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微小RNA-590-5p通过靶向RECK调控口腔鳞状细胞癌的细胞增殖、凋亡、迁移和侵袭。

MiR-590-5p regulates cell proliferation, apoptosis, migration and invasion in oral squamous cell carcinoma by targeting RECK.

作者信息

Bao Wei-Wei, Shi You-Ling, Ma Yan, Qu Xing-Hui, Pang Guang-Ming, Yang Lei

机构信息

Department of Orthodontics, Dongfeng Stomatological Hospital, Hubei University of Medicine, Shiyan, Hubei Province, China.

出版信息

Histol Histopathol. 2021 Mar;36(3):355-365. doi: 10.14670/HH-18-306. Epub 2021 Jan 15.

Abstract

OBJECTIVE

To discover the role of miR-590-5p in oral squamous cell carcinoma (OSCC) progression and the corresponding mechanism via the targeting RECK.

METHODS

OSCC (n=85) and normal oral tissues (n=60) were collected to quantify the miR-590-5p expression by using qRT-PCR. Then SCC-15 and OEC-M1 cells were selected and divided into Mock, inhibitor NC, miR-590-5p inhibitor, si-RECK and miR-590-5p inhibitor + si-RECK groups. Dual-luciferase reporter gene assay was used to verify if miR-590-5p could target RECK. The biological behaviors of OSCC cells were evaluated by MTT, Wound-healing, Transwell and Flow cytometry. The expression of miR-590-5p and RECK was measured by qRT-PCR and Western blotting , respectively.

RESULTS

Overexpression of miR-590-5p was found in OSCC tissues. The expression of miR-590-5p was significantly associated with the clinical TNM stage, differentiation degree, and lymph node metastasis of OSCC. RECK was identified as a direct target of miR-590-5p. Compared with the Mock group, cells in the miR-590-5p inhibitor group were decreased in terms of proliferation, invasion, and migration, and increased in cell apoptosis, accompanied by down-regulated miR-590-5p, Bcl-2/Bax and MMP-9, and up-regulated RECK. By contrast, si-RECK group presented completely opposite changes, and si-RECK reversed the inhibitory effect of miR-590-5p inhibitor on the OSCC cell growth.

CONCLUSION

MiR-590-5p expression was obviously increased in OSCC, and inhibiting miR-590-5p enhanced the expression of its target gene RECK, thereby suppressing proliferation, migration and invasion of OSCC cells and promoting apoptosis of OSCC cells.

摘要

目的

通过靶向RECK发现miR-590-5p在口腔鳞状细胞癌(OSCC)进展中的作用及相应机制。

方法

收集85例OSCC组织和60例正常口腔组织,采用qRT-PCR定量miR-590-5p表达。然后选取SCC-15和OEC-M1细胞,分为Mock、抑制剂NC、miR-590-5p抑制剂、si-RECK和miR-590-5p抑制剂+si-RECK组。采用双荧光素酶报告基因检测法验证miR-590-5p是否可靶向RECK。通过MTT、伤口愈合实验、Transwell实验和流式细胞术评估OSCC细胞的生物学行为。分别采用qRT-PCR和蛋白质免疫印迹法检测miR-590-5p和RECK的表达。

结果

在OSCC组织中发现miR-590-5p过表达。miR-590-5p的表达与OSCC的临床TNM分期、分化程度及淋巴结转移显著相关。RECK被确定为miR-590-5p的直接靶点。与Mock组相比,miR-590-5p抑制剂组细胞的增殖、侵袭和迁移能力降低,细胞凋亡增加,同时miR-590-5p、Bcl-2/Bax和MMP-9表达下调,RECK表达上调。相比之下,si-RECK组呈现完全相反的变化,且si-RECK逆转了miR-590-5p抑制剂对OSCC细胞生长的抑制作用。

结论

OSCC中miR-590-5p表达明显升高,抑制miR-590-5p可增强其靶基因RECK的表达,从而抑制OSCC细胞的增殖、迁移和侵袭,促进OSCC细胞凋亡。

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