微小RNA-21转染介导食管鳞癌细胞生物学行为及细胞放射敏感性的变化

邵志雄 谢俊杰 徐振华 曾程 宋青玲 易峰涛

解剖学报 ›› 2020, Vol. 51 ›› Issue (3) : 385-391.

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解剖学报 ›› 2020, Vol. 51 ›› Issue (3) : 385-391. DOI: 10.16098/j.issn.05290-1356.2020.03.012
肿瘤生物学

微小RNA-21转染介导食管鳞癌细胞生物学行为及细胞放射敏感性的变化

  • 邵志雄 谢俊杰 徐振华 曾程 宋青玲 易峰涛*
作者信息 +

 Transfection of microRNA-21 mediating the biological behavior and cellular radiosensitivity of esophageal squamous carcinoma cells

  • SHAO Zhi-xiong XIE Jun-jie XU Zhen-hua ZENG Cheng SONG Qing-ling YI Feng-tao*
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文章历史 +

摘要

目的 探讨微小RNA-21(miRNA-21)表达介导食管鳞癌细胞的生物学特性。  方法  转染miRNA-21正义表达载体、miRNA-21空载体及miRNA-21抑制剂,通过Real-time PCR实验,以食管鳞状上皮癌TE-1正常细胞为对照组,检测转染结果。采用细胞计数试剂盒-8(CCK-8)对细胞增殖活性进行检测;采用流式细胞术检测细胞凋亡能力;通过Transwell小室和划痕实验观察TE-1细胞侵袭能力及迁移能力;通过集落形成实验观察TE-1细胞对放疗敏感性变化;采用SPSS 19.0统计学软件对所得数据进行分析。  结果  Real-time PCR结果显示,转染miRNA-21抑制剂后,TE-1细胞中miRNA-21的表达水平明显降低(P<0.05)。相比正常细胞生长组,阳性对照组、阴性对照组转染miRNA-21抑制剂后,TE-1细胞增殖能力降低,细胞凋亡加快(P<0.05)。此外,实验抑制组中细胞侵袭和迁移能力下降。集落形成实验分析显示,抑制miRNA-21表达明显提高了TE-1细胞对放射的敏感性。  结论  下调miRNA-21可降低食管鳞状上皮癌TE-1细胞的增殖、侵袭、迁移能力及放疗的敏感性,是未来治疗食管癌的潜在靶点。

Abstract

Objective  To study the biological properties of microRNA(miRNA)-21 expression in esophageal squamous cell carcinoma.  
 Methods  MiRNA-21 sense expression vector, miRNA-21 empty vector and miRNA-21 inhibitor were transfected and detected by fluorescence Real-time PCR. The normal cells of esophageal squamous cell carcinoma TE-1 were used as the control group, and the transfection result  were detected. The cell proliferation activity was detected by cell counting kit-8(CCK-8), and the apoptosis ability was detected by flow cytometry. The invasion ability and migration ability of TE-1 cells were observed by Transwell chamber and scratch test. Changes in the sensitivity of TE-1 cells to radiotherapy were observed by colony formation experiments. The data were analyzed using SPSS 19.0 software.   Results  The result  of Real-time PCR showed that the expression level of miRNA-21 in TE-1 cells was significantly decreased after transfected with miRNA-21 inhibitor (P<0.05). Compared with the normal cell growth group, positive control group and negative control group, the proliferation of TE-1 cells was reduced and apoptosis was accelerated after transfection with miRNA-21 inhibitor (P<0.05). In addition, the ability of cell invasion and migration in the experimental inhibition group was decreased. Clone formation assays showed that inhibition of miRNA-21 expression significantly increased the sensitivity of TE-1 cells to radiation.   Conclusion  Down-regulation of miRNA-21 reduces the proliferation, invasion, migration and sensitivity of radiotherapy in esophageal squamous cell carcinoma TE-1 cells, and is a potential target for the future treatment of esophageal cancer.

关键词

微小RNA / 食管鳞癌细胞 / 增殖 / 侵袭 / 放射敏感性 / 流式细胞术

Key words

MicroRNA / Esophageal squamous cell / Proliferation / Invasion / Radiosensitivity / Flow cytometry

引用本文

导出引用
邵志雄 谢俊杰 徐振华 曾程 宋青玲 易峰涛. 微小RNA-21转染介导食管鳞癌细胞生物学行为及细胞放射敏感性的变化[J]. 解剖学报. 2020, 51(3): 385-391 https://doi.org/10.16098/j.issn.05290-1356.2020.03.012
SHAO Zhi-xiong XIE Jun-jie XU Zhen-hua ZENG Cheng SONG Qing-ling YI Feng-tao.  Transfection of microRNA-21 mediating the biological behavior and cellular radiosensitivity of esophageal squamous carcinoma cells[J]. Acta Anatomica Sinica. 2020, 51(3): 385-391 https://doi.org/10.16098/j.issn.05290-1356.2020.03.012
中图分类号: R735.1    

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