麻疹减毒活疫苗Hu191株对4T1乳腺癌细胞上皮-间质转化、增殖及迁移的影响  

Effects of live attenuated measles vaccine Hu191 strain on epithelial mesenchymal transition,proliferation and migration of 4T1 breast cancer cells

作　　者：姚梦玮 李玉迁 徐庆胜 方小菡 魏洪[1,2] 迟茜文 刘蕊 YAO Mengwei;LI Yuqian;XU Qingsheng;FANG Xiaohan;WEI Hong;CHI Qianwen;LIU Rui(National Experimental Teaching Center,School of Basic Medicine,Guizhou Medical University,Guiyang 550025,Guizhou Province,China;不详)

机构地区：[1]贵州医科大学基础医学国家级实验教学中心,贵州贵阳550025 [2]贵州省普通高校学校病原生物学特色重点实验室,贵州贵阳550025 [3]贵州省人民医院病理科,贵州贵阳550002 [4]贵州中医药大学第二附属医院红岩院区中心实验室,贵州贵阳550003

出　　处：《中国生物制品学杂志》2023年第12期1434-1441,共8页Chinese Journal of Biologicals

基　　金：贵州省卫生健康委科学技术基金(gzwkj2023-280);2021年国家级大学生创新创业训练项目(202110660028);2020年省级大学生创新训练项目(S202010660053);贵州省教育厅平台项目(黔教技[2022]019号)。

摘　　要：目的探讨麻疹减毒活疫苗Hu191株(MV-Hu191)对4T1乳腺癌细胞上皮-间质转化(epithelial mesenchymal transition,EMT)、增殖及迁移的影响。方法采用CCK-8法和克隆形成试验检测MV-Hu191对4T1细胞增殖的影响;细胞划痕试验检测MV-Hu191对4T1细胞迁移的影响;Western blot法检测MV-Hu191对4T1细胞EMT通路蛋白(MMP-2、MMP-9、E-cadherin)表达的影响。建立雌性BALB/c小鼠4T1荷瘤小鼠模型,将模型小鼠分为对照组(PBS)、MV-Hu191(1×106TCID50)组和紫杉醇组(15 mg/kg),每组10只,每2 d瘤内注射药物100μL,连续给药5次,28 d后,观察MV-Hu191对小鼠生存期、体内成瘤性及转移的影响;HE染色观察肺组织及肿瘤组织病理特征;免疫组化染色观察肿瘤组织EMT通路蛋白(MMP-2、MMP-9、E-cadherin)表达情况。结果体外试验结果表明,与对照组相比,MV-Hu191能抑制4T1细胞增殖及迁移(F分别为2.811和13.535,P分别为0.001和0.002),下调4T1细胞MMP-2和MMP-9表达(F分别为45.433和9.744,P分别为0.011和0.038),上调E-cadherin表达(F=7.001,P=0.032)。体内试验结果表明,与对照组相比,MV-Hu191能显著延长荷瘤小鼠生存时间(F=18.079,P=0.000),并降低肿瘤质量(F=8.301,P=0.003)和肺结节转移数量(F=33.792,P=0.000);MV-Hu191组肿瘤组织间隙较小,细胞呈圆形,肺泡轮廓清晰可见;MV-Hu191组肿瘤组织MMP-2和MMP-9表达显著降低(F分别为6.705和9.047,P分别为0.028和0.023),E-cadherin表达显著上升(F=3.468,P=0.039)。结论MV-Hu191能显著抑制4T1乳腺癌细胞增殖及迁移,拮抗4T1荷瘤小鼠成瘤性和肺转移,延长小鼠生存期,其抗乳腺癌作用的可能机制是调节EMT通路蛋白表达。Objective To investigate the effects of live attenuated measles vaccine Hu191 strain(MV-Hu191)on epithelial mesenchymal transition(EMT),proliferation and migration of 4T1 breast cancer cells.MethodsCCK-8 and clone formation assay were used to analyze the effect of MV-Hu191 on the proliferation of 4T1 cells;The effect of MV-Hu191 on 4T1cell migration was analyzed by cell scratch test;The expression of EMT pathway proteins(MMP-2,MMP-9,E-cadherin)in4T1 cells was detected by Western blot;4T1 tumor-bearing mouse model was established in female BALB/c mice.The model mice were divided into control group(PBS),MV-Hu191(1×106TCID50)group and paclitaxel group(15 mg/kg),with 10 mice in each group,and injected into tumor at the dosage of 100μL every 2 d for 5 times.At 28 d after administration,the effects of MV-Hu191 on survival time,tumorigenicity and metastasis in vivo were observed;The pathological characteristics of lung tissue and tumor tissue were observed by HE staining under microscope;The expression of EMT pathway proteins(MMP-2,MMP-9 and E-cadherin)in tumor tissue was detected by immunohistochemical staining.Results The results of in vitro experiment showed that,compared with the control group,MV-Hu191 inhibited the proliferation and migration of 4T1 cells(F=2.811 and 13.535,P=0.001 and 0.002,respectively),down regulated the expression of MMP-2 and MMP-9(F=45.433 and 9.744,P=0.011 and 0.038,respectively),and up regulated the expression of Ecadherin(F=7.001,P=0.032);The results of in vivo experiment showed that MV-Hu191 significantly prolonged the survival time of tumor-bearing mice,and decreased the tumor quality(F=8.301,P=0.003)and the number of pulmonary nodules metastasis compared with the control group(F=33.792,P=0.000);MV-Hu191 treated tumor tissue gap was small,the cells were round,and the alveolar contour was clearly visible;The expression of MMP-2 and MMP-9 in MVHu191 treated tumor tissue decreased significantly(F=6.705 and 9.047,P=0.028 and 0.023,respectively),while the expression of E-cadherin increased

关 键 词：麻疹减毒活疫苗Hu191 4T1乳腺癌细胞 细胞增殖 细胞迁移 上皮-间质转化 

分 类 号：R735.35[医药卫生—肿瘤]