| 谢聪颖,苏华芳,景 钊.放射抗拒性食管癌细胞株KYSE-150R上皮—间质转化特性鉴定[J].肿瘤学杂志,2012,18(7):504-507. |
| 放射抗拒性食管癌细胞株KYSE-150R上皮—间质转化特性鉴定 |
| Identification of Epithelial-mesenchymal Transition Phenotype in Radioresistant Esophageal Carcinoma Cell Line KYSE-150R |
| 投稿时间：2012-04-26 |
| DOI： |
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| 中文关键词: 放射抗拒 食管肿瘤 上皮间质转化 KYSE-150R细胞株 |
| 英文关键词:radiation resistance esophageal neoplasms epithelial-mesenchymal transitions |
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| 中文摘要: |
| 摘 要：［目的］鉴定放射抗拒性食管癌细胞株KYSE-150R获得上皮—间质转化(EMT) 表型。［方法］ 食管鳞状细胞癌细胞株KYSE-150经分次反复照射、单克隆培养，筛选出具有放射抗拒性细胞株KYSE-150R。克隆形成实验计算细胞存活率，拟合生存曲线并计算放射生物学参数。高倍显微镜下观察细胞形态学变化。采用RT-PCR、Western Blot和免疫荧光检测上皮表型标志物E钙黏蛋白(E-cad)、间质表型标志物波形蛋白(VIM)和转录因子Slug、Snail与Twist的表达。［结果］ 构建的食管癌细胞株KYSE-150R比其亲本对射线更加抗拒，经传代50代后仍保留放射抗拒性。放射抗拒的食管癌细胞株KYSE-150R发生EMT相符的形态学改变：细胞呈纺锤体状，极性消失。与母株相比，KYSE-150R中E-cad表达下降，VIM、Slug及Snail表达上调，Twist表达未见明显变化。免疫荧光染色证实细胞膜E-cad蛋白的丢失，上调的β-catenin移位至细胞核。［结论］ 放射抗拒的食管癌细胞株KYSE-150R获得了EMT表型，其与食管癌放射抗拒的形成有关。 |
| 英文摘要: |
| Abstract：［Purpose］ To identify the acquisition of epithelial-mesenchymal transition phenotype in radioresistant esophageal carcinoma cell line KYSE-150R. ［Methods］ The radioresistant cell line KYSE-150R was established by fractionated irradiation (FIR) and monoclone culture. Cell survival was accounted by clonogenic assays and the morphological changes were observed by microscopy. RT-PCR，Western Blot and immunofluorescence were performed to examine expression of E-cadherin，vimentin，Slug，Snail and Twist. ［Results］ The established esophageal carcinoma cell line KYSE-150R showed higher radioresistance than parental cell line. The KYSE-150R cell line maintained radioresistant phenotype for at least 5 months with 50 passages after the cessation of radiation. KYSE-150R cells showed phenotypic changes consistent with EMT: spindle-cell shape，and loss of polarity. Compared with mother cell line the expression of E-cadherin downregulated and the expression of vimentin，Snail and Slug upregulated in KYSE-150R cells. However，the expression of Twist did not show marked changes. ［Conclusion］ Radioresistant KYSE-150R cells acquire phenotype of EMT，and it might relate to radioresistance for esophageal carcinoma. |
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