付志璇,朱玉萍,罗 奇.癌相关成纤维细胞离体培养中激活性状的研究[J].肿瘤学杂志,2015,21(3):195-200.
癌相关成纤维细胞离体培养中激活性状的研究
The Study on the Activated State of Cancer-associated Fibroblasts Cultured in vitro
投稿时间:2014-09-09  
DOI:10.11735/j.issn.1671-170X.2015.03.B007
中文关键词:  乳腺肿瘤  癌相关成纤维细胞  平滑肌肌动蛋白
英文关键词:breast neoplasms  cancer-associated fibroblasts  smooth muscle actin(SMA)
基金项目:国家自然科学基金(81372210);浙江省医药卫生科技计划(2011KYB023)
作者单位
付志璇 浙江省肿瘤医院 
朱玉萍 浙江省肿瘤医院 
罗 奇 浙江省肿瘤医院 
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中文摘要:
      摘 要:[目的] 研究离体培养的癌相关成纤维细胞(CAFs)能否保持激活性状。[方法] 免疫组织化学和免疫荧光法检测乳腺癌及正常乳腺组织成纤维细胞激活标志物平滑肌肌动蛋白(α-SMA)表达情况。利用免疫细胞化学检测体外培养CAFs及正常乳腺组织来源成纤维细胞内α-SMA表达状态;Western blot检测CAFs和正常乳腺组织来源的成纤维细胞的α-SMA和PDGFR代次表达情况。[结果] 乳腺癌组织内CAFs呈α-SMA广泛表达。然而免疫荧光检测示只有部分CAFs呈α-SMA阳性表达。体外培养的CAFs免疫细胞化学检测显示只有部分细胞呈α-SMA阳性表达。Western blot检测显示一定代次后CAFs仍能保持PDGFR和α-SMA的表达。[结论] CAFs是异质性细胞群;CAFs体外有限传代后仍能维持激活性状。
英文摘要:
      Abstract:[Purpose] To investigate whether the activated state of cancer-associated fibroblasts (CAFs) cultured in vitro could be kept.[Methods] Fibroblasts activated marker α-SMA in breast cancer and normal tissues were detected by immunohistochemical staining (IHC) and immunofluorescent assay(IF). α-SMA in primary cultured CAFs and fibroblasts derived from normal breast tissue(NF) were detected by immunocellulerchemistry (ICC). α-SMA and PDGFR in CAFs and NF at different passages were detected by Western blot.[Results]α-SMA was widely expressed in CAFs in breast cancer tissue. However,IF showed merely partial fibroblasts in cancer mesenchymal tissue expressing α-SMA. ICC performed in cultured CAFs also demonstrated partial cells expressing α-SMA. Western blot assay for α-SMA and PDGFR showed CAFs were able to keep the both activated markers after 6 passages. [Conclusion] CAFs have the heterogeneity of cells. And CAFs are able to keep their activated state after several passages in vitro.
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