天津医药

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食管癌细胞系中Nanog的表达对顺铂的影响

石磊芝   

  1. 辽宁医学院 研究生院
  • 收稿日期:2011-09-19 修回日期:2011-11-08 出版日期:2012-02-15 发布日期:2012-02-15
  • 通讯作者: 石磊芝

Nanog Expressed in Esophageal Cancer Cell-line and Sensitized to Cisplatin

Lei-Zhi SHI   

  • Received:2011-09-19 Revised:2011-11-08 Published:2012-02-15 Online:2012-02-15
  • Contact: Lei-Zhi SHI

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摘要： ［摘要］背景与目的：食管癌化疗耐药十分常见，癌干细胞理论认为这是由于食管癌中存在食管癌干细胞，对传统的化疗具有抗药性。本研究是检测干细胞基因Nanog在食管癌细胞系TE-1中的表达再应用RNAi下调其表达对顺铂敏感性的影响。 方法：通过细胞免疫荧光检测食管癌细胞系TE-1中Nanog蛋白的表达；应用LipofectAMINE 2000将带荧光染料的Nanog siRNA转染进细胞中，通过荧光显微镜观察转染效率，再通过RT-PCR和Western Blot检测siRNA的沉默效率；应用MTT进行细胞活性检测，转染后24小时加入顺铂，继续孵育48小时后测得不同转染组的 IC50浓度，后应用顺铂5μg /ml测得不同转染组的生存率。 结果：Nanog蛋白在食管癌细胞系TE-1中高表达，以核表达为主；细胞转染率为(67.57±16.90)％，Nanog siRNA沉默效率可高达85%以上，转染12小时后加入顺铂48小时后Nanog siRNA组IC50（4.03μg/ml），与阴性对照组（5.17μg/ml）和空白组（5.26μg/ml）相比均显著降低并有统计学差异（P<0.05）。结论：Nanog在食管癌细胞系TE-1中高表达，并且降低Nanog的表达后能够提高食管癌细胞对化疗药物顺铂的敏感性。

关键词: 食管癌, Nanog, 顺铂, 耐化疗药, siRNA

Abstract: ［Abstract］ Background and purpose：Esophageal squamous cell carcinomas (ESCC) chemotherapy drug resistance is very common, the cancer stem cells theory suggested that this was due to existing the esophageal cancer stem cells which is resistant to traditional chemotherapy. This study is testing stem cell gene Nanog expression in ESCC cell lines TE-1 and its Downregulation by RNAi Sensitizes cells to Cisplatin. Methods：In this study, we detected the expression of Nanog in the ESCC cell lines TE-1 by immunofluorescence. Then we used fluorescence labeled small interfering RNA (siRNA) to block Nanog expression while evaluating the effect of Nanog siRNA by RT-PCR and western blot , and the combined effects with Cisplatin in ESCC cell line TE-1. Results：Nanog is Highly Expressed in ESCC Cell-line TE-1and the main expression in the nucleus.The Nanog expression at the protein and mRNA level in Nanog siRNA-transfected group was reduced by 85% as compared with that in the negative control group. The cells exposed to Nanog siRNA in the presence of Cisplatin showed a significant decrease in IC50 （4.03μg /ml）compared with control siRNA (5.17μg /ml)and mock transfection (5.26μg /ml), (p < 0.05). Conclusion：The results showed that Nanog is highly expressed in ESCC cell-line TE-1 and Nanog siRNA efficiently decreased Nanog expression . Treatment with Nanog siRNA in combination with cisplatin enhanced chemosensitivity.

Key words: Esophageal squamous cell carcinomas, Nanog, Cisplatin, resistance to therapy, siRNA