traditional Chinese Medicine,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China;Department of Traditional Chinese Medicine,Yangquan Institute for Cancer Prevention and Treatment Research,Yangquan 045000,China;Department of Pathology,Shanxi Province Cancer Hospital,Shanxi Hospital Affiliated to Cancer Hospital,Chinese Academy of Medical Sciences,Cancer Hospital Affiliated to Shanxi Medical University,Taiyuan 030013,China)
出处
《陕西医学杂志》
2026年第2期166-174,共9页
Shaanxi Medical Journal
基金
山西省基础研究计划项目(202103021223445)。
