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

doi:10.16098/j.issn.05290-1356.2020.03.012

摘要/Abstract

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。

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

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.

Key words: MicroRNA, Esophageal squamous cell, Proliferation, Invasion, Radiosensitivity, Flow cytometry

中图分类号:

R735.1

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.

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.

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