miR-133b 通过靶向 CDCA8 抑制肺腺癌细胞的增殖、迁移和侵袭。

miR-133b inhibits cell proliferation, migration, and invasion of lung adenocarcinoma by targeting CDCA8.

机构信息

Department of Cancer Radiotherapy and Chemotherapy, Zhejiang Quhua Hospital, Quzhou, 324004, China.

Department of Oncology, Zhejiang Quhua Hospital, Quzhou, 324004, China.

出版信息

Pathol Res Pract. 2021 Jul;223:153459. doi: 10.1016/j.prp.2021.153459. Epub 2021 Apr 28.

Abstract

OBJECTIVE

Lung adenocarcinoma (LUAD) is the most common type of lung cancer. This study aims to explore the mechanism by which CDCA8 regulates cell proliferation, invasion, and migration of LUAD, and to generate novel insights into targeted therapy of LUAD.

METHODS

Expression profiles of mature microRNAs (miRNAs) and mRNAs, along with clinical data of LUAD were downloaded from TCGA database for differential analysis and survival analysis to mine differentially expressed mRNAs. qRT-PCR was used to detect the expression of CDCA8 and miR-133b in LUAD cell lines, and western blot was used to detect protein expression. The effects of CDCA8 on the proliferation, migration, and invasion of LUAD cells were detected by CCK-8 assay, scratch healing assay, and Transwell assay. Bioinformatics predicted the target miRNA of CDCA8, and dual-luciferase reporter gene assay was used to verify the binding relationship between miR-133b and CDCA8.

RESULTS

Data from TCGA-LUAD showed that CDCA8 was significantly overexpressed in LUAD tissue, while its upstream miRNA (miR-133b) was significantly lowly expressed. The result of dual-luciferase test showed that miR-133b targeted CDCA8. The results of in vitro functional experiments showed that overexpression of CDCA8 could promote the proliferation, invasion, and migration of LUAD cells, and miR-133b could reverse this promotion by targeting CDCA8.

CONCLUSION

This study found that CDCA8 was a carcinogenic factor in LUAD cells and it was regulated by upstream miR-133b. miR-133b could inhibit proliferation, invasion, and migration of LUAD cells by targeting CDCA8.

摘要

目的

肺腺癌(LUAD)是最常见的肺癌类型。本研究旨在探讨 CDCA8 调节 LUAD 细胞增殖、侵袭和迁移的机制,为 LUAD 的靶向治疗提供新的思路。

方法

从 TCGA 数据库中下载 LUAD 的成熟 microRNA(miRNA)和 mRNA 的表达谱及临床数据,进行差异分析和生存分析,挖掘差异表达的 mRNAs。qRT-PCR 检测 LUAD 细胞系中 CDCA8 和 miR-133b 的表达,Western blot 检测蛋白表达。CCK-8 assay、划痕愈合assay 和 Transwell assay 检测 CDCA8 对 LUAD 细胞增殖、迁移和侵袭的影响。生物信息学预测 CDCA8 的靶 miRNA,双荧光素酶报告基因assay 验证 miR-133b 与 CDCA8 的结合关系。

结果

TCGA-LUAD 数据显示,CDCA8 在 LUAD 组织中明显过表达,而其上游 miRNA(miR-133b)明显低表达。双荧光素酶试验结果表明,miR-133b 靶向 CDCA8。体外功能实验结果表明,CDCA8 的过表达可促进 LUAD 细胞的增殖、侵袭和迁移,而 miR-133b 可通过靶向 CDCA8 逆转这种促进作用。

结论

本研究发现 CDCA8 是 LUAD 细胞中的致癌因子,受上游 miR-133b 调控。miR-133b 可通过靶向 CDCA8 抑制 LUAD 细胞的增殖、侵袭和迁移。

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