Abstract

Objective

This study aims to explore the mechanism of miR-30d-5p in regulating the development of lung squamous cell carcinoma (LUSC) via targeting DBF4.

Methods

Bioinformatics methods were employed to analyze the differentially expressed genes in LUSC tissue microarray. qRT-PCR was employed to detect the expression of miR-30d-5p and DBF4 mRNA in normal human bronchial epithelial cells and LUSC cells. CCK-8 was used to detect LUSC cell activity. Wound healing assay was employed to detect the migratory ability of LUSC cells. Transwell was employed to detect invasive ability. Dual-luciferase reporter assay was used to detect the targeting relationship between miR-30d-5p and DBF4. Western blot was used to detect the protein expression of marker molecules associated with epithelial–mesenchymal transition (EMT).

Results

In this study, the expression of miR-30d-5p in LUSC cell lines was found to be obviously low compared with that in normal human bronchial epithelial cell line, which was opposite to the expression of DBF4. Dual-luciferase reporter assay verified that miR-30d-5p could target DBF4 and the overexpression of miR-30d-5p downregulated the expression of DBF4. Overexpression of DBF4 promoted the proliferation, migration, invasion, and EMT of LUSC, whereas over-expression of miR-30d-5p could weaken the promotion of DBF4 on cancer cells.

Conclusion

miR-30d-5p downregulates the expression of DBF4 to regulate the development of LUSC.

中文翻译：

---

miR-30d-5p 通过靶向 DBF4 抑制肺鳞状细胞癌的增殖、迁移和侵袭

摘要

客观的

本研究旨在探讨miR-30d-5p通过靶向DBF4调控肺鳞状细胞癌（LUSC）发展的机制。

方法

采用生物信息学方法分析LUSC组织微阵列中的差异表达基因。采用qRT-PCR检测正常人支气管上皮细胞和LUSC细胞中miR-30d-5p和DBF4 mRNA的表达。CCK-8 用于检测 LUSC 细胞活性。伤口愈合试验用于检测 LUSC 细胞的迁移能力。Transwell 用于检测侵袭能力。双荧光素酶报告基因检测用于检测 miR-30d-5p 和DBF4之间的靶向关系。Western印迹用于检测与上皮 - 间质转化（EMT）相关的标记分子的蛋白质表达。

结果

本研究发现 miR-30d-5p 在 LUSC 细胞系中的表达明显低于正常人支气管上皮细胞系中的表达，这与DBF4的表达相反。双荧光素酶报告基因检测证实 miR-30d-5p 可以靶向DBF4并且 miR-30d-5p 的过表达下调DBF4的表达。DBF4的过表达促进了LUSC的增殖、迁移、侵袭和EMT，而miR-30d-5p的过表达可以削弱DBF4对癌细胞的促进作用。

结论

miR-30d-5p 下调DBF4的表达以调节 LUSC 的发展。