乳腺癌CAF中IL-6介导的上皮间质转化及其在肿瘤三苯氧胺耐受中的作用  

Epithelial-mesenchymal transition of breast cancer CAF mediated by IL-6 and its role in tumor tamoxifen resistance

作　　者：孙龙[1] 李旭军[1] SUN Long;LI Xu-Jun(The Second Hospital of Ningbo , Ningbo , Zhejiang 315010, China)

机构地区：[1]宁波市第二医院,浙江宁波315010

出　　处：《中国妇幼保健》2018年第9期2109-2112,共4页Maternal and Child Health Care of China

基　　金：宁波市自然科学基金(2013A610223)

摘　　要：目的探讨乳腺癌相关的成纤维细胞(CAF)中白细胞介素-6(IL-6)介导的上皮间质转化及其在肿瘤三苯氧胺耐受中的作用。方法选择2015年6月-2017年7月宁波市第二医院收治的乳腺癌患者50例,设为乳腺癌组;选择同期治疗的乳腺纤维瘤患者45例,设为乳腺纤维瘤组,收集乳腺癌、乳腺纤维瘤组织,采用流式细胞筛选技术完成上述不同标本成纤维细胞的筛选,并进行体外稳定生长、传代;采用趋化因子芯片分析比较乳腺癌及其他成纤维细胞分泌趋化因子表达水平;采用乳腺癌CAF细胞转染针对IL-6基因的siRNA载体质粒,分析IL-6分泌情况;采用免疫荧光显微镜及Real-time PCR、Western blot检测EMT相关指标,在CAF下进行共同培养,Real-time PCR、Western blot检测PI3K/Akt/mTOR及IL-6/JAK/STAT3信号通路。结果倒置显微镜下结果表明:两种细胞经过CAF中IL-6介导后乳腺癌细胞连接相对疏松,多边形态消失,部分细胞变为梭形,并且存在伪足;而在乳腺腺瘤中CAF中IL-6介导的细胞变化不明显;乳腺癌CAF分泌细胞因子IL-6表达水平,高于乳腺腺瘤组织(P<0.05);乳腺癌CAF分泌细胞因子IL-6表达水平,均高于癌旁组织与对照组(P<0.05);细胞划痕试验结果表明:CAF处理后乳腺癌细胞迁移距离为(18.54±3.05)mm,CAFsiRNA处理后乳腺癌细胞迁移距离为(23.50±3.28)mm,CAF、CAFsiRNA及IL-6联合处理后乳腺癌细胞迁移距离为(29.30±4.07)mm;Transwell小室侵袭试验结果表明:CAF处理后乳腺癌细胞侵袭数量为(43.45±3.60)个、CAFsiRNA处理后乳腺癌细胞侵袭数量为(68.95±4.50)个、CAF、CAFsiRNA及IL-6联合处理后乳腺癌细胞侵袭数量为(85.63±5.77)个;Western blot检测结果表明:CAF、CAFsiRNA-IL-6及IL-6处理后PI3K/Akt/mTOR及IL-6/JAK/STAT3信号表达,高于单一CAF、CAFsiRNA-IL-6(P<0.05)。结论乳腺癌CAF中IL-6介导的上皮间质转化能诱导EMT的发生,能刺进肿瘤的增殖和侵袭转移,在肿瘤三苯氧胺耐受中发�Objective To explore epithelial-mesenchymal transition of breast cancer fibroblast(CAF） mediated by interleukin-6(IL-6） and its role in tumor tamoxifen resistance. Methods From June 2015 to July 2017,50 breast cancer patients treated in the Second Hospital of Ningbo were selected as breast cancer group,and 45 breast fibroadenoma patients treated in the hospital were selected as breast fibroadenoma group. Breast cancer tissue samples and breast fibroadenoma tissue samples were collected,respectively,then fibroblast cells were screened by flow cytometry,in vitro stable growth and passage was carried out. Chemokine chip analysis was used to compare the expression levels of chemokines secreted by fibroblast cells in breast cancer group and breast fibroadenoma group. Breast cancer CAF cells were used to transfected into siRNA vector targeting to IL-6 gene to analyze the secretion of IL-6. Immunofluorescence microscopy,real-time PCR,and Western blot were used to detect the related indexes of epithelial-mesenchymal transition,CAF cells were cocultured. Real-time PCR and Western blot were used to detect PI3 K/Akt/m TOR and IL-6/JAK/STAT3 signal pathways. Results The results of inverted microscope showed that the junction of breast cancer cells mediated by IL-6 was loose in the two groups,polygon morphology disappeared,partial cells changed to shuttle shape,pseudopodium was observed. In breast fibroadenoma cells,the change of cells mediated by IL-6 was not significant. The expression level of IL-6 in breast cancer tissue was significantly higher than those in breast fibroadenoma tissue,para-carcinoma tissue,and control group(P〈0. 05）. Cell scratch assay showed that the migration distance of breast cancer cells after CAF treatment was(18. 54±3. 05） mm,and the migration distance of breast cancer cells after CAFsiRNA treatment was(23. 50± 3. 28） mm,the migration distance of breast cancer cells after CAF,CAFsiRNA,and IL-6 treatment was(29. 30 ± 4. 07） mm. Transwell chamber invasion assay showed that the numbe

关 键 词：乳腺癌 成纤维细胞 白细胞介素-6 上皮间质转化 三苯氧胺 细胞转染针 

分 类 号：R737.3[医药卫生—肿瘤]