解剖学报 ›› 2018, Vol. 49 ›› Issue (4): 455-460.doi: 10.16098/j.issn.0529-1356.2018.04.007

• 肿瘤生物学 • 上一篇    下一篇

TRIM28siRNA通过活化E2F转录因子1增加非小细胞肺癌PAa细胞对依托泊苷的敏感性

刘镭1 高亚贤1 王建平1 赵学荣1 许倩2* 胡旺3 刘美奇4   

  1. 1.承德医学院基础医学院免疫学教研室; 2.承德医学院基础医学院基础医学研究所; 3.承德医学院基础医学院2014级临床本科9班; 4.承德医学院基础医学院2014级麻醉本科1班,河北 承德067000
  • 收稿日期:2017-08-14 修回日期:2017-11-22 出版日期:2018-08-06 发布日期:2018-08-06
  • 通讯作者: 许倩 E-mail:xuqian@163.com
  • 基金资助:
    河北省自然科学基金资助项目;河北省人才工程培养经费资助科研项目

TRIM28siRNA enhancing the sensitivity of non-small cell lung cancer cell PAa to etoposide through activating E2F transcription factor 1

LIU Lei1 GAO Ya-xian1 WANG Jian-ping1 ZHAO Xue-rong1 XU Qian 2* HU Wang3 LIU Mei-qi4   

  1. 1.Department of Immunology; 2.Basic Medicine Laboratory; 3.Class 9 Grade 2014, Clinical Undergraduate; 4. Class 1 Grade 2014, Anesthesia Undergraduate, Institute of Basic Medicine, Chengde Medical College, Hebei Chengde067000, China
  • Received:2017-08-14 Revised:2017-11-22 Online:2018-08-06 Published:2018-08-06
  • Contact: XU Qian E-mail:xuqian@163.com

摘要:

目的  探讨siRNA沉默TRIM28(tripartite motif containing 28)增强非小细胞肺癌(NSCLC)PAa细胞对依托泊苷的敏感性和可能的机制。方法  应用RNA干扰技术沉默TRIM28基因在PAa细胞中的表达;TRIM28siRNA单独或联合依托泊苷处理PAa细胞后,应用MTT法和集落形成实验检测各组细胞的增殖;流式细胞术检测细胞凋亡;Western blotting方法检测各组细胞中E2F转录因子1(E2F1)的表达水平。结果  在PAa中沉默了TRIM28的表达;TRIM28siRNA联合依托泊苷显著抑制PAa细胞的增殖和集落形成,明显诱导细胞凋亡;联合处理的PAa细胞中E2F1mRNA和蛋白水平的表达明显增加,同时抗凋亡蛋白Bcl-2表达减少。结论TRIM28siRNA联合化疗药物依托泊苷对抑制NSCLC细胞生长有明显效果。

关键词: 非小细胞肺癌, PAa细胞, TRIM28, RNA干扰, 依托泊苷, 免疫印迹法,

Abstract:

Objective  To investigate the effect of small interfering RNA (siRNA)-mediated silencing of TRIM28(tripartite motif containing 28)on the sensitivity of non-small cell lung cancer cell (NSCLC) PAa to etoposide and explore the possible mechanism. Methods  The expression of TRIM28 in PAa cell was silenced by RNA interference. MTT and colony formation assay were used to assess the inhibitory effect of TRIM28siRNA alone or in combination with etoposide on the growth and proliferation of PAa cells. The apoptosis of different groups was detected by flow cytometry(FCM ) and the expression of E2F transcription factor 1(E2F1) was evaluated by Western blotting. Results  A PAa cell line silenced with the TRIM28siRNA was established. TRIM28 in combination with etoposide significantly inhibited the proliferation and colony formation of PAa cells. TRIM28siRNA in combination with etoposide induced apoptosis and increased the expression of E2F1 at mRNA and protein levels. Conclusion  TRIM28siRNA in combination with etoposide has an obvious effect on the growth of NSCLC cells, suggesting a new clinical treatment strategy for NSCLC.

Key words: Non-small cell lung cancer, PAa cell, TRIM28, RNA interference, Etoposide, Western blotting, Human