长链非编码RNA LINC00885在人食管癌细胞中的作用研究

doi:10.19586/j.2095-2341.2021.0195

生物技术进展 ›› 2022, Vol. 12 ›› Issue (3): 419-426.DOI: 10.19586/j.2095-2341.2021.0195

长链非编码RNA LINC00885在人食管癌细胞中的作用研究

薛姗(), 唐铎, 赵子杰, 洪启浩, 王谦, 刘紫佳, 周志祥()

北京工业大学环境与生命学部，北京 100124

收稿日期:2021-12-16 接受日期:2022-01-06 出版日期:2022-05-25 发布日期:2022-05-26
通讯作者: 周志祥
作者简介:薛姗 E-mail：1228132199@qq.com；
基金资助:
国家自然科学基金项目(42077399)

Study on the Role of Long Non⁃coding RNA LINC00885 in Human Esophageal Cancer Cells

Shan XUE(), Duo TANG, Zijie ZHAO, Qihao HONG, Qian WANG, Zijia LIU, Zhixiang ZHOU()

Department of Environment and Life Sciences，Beijing University of Technology，Beijing 100124，China

Received:2021-12-16 Accepted:2022-01-06 Online:2022-05-25 Published:2022-05-26
Contact: Zhixiang ZHOU

摘要/Abstract

摘要：

探讨长链非编码RNA LINC00885对人食管癌细胞EC109增殖、迁移与侵袭的影响。构建LINC00885基因过表达质粒（pcDNA3.1-LINC00885）和shRNA敲低质粒（pLKO.1-LINC00885）。分别采用集落形成实验检测EC109细胞增殖能力；划痕实验检测EC109细胞横向迁移能力；Transwell实验检测EC109细胞纵向迁移能力及侵袭能力；流式实验检测LINC00885对于细胞周期的调控；实时定量PCR方法检测LINC00885 mRNA转录水平；Western blot检测BMP7及上皮间充质转化（epithelial-to-mesenchymal transition，EMT）通路相关蛋白（VIMENTIN、β-catenim和ZO-1）表达水平。在过表达LINC00885的EC109细胞中，细胞的增殖能力、迁移能力及侵袭能力均显著增强，BMP7蛋白表达升高；而在敲低表达LINC00885的EC109细胞中，细胞的增殖能力、迁移能力与侵袭能力均显著降低，BMP7蛋白表达也随之降低。另外，在过表达LINC00885的EC109细胞中，EMT通路蛋白VIMENTIN、β-catenim表达水平显著升高，而ZO-1表达水平显著降低。通过探究LINC00885对食管癌细胞增殖、迁移能力的影响，旨在验证LINC00885在食管癌中的功能，以期为临床治疗食管癌奠定理论基础。

关键词: 长链非编码RNA, 食管癌, LINC00885, EC109细胞, 增殖迁移

Abstract:

To investigate the effect of long non-coding RNA LINC00885 on the proliferation， migration and invasion of human esophageal cancer cell EC109 in this study. The LINC00885 gene overexpression plasmid （pcDNA3.1-LINC00885） and shRNA knockdown plasmid （pLKO.1-LINC00885） were constructed. Colony formation experiment was used to detect the proliferation ability of EC109 cells； the scratch healing experiment was used to detect the lateral migration ability of EC109 cells； the Transwell chamber experiment was used to detect the longitudinal migration ability and invasion ability of EC109 cells； the flow cytometry experiment was used to detect the regulation of LINC00885 on the cell cycle； real-time quantitative PCR method was used to detect the LINC00885 mRNA transcription level； Western blot was used to detect the expression levels of BMP7 and epithelial-to-mesenchymal transition （EMT） pathway related proteins （VIMENTIN， β-catenim and ZO-1）. In EC109 cells overexpressing LINC00885， the proliferation， migration and invasion capabilities of the cells were significantly enhanced， and the expression of BMP7 protein was increased. While LINC00885 down-expression significantly inhibited the proliferation， migration and invasion capabilities of EC109 cells， and BMP7 protein expression was decreased. In addition， overexpressing LINC00885 in EC109 cells， the expression levels of EMT pathway proteins VIMENTIN and β-catenim increased significantly， while the expression levels of ZO-1 decreased significantly. By exploring the effect of LINC00885 on the proliferation and migration of esophageal cancer cells， this paper aimed to verify the function of Linc00885 in esophageal cancer， in order to lay a theoretical foundation for clinical treatment of esophageal cancer.

Key words: long non-coding RNA, esophageal cancer, LINC00885, EC109, proliferation and migration

中图分类号:

R735.1

薛姗, 唐铎, 赵子杰, 洪启浩, 王谦, 刘紫佳, 周志祥. 长链非编码RNA LINC00885在人食管癌细胞中的作用研究[J]. 生物技术进展, 2022, 12(3): 419-426.

Shan XUE, Duo TANG, Zijie ZHAO, Qihao HONG, Qian WANG, Zijia LIU, Zhixiang ZHOU. Study on the Role of Long Non⁃coding RNA LINC00885 in Human Esophageal Cancer Cells[J]. Current Biotechnology, 2022, 12(3): 419-426.

图/表 4

图1 生物信息学及PT?qPCR验证LINC00885在食管癌中的表达A：RNA-seq检测LINC00885在食管癌及邻近组织中的表达情况；B：LINC00885在临床食管癌患者中的生存曲线；C：LINC00885在食管癌细胞早期SHEE26代和晚期SHEE79代的表达水平。***表示与SHEE26代相比，差异P<0.001水平有统计学意义。

Fig.1 Bioinformatics and PT?qPCR verification of LINC00885 expression in esophageal cancer

图2 LINC00885对食管癌细胞增殖及细胞周期的影响A：过表达LINC00885在EC109细胞中的表达量；B：敲低LINC00885在EC109细胞的表达量；C：使用pcDNA3.1?LINC00885和pLKO.1?LINC00885转染EC109细胞后的克隆形成测定；D：使用pcDNA3.1?LINC00885和pLKO.1?LINC00885转染EC109细胞后的Transwell测定；E：划痕实验检测过表达LINC00885对EC109细胞迁移的影响；F：划痕实验检测敲低LINC00885对EC109细胞迁移的影响；G：流式细胞仪检测过表达LINC00885对EC109细胞周期的影响；H：流式细胞仪检测敲低LINC00885对EC109细胞周期的影响。*、**、***分别表示与空载体对照相比，差异在P<0.05、P<0.01、P<0.001水平有统计学意义。

Fig.2 The effect of LINC00885 on the proliferation and cell cycle of esophageal cancer cells

图3 LINC00885对下游蛋白BMP7表达的影响A：生物信息学分析预测LINC00885与下游蛋白BMP7的相关性；B：Western blot检测LINC00885过表达对下游蛋白BMP7表达的影响；C：Western blot检测LINC00885低表达对下游蛋白BMP7表达的影响。*表示与空载体对照相比，差异在P<0.05水平有统计学意义。

Fig.3 The effects of LINC00885 on the expression of downstream protein BMP7

图4 LINC00885调节食管癌中EMT通路相关蛋白表达情况注：*表示与空载体对照相比，差异在P<0.05水平有统计学意义。

Fig.4 The situation of LINC00885 regulates EMT pathway-related protein expression in esophageal cancer

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长链非编码RNA LINC00885在人食管癌细胞中的作用研究

Study on the Role of Long Non⁃coding RNA LINC00885 in Human Esophageal Cancer Cells

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