microRNA-107通过靶向调节<i>CACNA2D1</i>抑制喉癌细胞的增殖和侵袭能力

黄朝平; 王轶; 黄石; 王振晓; 刘良发

doi:10.13201/j.issn.2096-7993.2020.10.011

microRNA-107通过靶向调节CACNA2D1抑制喉癌细胞的增殖和侵袭能力

- 1.
  成都医学院第一附属医院耳鼻咽喉头颈外科(成都，610500)
- 2.
  首都医科大学附属北京友谊医院耳鼻咽喉头颈外科
基金项目:
成都医学院科研课题(No：CYZ17-33)

详细信息

通讯作者: 刘良发，E-mail：liuliangfa301@263.net

中图分类号: R739.65

收稿日期: 2019-12-31

刊出日期: 2020-10-05

MicroRNA-107 inhibits the proliferation and invasion of laryngeal squamous cell carcinoma cells by targeting CACNA2D1

- 1.
  Department of Otolaryngology Head and Neck Surgery, the First Affiliated Hospital of Chengdu Medical College, Chengdu, 610500, China
- 2.
  Department of Otolaryngology Head and Neck Surgery, Beijing Friendship Hospital, Capital Medical University

More Information

Corresponding author: LIU Liangfa, E-mail: liuliangfa301@263.net

Received Date: 31 December 2019

Publish Date: 05 October 2020

摘要

摘要: 目的检测microRNA-107(miR-107)和钙通道蛋白基因CACNA2D1在喉癌组织中的表达, 探讨miR-107与CACNA2D1的靶向关系, 分析miR-107对喉癌细胞的增殖、侵袭及克隆形成能力的影响。方法收集40例喉癌及癌旁正常组织标本, 运用qRT-PCR检测miR-107和CACNA2D1的表达, 使用免疫印迹实验检测2种组织中钙通道蛋白α2δ1的表达; 使用双荧光素酶报告基因检测miR-107对CACNA2D1的调节作用; 在人喉癌细胞TU212及TU686中过表达或敲减miR-107, 检测喉癌细胞增殖、克隆形成及侵袭能力的变化。结果 miR-107在喉癌组织中的表达显著低于癌旁正常组织, 而CACNA2D1的表达则正好相反, 差异有统计学意义(P < 0.05);喉癌组织中的α2δ1表达明显高于癌旁正常组织(P < 0.05);双荧光素酶报告基因实验证实, miR-107通过与CACNA2D1基因mRNA的3'-UTR端2个位点(202-209, 902-908)靶向结合, 从而抑制CACNA2D1的表达及其生物学作用; 细胞实验显示, 过表达miR-107后喉癌细胞增殖、克隆形成及侵袭能力明显下降(P < 0.05), 而敲减miR-107后细胞增殖、克隆形成和侵袭能力明显增强(P < 0.05)。结论 miR-107通过靶向调节CACNA2D1从而抑制喉癌细胞的增殖、克隆形成及侵袭能力。
- microRNA-107 /
- CACNA2D1 /
- 喉肿瘤 /
- 增殖 /
- 侵袭
Abstract: Objective To detect the expression of microRNA-107(miR-107) and the calcium channel protein gene CACNA2D1 in laryngeal cancer tissues, to investigate the targeting relationship between miR-107 and CACNA2D1, and to analyze the effects of miR-107 on the proliferation, invasion and colony forming ability of laryngeal cancer cells.Method Laryngeal cancer tissues and normal adjacent tissue samples from 40 patients with laryngeal cancer were collected, and qRT-PCR was used to detect the expression of miR-107 and CACNA2D1; Western Blot assay to detect the expression of α2δ1 in the above two tissues; the dual-luciferase reporter gene was used to detect the regulatory effect of miR-107 on CACNA2D1; after overexpression or knockdown of miR-107 in human laryngeal cancer cells TU212 and TU686, changes in the proliferation, clone formation, and invasion ability of laryngeal cancer cells were detected.Result The expression of miR-107 in laryngeal cancer tissues was significantly lower than that in adjacent normal tissues, while the expression of CACNA2D1 was just the opposite, the difference was statistically significant (P < 0.05); the expression of α2δ1 in laryngeal cancer tissues is significantly higher than in normal tissues(P < 0.05); dual-luciferase reporter experiments confirmed that miR-107 binds to the 3'-UTR (202-209, 902-908) of the CACNA2D1 mRNA, thereby inhibiting the expression of CACNA2D1 and its biological effects; cell experiments showed that the proliferation, invasion, and clone formation of laryngeal cancer cells were significantly reduced after miR-107 overexpression (P < 0.05), and the cell proliferation, clone formation, and invasion were significantly enhanced after miR-107 was knocked down (P < 0.05).Conclusion miR-107 inhibits the proliferation, clone formation, and invasion of laryngeal cancer cells by targeting CACNA2D1.
- microRNA-107 /
- CACNA2D1 /
- laryngeal neoplasms /
- proliferation /
- invasiveness

HTML全文

图 1 CACNA2D1和miR-107在2种组织中的相对表达水平比较

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图 2 WB检测LSCC患者喉癌组织与癌旁正常组织中α2δ1蛋白相对表达情况

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图 3 qRT-PCR检测结果

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图 4 双荧光素酶报告基因检测

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图 5 miR-107抑制LSCC细胞的增殖率

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图 6 2种LSCC细胞的克隆形成实验结果

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图 7 5组喉癌细胞间侵袭能力比较

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表 1 各基因引物

基因	正向	反向
Has-miR-107	5’-TGCGCAGCAGCATTGTACAGGGC-3’	5’-CCAGTGCAGGGTCCGAGGTATT-3’
U6	5’-CGCTTCGGCAGCACATATAC-3’	5’-AAATATGGAACGCTTCACGA-3’
CACNA2D1	5’-GCATTGGAAGCGGAGAAAGT-3’	5’-GGAATATGGACTGCTGCGTG-3’
GAPDH	5’-TCAAGAAGGTGGTGAAGCAGG-3’	5’-CAAAGGTGGAGGAGTGGGT-3’

下载: 导出CSV

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